Quantitative pollen viability analysis is a critical but labor-intensive step in plant reproductive biology. Existing deep-learning Segment Anything Models (SAM) fail to reliably segment viable pollen in Alexander-stained anthers. To address this, we fine-tuned an existing Cellpose-SAM model for pollen segmentation. We integrated it into PAT (Pollen Analysis Tool), a cross-platform desktop application. PAT features instance segmentation with interactive quality control, an in-app model retraining module, and publication-ready statistical outputs. We deployed PAT in an EMS suppressor screen of semi-sterile Arabidopsis smg7-6 mutants, enabling efficient candidate prioritization for whole-genome sequencing and mapping of the candidate mutation. This screen led to the identification of a point mutation in CAP-D2 (capd2-2), a Condensin I subunit, that rescues the smg7-6 meiotic phenotype. Notably, mutation in a Condensin II subunits (CAP-D3 and CAP-H2) does not confer rescue. Further characterization suggests the capd2-2 allele is hypomorphic, showing no defects in vegetative growth, chromocenter compaction, or transposable element silencing. Collectively, we demonstrate that accessible AI tools have the potential to bridge gaps in plant phenotyping and accelerate the pace of biological discovery.
Why it matches plant phenotyping methods花粉生存性を画像から自動推定するセグメンテーション手法とソフトウェアPATの開発が研究の中心であり、植物表現型計測ツールとして明確に該当する。
titlePAT: An Image Analysis Tool for Automated Scoring of Pollen in Alexander-Stained Anthers
Reproduction assets foundThe paper's Data availability statement explicitly deposits the authors' PAT pollen-phenotyping tool (the software implementing the paper's computational analysis, including the fine-tuned CPSAM segmentation model support) as open source on GitHub. Note: the full repository URL in the text (https://github.com/Riha-429[Code · public17
Data availability
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Pollen Analysis tool (PAT) is available as open source tool at Github repository (https://github.com/Riha-429
Lab/Pollen-Analysis-Tool).
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Figure legends
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Fig. 1. Cellpose performance on Alexander-stained anther cross-sections across varying pollen
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densities.
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Representative cross-sections of Alexander-stained anthers showing a range of pollen densities, from
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low (top rows, light staining) to high (bottom rows, dense reOpen asset ↗Pollen-Analysis-Toolpdf-raw-page:17 lines:1-64Code / dataset availability confirmedOpenAlex · Crossref · checked 14 Sept 2026
The automation and standardization of hyperspectral imaging, particularly at high spatial resolution, are essential for advancing plant phenomics and supporting diverse plant science research. We developed HyperBird, a hyperspectral microscopic imaging robot, to automate the acquisition of up to 351 leaf-disc samples in a single tray within approximately 2.4 h and thereby support scalable plant experiments. System calibration established a spatial resolution of 24.3 μ m (full width at half maximum; FWHM), a spectral resolution of 1.78 nm (FWHM), and a depth of field of 4.53 mm, producing hyperspectral data cubes of 2195 × 2000 × 950 pixels across the 400–1000 nm spectral range. System-level and biological-sample consistency assessments confirmed stable spectral measurements during extended scanning sessions and across independently prepared trays. Thermal evaluation confirmed that the 150 W illumination design introduced minimal sample heating during scanning. HyperBird was first evaluated using grape downy and powdery mildews, where no consistent measurable effect of repeated imaging on pathosystem development was detected under the tested experimental conditions. Subsequently, HyperBird was used to characterize spatiotemporal spectral progression in grapevine leaves inoculated with Plasmopara viticola . An automated regional spectral tracing pipeline was developed to isolate spectra from retrospectively traced regions and compare them with whole-leaf averaged spectra across 0–9 days post-inoculation (DPI). Categorical mixed-effects modeling showed that these spatially resolved regional spectra exhibited significant disease-associated spectral change by 5 DPI, with a sharp transition at 6 DPI, whereas whole-leaf spectra showed delayed and weaker disease-aligned changes beginning at 7 DPI. These results demonstrate that HyperBird enables high-throughput, spatially resolved hyperspectral imaging for quantifying localized plant disease progression and provides a scalable platform for studying spectral biology across plant phenotyping applications.
Why it matches plant phenotyping methodsHyperBirdの高スループット・ハイパースペクトル画像取得ロボットを開発し、校正、再現性、加熱影響、病害進展の定量性能を検証しており、植物フェノタイピング手法が中心である。
abstractWe developed HyperBird, a hyperspectral microscopic imaging robot, to automate the acquisition of up to 351 leaf-disc samples in a single tray within approximately 2.4 h and thereby support scalable plant experiments.
Reproduction assets foundThe paper's Data Availability statement points to a public GitHub repository containing the authors' code and processed data supporting the phenotyping analyses; raw hyperspectral image data are request-only.Code · publicData Availability
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The code and processed data supporting the findings of this study are available in the GitHub
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repository at https://github.com/jy773Cornell/HyperBird-Robot. Raw hyperspectral image
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data are available from the corresponding author upon reasonable request due to file size and storage
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constraints.
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Supplementary Materials
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Supplementary materials accompany this article as a separate document (supplementary.pdf).
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Supplementary Figure S1. Representative GSAM-based segOpen asset ↗HyperBird-Robotpdf-raw-page:39 lines:1-75Code / dataset availability confirmedEurope PMC · checked 5 Sept 2026
Stomatal traits are key microscopic phenotypes for evaluating plant physiology, stress responses, and crop breeding potential. However, in vivo high-magnification microscopy often suffers from a shallow depth of field, causing noticeable defocus blur across different spatial locations and making it difficult to capture clear and complete stomatal structures in a single image. Multi-focus image fusion offers a practical solution, yet existing methods typically rely on supervised training, paired data, or hand-crafted rules, limiting their use in real agricultural microscopy scenarios. In this study, we propose an unsupervised multi-focus fusion framework for reconstructing fully focused stomatal microscopic images. The method integrates two-dimensional feature extraction with three-dimensional cross-focal-plane modeling to capture both spatial details and complementary information across focal planes. A max-response-guided spatial gating module is introduced to enhance focused regions while suppressing defocused responses. Additionally, dual sharpness priors based on perceptual features and wavelet high-frequency information enable pixel-wise pseudo-supervised learning without requiring all-in-focus ground-truth images. The model also predicts a probabilistic focal-plane volume for interpretable all-in-focus reconstruction. Experiments on a maize multi-focus image dataset demonstrate that the proposed method achieves superior or competitive performance across multiple fusion metrics, with entropy (EN), edge information preservation ( Q AB∕F ), Chen-Blum contrast metric ( Q CB ), and visual information fidelity for fusion (VIFF) reaching 7.43, 0.21, 0.41, and 1.01, respectively. Ablation studies confirm the effectiveness of the 3D modeling, spatial gating, and dual-prior sharpness supervision. More importantly, when the fused images serve as input to a YOLO-based stomatal instance segmentation model, the proposed method yields the best segmentation accuracy, with mAP50 and mAP50-95 reaching 0.9937 and 0.9121, respectively. Phenotypic measurements derived from the segmentation masks show high consistency with manual annotations, with the highest coefficient of determination R 2 = 0.97 achieved for stomatal count. These results indicate that the framework can act as an effective front-end module for automated microscopic stomatal phenotyping in agriculture.
Why it matches plant phenotyping methods植物の気孔表現型を対象に、マルチフォーカス画像融合、セグメンテーション、形質測定までを中核的に開発・検証しているため。
abstractwe propose an unsupervised multi-focus fusion framework for reconstructing fully focused stomatal microscopic images.
Reproduction assets foundThe authors state their data and code are publicly available on GitHub, covering the multi-focus stomatal microscopy dataset and the UMF-stomata fusion/phenotyping code.Code · publicOur data and code are available at: https://github.com/Longer-S/UMF-Stomata.Open asset ↗Longer-S/UMF-Stomatahtml-lines:640-655Dataset · publicOur data and code are available at: https://github.com/Longer-S/UMF-Stomata.Open asset ↗Longer-S/UMF-Stomatahtml-lines:683-756Code / dataset availability confirmedEurope PMC · checked 5 Sept 2026
Abstract Cross-sectional analysis is considered the reference method for measuring cotton fiber fineness and maturity; however, its widespread use has been limited by labor-intensive sample preparation and manual image analysis. The objective of this study was to develop and validate a reproducible deep-learning-based workflow for automated segmentation and quantitative analysis of cotton fiber cross-sections. A total of 249 composite light microscopy images of cotton fiber cross-sections were collected and manually annotated to generate training and validation datasets. A YOLO11m instance segmentation model was developed to identify cotton fiber and lumen regions and automatically extract quantitative traits, including fiber area, lumen area, fiber perimeter, and lumen perimeter. The workflow integrates automated image segmentation, post-processing, quantitative trait extraction, and data export to facilitate reproducible cotton fiber phenotyping. Model performance was evaluated using mean Average Precision (mAP), and workflow outputs were validated against Adobe Photoshop using descriptive comparisons of six cross-sectional traits. The model achieved Box mAP50 scores of 0.984 for cotton fiber regions and 0.789 for lumen regions, demonstrating high segmentation accuracy. The automated workflow substantially reduced manual analysis time while producing measurements with central tendencies comparable to those obtained using Adobe Photoshop. To facilitate reproducibility and adoption, the workflow, trained model weights, and supporting documentation are publicly available through GitHub and a Hugging Face web application. The workflow substantially increases analytical throughput while providing a reproducible and publicly accessible method for automated cotton fiber cross-sectional phenotyping, facilitating quantitative analysis for cotton genetics and breeding research.
Why it matches plant phenotyping methods綿繊維横断面の画像分割、形質抽出、検証を目的とした再現可能な深層学習ワークフローの開発であり、植物フェノタイピング手法が研究の中心である。
abstractThe objective of this study was to develop and validate a reproducible deep-learning-based workflow for automated segmentation and quantitative analysis of cotton fiber cross-sections.
Reproduction assets foundThe paper's cotton fiber cross-section phenotyping workflow (YOLO11m segmentation pipeline, trained model weights, example images/outputs) is explicitly stated as publicly available via a GitHub repository and a Hugging Face web application, with URLs matching the allowed list.Code · publicThe complete source code, training scripts, dataset configuration, example input images, example outputs,
and supporting documentation are publicly available through the GitHub repository:
https://github.com/RifeLab/cotton-lumen-microOpen asset ↗RifeLab/cotton-lumen-micropdf-page:11 lines:1-47Code · publicThe cotton fiber image analysis workflow is publicly available through a web-based application hosted on
Hugging Face at: https://huggingface.co/spaces/chaneylc/cotton_fiber_microscopy_measureOpen asset ↗pdf-page:11 lines:1-47Code / dataset availability confirmedEurope PMC · checked 5 Sept 2026
Idesia polycarpa Maxim. is a premier woody oil species in Guizhou Province, China, whose fruit yield and oil quality largely depend on effective pollination and fertilization. However, limited research on pollen viability and germination has hindered industrial progress. To address this gap, a comprehensive evaluation framework for elite I. polycarpa germplasm was developed, integrating micromorphological analysis, optimized staining protocols, and in vitro germination assay. Scanning electron microscopy (SEM) revealed that I. polycarpa pollen, while genetically conserved at the genus level-characterized by prolate shapes, tricolporate apertures, and reticulate exine ornamentation-exhibits notable micromorphological variation among genotypes. Of the nine staining protocols tested (2,3,5-triphenyl tetrazolium chloride [TTC], carbol fuchsin, acetocarmine, methylene blue, Alexander, peroxidase, 2,5-diphenylmonotetrazolium bromide [MTT], I2-KI, and red ink), TTC and red ink were the most effective, offering clear chromatic distinction between viable and non-viable pollen. Through orthogonal experimental designs, genotype-specific optimal media for in vitro germination were identified: 0.40 g/L H3BO3, 0.01 g/L KNO3, 0.02 g/L Ca(NO3)2·4H2O, and 0.20 g/L KH2PO4 for STZ-6; and 0.20 g/L H3BO3, 0.02 g/L KNO3, 0.02 g/L Ca(NO3)2·4H2O, and 0.10 g/L KH2PO4 for STZ-9. Regression analysis confirmed a highly significant positive correlation (P < 0.01) between in vitro germination rates and the staining results from both TTC and red ink across various concentrations. Notably, 5% TTC and 30% red ink exhibited the highest coefficients of determination. A hierarchical evaluation strategy is thus proposed: the 5% TTC method is recommended for precise laboratory quantification due to its stability, while the 30% red ink method, due to its ease of use, is suited for rapid field-based screening. This study provides valuable insights into the morphological characteristics of I. polycarpa pollen and establishes a standardized evaluation framework, supporting germplasm innovation and optimizing pollination management.
Why it matches plant phenotyping methods花粉の生存性・発芽という植物の生殖形質を対象に、染色法とin vitro発芽法を最適化・検証し、標準化した評価フレームワークを開発しているため、方法論が中心である。
abstracta comprehensive evaluation framework for elite I. polycarpa germplasm was developed, integrating micromorphological analysis, optimized staining protocols, and in vitro germination assay.
Reproduction assets foundThe article's Data Availability statement points to a public Biostudies deposit containing the study's data (pollen morphology measurements, staining viability counts, and in vitro germination results). No author analysis code or trained models are mentioned.Dataset · publicData Availability: The data that support the findings of this study are openly available in Biostudies at https://doi.org/10.6019/S-BSST3125 .Open asset ↗Biostudies · S-BSST3125lines:176-186Code / dataset availability confirmedOpenAlex · Europe PMC · checked 5 Sept 2026
Abstract Background Stomata and pavement cells are fundamental components of the leaf epidermis, jointly regulating gas exchange, water loss, and leaf surface expansion. Stomata size, aperture, density, and pavement cell morphology are critical parameters for assessing plant transpiration efficiency, epidermal growth dynamics, and adaptive responses to environmental constraints. Despite their biological importance, quantifying stomatal and pavement-cell traits remains seldom not generalized, simple, and fast enough . Manual or semi-automated approaches limit large-scale phenotyping and restrict the integration of epidermal morphology into crop-improvement pipelines aimed at developing climate-resilient varieties with optimised stomatal patterning. To address such limitations, we developed Stomatalia , a deep learning-based platform designed to automate and standardise the quantification of stomatal and pavement cell traits. The algorithm was trained on epidermal images of cultivated and wild potato and tomato genotypes grown under optimal and abiotic-stress conditions. Stomatalia automatically detects stomata and pavement cells and extracts a broad range of morphological and integrative epidermal parameters, enabling high-throughput phenotyping within a unified workflow. Results Prior to platform development, we optimised a rapid, minimally-destructive leaf-printing protocol that generates negative impressions of the leaf surface within 40–100 s. Transparent positive prints were subsequently produced and imaged under a light microscope at scale settings ranging from 20 to 200 μm. The resulting images are analysed using Stomatalia’s user-friendly web-based interface, which runs an instance-segmentation deep learning algorithm to detect, count, and calculate stomatal and pavement cell parameters. The platform outputs structured files containing raw measurements, derived integrative traits, and associated metadata, facilitating downstream statistical and physiological analyses. Algorithm evaluation on independent datasets demonstrated high performance within the validated dicot imaging domain, with F 1 -scores ranging from 0.86 to 0.94 depending on image scale, species, and resolution, and high segmentation overlap for both stomata and pavement cells. The generality of stomatal detection was also tested on spring onion, chickpea, balsam poplar, and wheat in cross-species feasibility tests, although performance was more variable in monocots, and pavement-cell segmentation remained species- and architecture-dependent. Benchmarking against another publicly available app further showed that, under the tested web interface settings and image types, Stomatalia exhibited closer agreement with manual counts and substantially faster processing times. The practical performance of Stomatalia was further tested in a proof-of-concept trial with potato plants subjected to optimal irrigation and a long, gradual drought. The platform reliably quantified epidermal traits despite variations in leaf morphology and image quality, supporting the integrated interpretation of stomatal and pavement-cell responses under stress. Conclusions We developed Stomatalia as a robust, user-friendly deep learning platform for automated, high-throughput analysis of bright-field leaf epidermal images across varying magnifications and resolutions. Stomatalia facilitates rapid, reproducible, and coordinated phenotyping of stomatal and pavement cells by integrating methodological standardisation, computational automation, and multi-trait extraction in a single analytical workflow. Its strongest current application is the analysis of high-quality dicot leaf-print images, particularly in species and imaging conditions similar to those used for model training and validation. Cross-species and benchmark analyses further define its current scope: stomatal detection can be transferred to some additional epidermal architectures, whereas robust pavement-cell segmentation in monocots or highly divergent species will require further annotation and model retraining. Within these defined boundaries, Stomatalia provides a flexible and extensible framework for studying stomatal and pavement cell morphology and environmental plasticity, while also supporting broader efforts to dissect and optimise plant responses to abiotic stress.
Why it matches plant phenotyping methods気孔・舗装細胞の形態形質を画像から自動抽出する深層学習プラットフォームを開発し、独立データで性能評価・比較検証しているため、植物フェノタイピング手法が中心である。
abstractwe developed Stomatalia , a deep learning-based platform designed to automate and standardise the quantification of stomatal and pavement cell traits.
Reproduction assets foundThe authors publicly deposited the paper's test image datasets (raw/input leaf-print images, detection outputs, manual ground-truth counts, exported datasets) and the model file on Figshare, and provide a public Google Colab demo for running the Stomatalia algorithm. Both are paper-specific, public, and actionable.Dataset · publicThe test datasets and model file used in this work are available through the following link:
https://doi.org/10.6084/m9.figshare.32532672. The test_sets.zip archive contains the test image datasets
(cross-species and benchmark analysis), including raw/input images, detection output images, manual
ground-truth counts and exported datasets.Open asset ↗Figshare · 10.6084/m9.figshare.32532672pdf-page:25 lines:1-75Code / dataset availability confirmedEurope PMC · OpenAlex · bioRxiv · Crossref · checked 15 Sept 2026
Background Cell geometry plays a central role in determining division orientation and body axis formation during early embryogenesis in Arabidopsis thaliana . However, quantitative analysis of dynamic three-dimensional (3D) morphology remains challenging because live-imaging studies often rely on two-dimensional (2D) projections, while existing 3D reconstruction approaches, including mesh-based methods, often lose the original orientation information relative to the ovule and require labor-intensive mesh correction. In addition, embryo positional fluctuation caused by floating in liquid medium and continuous growth makes it difficult to analyze temporal morphological changes within a common coordinate system. Results We developed a robust framework for quantitative 3D and four-dimensional (4D; 3D + time) analysis of embryo initial cell (apical cell) morphology. The method first establishes a standardized 3D coordinate system by normalizing cell orientation based on the bottom plane and the optical axis of the observation. Cell morphology is then reconstructed through ellipse-based approximation of serial cross-sections extracted from stacked imaging data, enabling accurate geometric characterization without the need for complex surface mesh reconstruction. To evaluate shape anisotropy, we quantified the apical cell shape in 3D. The framework further supports the characterization of volumetric features of subsequent division, providing a basis for quantifying 3D embryogenesis. Conclusion Our framework provides a simple and noise-reduced approach for quantitative analysis of living cell morphology in 3D. We named the integrated method of combining coordinate normalization with elliptical cross-section-based reconstruction Apical3DTip. This method enables consistent comparison of cell shapes without extensive manual corrections. The method overcomes key limitations of 2D projection-based and mesh-dependent analyses and offers a practical platform for quantifying cell shape and daughter cell shapes in 3D. More broadly, it provides a quantitative foundation for exploring the relationship between cell geometry, morphodynamics, and developmental patterning in living plant embryos.
Why it matches plant phenotyping methods植物胚の細胞形態を3D・4D画像から定量化する再構成手法を開発しており、表現型取得・抽出法が研究の中心である。
abstractWe developed a robust framework for quantitative 3D and four-dimensional (4D; 3D + time) analysis of embryo initial cell (apical cell) morphology.
Reproduction assets foundThe paper's Methods availability statement explicitly deposits the Apical3DTip analysis code and associated datasets on two public GitHub repositories (main implementation and ImageJ plugin). These are paper-specific author assets for the 3D/4D apical cell reconstruction and phenotyping analysis. No separate phenotype/Code · publicctor of the fitted vertical plane:
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Because the fitted plane passes through the centroid s, the offset e was calculated as
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Availability of data and materials
The code for Apical3DTip, along with all associated datasets, is available on Github:
https://github.com/blues0910/Apical3DTip.
Apical3DTip is also available as an ImageJ plugin:
https://github.com/YusukeKimata-Moo/Apical3DTip.
Ethics approval and consent to participate
Not applicable.
Consent for publication
Not applicable.
Competing interests
The authors declare no competing interests.
Funding
This work was supported by a Japan Society for the PromotOpen asset ↗https://github.com/blues0910/Apical3DTippdf-layout-page:12 lines:1-49Code · publictroid s, the offset e was calculated as
N
MQ.
Then, the fitted plane was represented as
O G N O
MQ 0.
Availability of data and materials
The code for Apical3DTip, along with all associated datasets, is available on Github:
https://github.com/blues0910/Apical3DTip.
Apical3DTip is also available as an ImageJ plugin:
https://github.com/YusukeKimata-Moo/Apical3DTip.
Ethics approval and consent to participate
Not applicable.
Consent for publication
Not applicable.
Competing interests
The authors declare no competing interests.
Funding
This work was supported by a Japan Society for the Promotion of Science (JSPS) KAKENHI
Grant (No. JP22K15135 to H.M., JP25H01809 to Y.K., JP26K02023 tOpen asset ↗https://github.com/YusukeKimata-Moo/Apical3DTippdf-layout-page:12 lines:1-49Code / dataset availability confirmedarXiv · OpenAlex · checked 11 Sept 2026
Tomographic microscopy enables three-dimensional internal imaging but often requires expensive optical or X-ray instrumentation. Here we present an ultra-low-cost continuous-wave diffusive tomography (CWDT) system for biological samples. The system uses a smartphone microscope, a white LED coupled into an optical fiber, 3D-printed micropositioners, and a physics-based forward model optimized with machine learning. We demonstrate full-color volumetric reconstructions from a tartrazine-cleared poplar section, a scattering phantom, fungal mycelium near an Arabidopsis root, and thick poplar branch imaging with an inserted side-emitting fiber. The current results are qualitative and exploratory, but they show that scanned fiber illumination and inexpensive hardware can produce useful three-dimensional reconstruction outputs for low-cost microscopy experiments.
Why it matches plant phenotyping methods低コスト三次元断層イメージング法そのものを開発し、ポプラ組織・枝やシロイヌナズナ根近傍を対象に植物の内部構造を可視化しているため、植物形態の取得法として中心的です。
abstractHere we present an ultra-low-cost continuous-wave diffusive tomography (CWDT) system for biological samples.
Reproduction assets foundThe paper's raw imaging inputs, configurations, and reconstruction outputs for Figures 2–5 are publicly deposited on Kaggle. The analysis code repository is only 'prepared for release' (no confirmed public deposit yet), so it is listed as request-only. Hardware CAD mirrors are public but are instrument designs, not theDataset · publicFigure-level raw inputs, model configurations, selected outputs, and manifests are available through the Kaggle dataset https://www.kaggle.com/datasets/alingold/continuous-wave-diffusive-tomography .Open asset ↗continuous-wave-diffusive-tomographylines:108-129Code / dataset availability confirmedEurope PMC · checked 5 Sept 2026
Quantitative disease resistance in plants emerges from complex interactions between host tissues and pathogen growth dynamics, producing a spectrum of phenotypic responses. In plant-fungal interactions, disease is most visibly expressed through lesions that vary in number, size, shape, and color, collectively defining a lesion profile. For Cochliobolus heterostrophus, a fungus causing Southern Corn Leaf Blight of maize (Zea mays ssp. mays), we show that infection on different maize genotypes produces strikingly different lesion profiles. However, it remains unclear whether such macroscopic variation in lesion profiles corresponds to consistent differences in the three-dimensional organization of pathogen colonization within host tissue. We therefore examined variation in the three-dimensional structure of C. heterostrophus-infection networks across host genotypes representing four lesion-profile classes. Using light-sheet microscopy and filament-tracing methods adapted from neuroscience, we developed quantitative metrics to characterize infection network organization, including depth, density, shape, and spatial association with host vascular tissue. In this dataset, network depth was similar across genotypes, whereas network morphology (shape and density), spatial association with vascular bundles, hyphal segment length, and branching frequency varied. Notably, genotypes with similar quantitative resistance levels sometimes exhibited distinct patterns of fungal colonization, suggesting that comparable resistance can arise from different underlying infection dynamics. These findings indicate that lesion profiles may not uniquely predict infection network structure and highlight the utility of three-dimensional network metrics for describing variation that likely reflects multiple underlying host and pathogen processes. This multi-scale framework provides tools for linking macroscopic disease phenotypes with microscopic infection processes in quantitative disease resistance.
Why it matches plant phenotyping methods植物病斑と病原菌感染ネットワークを対象に、ライトシート顕微鏡とトレーシング法を適応し、感染構造を定量化する指標を開発・適用しており、表現型取得法が研究の中心である。
abstractUsing light-sheet microscopy and filament-tracing methods adapted from neuroscience, we developed quantitative metrics to characterize infection network organization, including depth, density, shape, and spatial association with host vascular tissue.
Reproduction assets foundThe paper's Data availability statement explicitly deposits metadata, data, and computer code as Supplementary Files accompanying the open-access publication (Supplementary Materials 1-5, including XLSX datasets and an untyped Supplementary Material 5 likely holding code). These are paper-specific phenotyping assets (eCode · publicMetadata, data, and computer code from this study are available in Supplementary Files included with the publication.Open asset ↗lines:147-204Code / dataset availability confirmedEurope PMC · checked 5 Sept 2026
PollenBB16 is an RGB pollen image dataset of Chilean flora with pixel-accurate instance segmentation masks, whose annotation was fully verified by an expert palynologist to guarantee the taxonomic reliability of every published instance. The dataset is designed to close a concrete gap in existing palynological datasets, which typically combine low taxonomic diversity, few samples per class, and low-resolution crops restricted to bounding boxes. PollenBB16 contains 16,198 brightfield optical microscopy images at the native resolution of 3088 × 2064 pixels and 36,383 pixel-accurate polygons across 16 species from the Biobío Region, spanning endemic, native and exotic species of high ecological and melliferous value such as Eucryphia glutinosa and Quillaja saponaria (endemic), Gevuina avellana and Aristotelia chilensis (native), and Medicago sativa and Brassica rapa (introduced). Each spatial position is recorded at three focal planes. The displacement along the z axis reveals features of the exine together with information on the internal structure of the grain that remain inaccessible on a single plane. From this multifocal information, more robust convolutional networks can be trained with more accurate classification. The operational quality of the dataset is backed by a leakage-safe partition that keeps the three focal planes of the same position in the same subset to avoid metric inflation, complemented by a YOLO11n-seg baseline trained for 50 epochs that reaches 0.985 mask mAP@50 on the validation set, establishing a reproducible reference point. Beyond deep learning, PollenBB16 enables interdisciplinary applications in aerobiology, biodiversity monitoring under climate change, ecological restoration of the South American temperate forest, and botanical-origin authentication of Chilean monofloral honeys.
Why it matches plant phenotyping methods植物由来の花粉粒を対象とした高解像度画像データセットとセグメンテーション基準を構築し、深層学習による画像解析を再現可能な形で検証しているため、植物フェノタイピング手法・データセットとして中心的です。
abstractPollenBB16 is an RGB pollen image dataset of Chilean flora with pixel-accurate instance segmentation masks
Reproduction assets foundThe paper's PollenBB16 pollen image dataset (16,198 multifocal RGB microscopy images with pixel-accurate instance segmentation masks) and the accompanying authors' script polygons_to_bboxes.py are publicly deposited on Zenodo (10.5281/zenodo.19830051), per the Data Availability Statement.Code · publicThe only custom code distributed with this Data Descriptor is the Python script polygons_to_bboxes.py, which regenerates the YOLO bounding-box labels in labels_bb/ from the polygon labels in labels/. The script is packaged inside the scripts/ folder of the same Zenodo repository that hosts the dataset (10.5281/zenodo.19830051) and is released under the Creative Commons Attribution 4.0 International (CC BY 4.0) license, with no restrictions on access.Open asset ↗Zenodo · 10.5281/zenodo.19830051html-lines:731-797Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
Understanding cellular growth dynamics in plants requires precise, long-term imaging of developing tissues. Cauline leaves are produced during the transition from vegetative to reproductive development and provide a useful system for studying how laminar organs diversify in form and function. While other laminar organs, such as rosette leaves and sepals, have been extensively studied, early cauline leaf development remains technically challenging to capture due to their concealed position, curved morphology, and the presence of dense trichomes. Here, we provide a complete pipeline for the dissection, confocal imaging, 2.5D segmentation, and image analysis of initiating cauline leaves in Arabidopsis thaliana . This method enables reproducible, high-resolution imaging of cauline leaves, supporting robust quantitative analysis of growth across developmental stages at cellular scale resolution. Key features • Fine dissection method for exposing initiating cauline leaves in Arabidopsis thaliana . • Long-term confocal live imaging of cauline leaf development at cellular resolution. • Optimized imaging parameters for high-fidelity 2.5D segmentation and growth analysis in MorphoGraphX.
Why it matches plant phenotyping methodsカウリン葉の成長を細胞レベルで定量化するための解剖、共焦点イメージング、2.5Dセグメンテーション、画像解析パイプラインが中心的に開発・提示されている。
abstractHere, we provide a complete pipeline for the dissection, confocal imaging, 2.5D segmentation, and image analysis of initiating cauline leaves in Arabidopsis thaliana .
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Code · public2. MorphoGraphX 2.0.1 ( https://morphographx.org/software/ ) (access date, 2026-02-26) [10–11]
3. All codes have been deposited to OSF: https://osf.io/uth78/ (access date, 2026-02-26)
Procedure
A. Plant growth
1. Sow the seeds in pots filled with moist, room-temperature soil. Add a layer of water to the bottom of the tray and cover with a lid to maintain high humidity.
Note: Space seeds sufficiently to avoid contact between the developing plants and to prevent leaf damage; typicallyOpen asset ↗OSFlines:109-143Code / dataset availability confirmedOpenAlex · Europe PMC · checked 5 Sept 2026
Accurate three-dimensional (3D) nuclear instance segmentation is a prerequisite for quantitative phenotyping in volumetric microscopy, yet remains challenging in densely packed tissues, irregular nuclear morphologies, and across heterogeneous imaging modalities. Here we present NucVerse3D, a deep-learning framework for generalized 3D nuclei instance segmentation that combines a residual attention 3D U-Net architecture with a reversible gradient-field representation for robust centroid-aware instance reconstruction. NucVerse3D is trained end to end in 3D using modality-agnostic preprocessing and isotropic scale normalization, enabling deployment across confocal microscopy, two-photon microscopy, light-sheet microscopy, micro-computed tomography, and scanning electron microscopy volumes. We benchmarked NucVerse3D on seven volumetric datasets spanning multiple species and tissues, comprising more than forty thousand manually annotated nuclei, including newly released ground-truth datasets of mouse liver tissue (control and hepatocellular carcinoma) and Drosophila brain glial nuclei. Across datasets, NucVerse3D achieved consistently high precision and competitive recall, resulting in strong F1-scores and average precision across a wide range of imaging conditions. While a modest precision-recall imbalance is observed in certain datasets, favoring high-confidence detections, this behavior reflects a conservative instance reconstruction strategy that prioritizes accurate boundary delineation and reduces false positive segmentation in densely packed and morphologically heterogeneous tissues. A single generalized model trained on pooled data matched the performance of dataset-specific models, and ablation experiments demonstrated that preprocessing and scale normalization substantially contribute to performance under strict intersection-over-union criteria. To demonstrate the biomedical utility of NucVerse3D, we applied it to 3D liver images from a mouse model of hepatocellular carcinoma (HCC) to enable spatially resolved 3D nuclear phenotyping. In healthy liver tissue, nuclear DNA content and nuclear volume exhibited a tightly regulated log-log scaling relationship. In contrast, tumor-adjacent and tumor regions displayed progressive disruption of this coupling, forming spatially coherent domains of nuclear DNA-volume decoupling that are not detectable in conventional two-dimensional histology. We quantify this phenomenon using a Nuclear Decoupling Score (NDS), revealing increased nuclear instability aligned with pathological tissue remodeling highlighting NDS as a potential quantitative biomarker of dysplastic and tumor tissue. Together, NucVerse3D provides a robust and generalizable solution for 3D nuclear instance segmentation and enables quantitative nuclear phenotyping across imaging modalities.
Why it matches plant phenotyping methods3D核セグメンテーション手法を開発し、多様な画像データセットでベンチマーク・検証したうえで、核形態とDNA量の定量的フェノタイピングに応用しており、植物対象ではないため本索引の対象外となる可能性はあるが、提示内容上はフェノタイピング手法研究として中心的である。
abstractAccurate three-dimensional (3D) nuclear instance segmentation is a prerequisite for quantitative phenotyping in volumetric microscopy
Reproduction assets foundThe paper's newly released Zenodo deposit (10.5281/zenodo.18517324) containing raw volumes, annotations, training patches, model weights, and segmentation outputs is not among the allowed URLs, so it cannot be listed. The authors' public analysis/segmentation code repository is explicitly deposited with an authors' URLCode · publicThe source code for training and predicting nuclei segmentation using NucVerse 3D is available from https://github.com/Segovia-lab/3D-Nuclei-segmentation.git .Open asset ↗https://github.com/Segovia-lab/3D-Nuclei-segmentation.gitlines:647-728Code / dataset availability confirmedEurope PMC · bioRxiv · checked 5 Sept 2026
Grapevine Trunk diseases (GTDs) represent a major threat for the wine industry. Despite several break-through, their etiology remains unclear and no curative treatment is currently available. Wood anatomy and water transport contribute to the symptoms of young plant decline. This study investigates wood anatomical alterations in two Alsatian grapevine cultivars presenting different susceptibility to GTDs, focusing on wood structure over six months of vegetative growth and in response to infection. Using a validated FasGa staining protocol, wood sections from transverse, tangential, and radial directions were stained to differentiate lignified and cellulosic tissues. Microscopic analysis was performed at x4, x10, and x40 magnifications, yielding a dataset of 4771 images. To support this high-throughput quantitative analysis of microscopy images, a computational model was developed, enabling reliable and efficient assessment of anatomical traits. Pre-established woody tissues presented higher xylem vessels diameter in Gewurztraminer than Riesling, with a dorsoventral arrangement whereas the number of vessels remained the same all over the cross section. No significant anatomical changes were observed in established woody tissues, whereas newly formed xylem anatomy showed a possible rearrangement during infection, especially in Gewurztraminer cultivar. Furthermore, colorimetric analysis quantified the lignification of woody tissues in response to wounding damage compared to un-treated plants. While definitive conclusions remain limited due to the experimental timeframe and sample variability, the findings highlight the need for longer-term studies and broader cultivar evaluation. Code and microscopy images have been made publicly available, providing a scalable digital tool for future research in plant vascular systems.
Why it matches plant phenotyping methods植物組織画像から木部解剖形質と木化を定量する計算モデルを開発・検証し、大規模画像データセットと公開コードを提供しており、表現型取得手法が研究の中心である。
abstractTo support this high-throughput quantitative analysis of microscopy images, a computational model was developed, enabling reliable and efficient assessment of anatomical traits.
Reproduction assets foundThe paper's microscopy image dataset (4771 grapevine wood images) is publicly deposited on Zenodo with an explicit DOI matching an allowed URL. The authors also state their Python analysis pipeline is available at github.com/courbot/vineside, but that URL is not among the allowed URLs, so only the Zenodo image dataset,Dataset · publicThis database can benefit the research community, and is publicly available
online at https://doi.org/10.5281/zenodo.18850060 [35].Open asset ↗Zenodo · 10.5281/zenodo.18850060pdf-page:4 lines:1-56Code / dataset availability confirmedEurope PMC · bioRxiv · Crossref · checked 5 Sept 2026
A central problem in soft and biological physics is how molecular-scale activity and remodelling coarse-grain into emergent mechanical laws at larger scales. In growing cell walls (polymeric composite materials that surround 90% of living organisms’ cells) irreversible deformation is not controlled by elastic stress alone. Instead, growth depends on the interplay between energy storage, dissipation, and the local timing of viscoelastic relaxation. Although dynamic atomic force microscopy (AFM) resolves storage and loss moduli ( E′, E″) of living walls at nanometre resolution, these observables have remained phenomenological and disconnected from constitutive field variables. Here we introduce a physics-based inversion framework that converts AFM measurements of epidermal cells of living Arabidopsis plants into spatially resolved fields of stiffness k , viscosity η , and relaxation time τ . By analysing the spatial gradients of E′ and E″, we uncover organized mechanical heterogeneities governed by cellular confinement and stress focusing. We demonstrate that the local relaxation time is encoded directly in the coupling between storage and dissipation, yielding the pointwise relation τ = (1/ ω ) ∂ E ’/∂ E ’’, where ω is the indentation frequency. This relation enables model-independent extraction of mechanical timescales and establishes a general route from nanoscale non-equilibrium rheology to continuum descriptions of growth in living and active soft materials. Significance How molecular-scale activity gives rise to tissue-scale form is a central challenge in biological physics. Although growth is fundamentally a non-equilibrium mechanical process, experimental measurements at the nanoscale have not been directly connected to the constitutive parameters that govern morphogenesis. We introduce a framework that converts dynamic atomic force microscopy maps of storage and loss moduli into spatially resolved fields of stiffness, viscosity, and relaxation time in living cell walls. By revealing that mechanical relaxation is encoded in the local coupling between elastic storage and viscous dissipation, our work provides a route from nanoscale rheology to growth-relevant mechanical timing. This establishes a quantitative bridge between molecular remodeling and continuum mechanics, enabling direct experimental constraints on multiscale theories of morphogenesis.
Why it matches plant phenotyping methods生きたArabidopsis細胞のAFM測定を物理ベースで反転し、剛性・粘性・緩和時間という植物細胞壁の機械的形質を空間的に抽出する新規フレームワークが研究の中心である。
abstractHere we introduce a physics-based inversion framework that converts AFM measurements of epidermal cells of living Arabidopsis plants into spatially resolved fields of stiffness k , viscosity η , and relaxation time τ .
Reproduction assets foundThe paper's custom AFM viscoelastic analysis code is explicitly deposited and publicly available on GitHub (ForceMetric). The underlying AFM phenotype/measurement data are only available upon request, not publicly.Code · publicAFM data were analysed in Python 3.5 using previously described routines [34] (code
available at https://github.com/jcbs/ForceMetric ).Open asset ↗jcbs/ForceMetricpdf-page:14 lines:1-56Code / dataset availability confirmedOpenAlex · Europe PMC · checked 5 Sept 2026
Peanut (Arachis hypogaea L.), a major legume crop valued for its high oil content, displays complex genotypic-phenotypic interactions shaped by environmental influences, yet these relationships remain poorly understood. We present a high-throughput phenotyping framework that captures the geometry of peanut pods using digital microscopy or smartphone imaging integrated with manifold-learning for large-scale analysis and visualization. Using over 6500 pods collected across China, we identify a geographically distinct morphological signature and demonstrate accurate cultivar discrimination. This scalable approach establishes the foundation for a Large Geometric Model capable of predicting phenotypic traits and accelerating precision agriculture. Our pipeline offers a transformative tool for peanut breeding and sustainable crop improvement.
Why it matches plant phenotyping methodsデジタル顕微鏡・スマートフォン画像と多様体学習を統合し、ピーナッツ莢の形態を大規模に取得・解析する高スループット表現型解析フレームワークが中心である。
abstractWe present a high-throughput phenotyping framework that captures the geometry of peanut pods using digital microscopy or smartphone imaging integrated with manifold-learning for large-scale analysis and visualization.
Reproduction assets foundThe authors state that the peanut pod image dataset, extracted phenotypic trait data, and the Orange Data Mining workflow (.ows) used for analysis are publicly available in their GitHub repository.Dataset · publicThe image dataset of peanut pods analyzed in this study and the extracted phenotypic trait data are publicly available in the GitHub repository: https://github.com/pengwengkung/Complex-geometry-peanut .Open asset ↗pengwengkung/Complex-geometry-peanutlines:169-192Code / dataset availability confirmedEurope PMC · bioRxiv · Crossref · checked 15 Sept 2026
Quantitative studies of plant growth and environmental responses increasingly rely on time-series imaging, yet automated segmentation remains challenging due to continuous growth, large non-rigid morphological change, and frequent self-occlusion. Traditional image-processing pipelines and task-specific deep learning models often require extensive annotated datasets and retraining, limiting portability across species, developmental stages, and imaging conditions. Here we present SAP (Segment Any Plant), a plant-focused framework that leverages the pretrained Segment Anything Model 2 (SAM2) to enable few-shot, training-free segmentation of plant time-series imagery. SAP integrates interactive prompting, automated temporal mask propagation, and centerline extraction within a web-based interface, allowing users to move from raw images to quantitative descriptors of organ shape and dynamics without programming expertise. Across multiple systems, including Arabidopsis thaliana rosette development, root growth, sunflower gravitropism, and confocal root microscopy, SAP achieves high segmentation accuracy (mean IoU 0.89–0.93) and sub-pixel centerline precision from single-frame prompting. By reducing the need for task-specific retraining, SAP provides a transferable framework for reproducible time-series phenotyping across diverse experimental contexts.
Why it matches plant phenotyping methods植物の時系列画像から器官形状・動態を抽出するセグメンテーション手法とWeb基盤を開発し、複数系で精度検証しているため、植物フェノタイピング手法が中心である。
abstractHere we present SAP (Segment Any Plant), a plant-focused framework that leverages the pretrained Segment Anything Model 2 (SAM2) to enable few-shot, training-free segmentation of plant time-series imagery.
Reproduction assets foundThe paper's authors publicly release both the SAP analysis code (GitHub repository) and the datasets generated/analyzed in the study (Zenodo), including raw images, ground-truth and SAP-generated segmentation masks, centerline validation data, and supplementary videos. Both are paper-specific, public, and directly cit.Dataset · publicCode Availability. The code is available at
https://github.com/merozlab/plant-segmentation-app.Data Availability. The datasets generated and an-
alyzed during this study are available on Zenodo at
https://doi.org/10.5281/zenodo.18732705. This includes
raw images and segmentation masks for the sunflower
gravitropism and Arabidopsis root growth experiments,
SAP-generated masks for the Lee et al. (9) and Strauss
et al. (13) datasets, centerline validation data, and supple-
mentary videos.
Funding. Y.M. acknowledges support from the Israel Sci-
ence Foundation ResOpen asset ↗zenodo · 10.5281/zenodo.18732705pdf-raw-page:9 lines:1-74Code / dataset availability confirmedOpenAlex · Europe PMC · bioRxiv · checked 5 Sept 2026
ABSTRACT Nominal hours post fertilization (hpf) are widely used to index zebrafish embryogenesis, yet under condition shifts—such as temperature change, genetic perturbation, or environmental stress—nominal time can decouple from true developmental progression. In such settings, biologically meaningful variation is better described as a systematic change in developmental tempo rather than a simple temporal offset. Here we introduce an embryo-resolved framework that treats developmental tempo as the primary quantity of interest in brightfield time-lapse imaging. We present EmbryoTempoFormer (ETF), a clip-based CNN–Transformer that predicts developmental progression from short time-lapse clips and is trained with a within-embryo temporal-difference consistency regularizer to promote temporally coherent trajectories. Crucially, we couple model predictions with an embryo-level inference and statistical workflow: temporally correlated clip-level outputs are aggregated into interpretable embryo-level tempo and stability readouts, and cross-condition effects are quantified using embryo-bootstrap confidence intervals with embryos—rather than frames or clips—as independent units, avoiding pseudo-replication. Using temperature perturbation as a representative domain shift, we robustly quantify condition-induced changes in global developmental dynamics and show that developmental delay predominantly manifests as reduced developmental tempo. This framework enables statistically principled, high-throughput phenotyping for perturbation screens, drug assays, and environmental stress studies. HIGHLIGHTS Clip-based CNN–Transformer predicts developmental time from brightfield time-lapse microscopy. Within-embryo temporal-difference consistency improves trajectory self-consistency. Embryo-level anchored tempo slopes enable interpretable cross-condition comparisons. Reproducible pipeline via code, scripts, and a Zenodo bundle with embryo-level inference Graphical abstract
Why it matches plant phenotyping methodsゼブラフィッシュ胚の発生進行・テンポをタイムラプス画像から推定するCNN–Transformerと、胚単位の統計的推定ワークフローを開発しており、表現型取得・抽出手法が中心である。ただし植物ではなく動物対象のため、この植物フェノタイピング索引では除外すべき内容。
abstractHere we introduce an embryo-resolved framework that treats developmental tempo as the primary quantity of interest in brightfield time-lapse imaging.
Reproduction assets foundThe paper analyzes public zebrafish brightfield time-lapse data (BioImage Archive S-BIAD531) and provides a public GitHub code repository plus a Zenodo reproducibility bundle containing processed arrays, model checkpoints, dataset splits, and checksums. All three are paper-specific, public, and actionable.Code · publicCode repository: https://github.com/LijiayuDeng/s-biad531-embryo-tempoformerOpen asset ↗https://github.com/LijiayuDeng/s-biad531-embryo-tempoformerpdf-page:25 lines:1-52Code / dataset availability confirmedCrossref · checked 5 Sept 2026
Abstract Background Apoplastic pH is a central regulator of plant growth, development, and environmental adaptation, influencing cell expansion, nutrient uptake, and extracellular signaling. Many studies have successfully used HPTS to monitor relative changes in apoplastic pH in plants. At the same time, research increasingly targets pH-dependent biochemical and biophysical processes. Many enzymatic activities, ion binding events, and receptor–ligand interactions depend on defined proton concentrations. Accordingly, the development of reliable approaches to measure absolute pH in living tissues is gaining importance. Methods A calibration-based workflow was developed to enable quantitative assessment of absolute apoplastic pH using ratiometric HPTS imaging. The approach integrates a simplified two-point normalization strategy with an in-vitro derived sigmoidal calibration model, thereby minimizing the need for extensive in-vivo calibration curves. Confocal imaging was performed using HPTS excited at two wavelengths followed by ratiometric image processing. Data analysis is supported by a custom Fiji plugin, Ratio2pH, which converts ratiometric images into pixel-resolved maps of absolute pH. Results In vitro characterization revealed a robust, non-linear relationship between normalized HPTS ratios and pH, enabling accurate pH estimation within the physiologically relevant range of pH 5.0–7.0. When applied in-vivo to Arabidopsis thaliana roots, the workflow yielded extracellular pH estimates consistent with the pH of the incubation medium and detected reproducible pH shifts in response to pharmacological treatments. Conclusions This workflow enables reproducible, spatially resolved measurement of absolute apoplastic pH in living plant tissues. By combining a simplified calibration strategy with accessible image analysis tools, it facilitates quantitative extracellular pH measurements and their integration into biochemical and biophysical analyses.
Why it matches plant phenotyping methods生きた植物組織の絶対アポプラストpHを画像から定量する校正ワークフローを開発・検証し、Fijiプラグインも提供しているため、植物状態の取得法が中心である。
abstractA calibration-based workflow was developed to enable quantitative assessment of absolute apoplastic pH using ratiometric HPTS imaging.
Reproduction assets foundThe paper deposits its authors' analysis code and data publicly: the Ratio2pH Fiji plugin (Zenodo 10.5281/zenodo.15599805), a Python script for sigmoidal calibration curve fitting (Zenodo 10.5281/zenodo.17303477), and source data files and raw confocal images (Freidata 10.60493/t29wb-7my86). The Zenodo 15658668 ratiom�Code · publicThe Python Script for generating a user-defined sigmoidal calibration curve is available at Zenodo: https://doi.org/10.5281/zenodo.17303477Open asset ↗Zenodo · 10.5281/zenodo.17303477lines:175-235Dataset · publicSource data files and raw images are uploaded at Freidata, the data server of the University of Freiburg, available under https://doi.org/10.60493/t29wb-7my86Open asset ↗Freidata · 10.60493/t29wb-7my86lines:175-235Code / dataset availability confirmedEurope PMC · checked 5 Sept 2026
Plants defend against pathogens such as fungi by initiating coordinated structural and chemical responses. Pathogen perception triggers rapid cytosolic calcium influx and calcium oscillations that drive defense gene expression, yet the mechanisms by which these signals encode stressor intensity and propagate systematically remain unclear. Here, we present a microfluidic system to characterize intracellular calcium dynamics in protonemal colonies of the moss Physcomitrium patens (Hedw.) upon precise and reversible exposure to fungal chitin oligosaccharides. Epifluorescent imaging of cells expressing the calcium indicator GCaMP6f revealed a rapid, coordinated calcium response to chitin addition, followed by stereotyped oscillations that subsided quickly upon stimulus removal. We implemented an unbiased image segmentation algorithm using pixel-based k -means clustering to automatically locate regions with specific oscillatory signatures. Calcium dynamics were distinct across adjacent cells, distinguishable by cell type, and significantly modulated by circadian rhythm, adaptation time within the device, and stimulus timing. Cytosolic calcium oscillations, which rose and fell symmetrically within about 60 s, occurred spontaneously during the subjective night and following short adaptation periods. Chitin elicited strong oscillations with increased frequency, amplitude, and duration, and repeated pulses entrained regular, colony-wide oscillations at the stimulation interval. This study complements prior investigations of whole plant and growth tip dynamics and provides a quantitative framework to study calcium signaling in plants, including mechanisms of signal propagation and the role of oscillation frequency on gene expression.
Why it matches plant phenotyping methods植物細胞のカルシウム動態を定量するマイクロ流体・蛍光イメージング系と自動画像セグメンテーションを開発し、植物の生理状態を抽出する方法が研究の中心である。
abstractHere, we present a microfluidic system to characterize intracellular calcium dynamics in protonemal colonies of the moss Physcomitrium patens
Reproduction assets foundThe paper's Data Availability Statement explicitly makes analysis scripts and sample data publicly available on the authors' GitHub repository (albrechtLab/moss_calcium), and the MDPI supplementary materials (plants-15-00582-s001.zip) contain the paper's timelapse calcium-imaging videos and supplementary figures. Raw/全Code · publicData are available upon request. Analysis scripts and sample data are publicly available at https://github.com/albrechtLab/moss_calcium (accessed on 1 January 2026).Open asset ↗albrechtLab/moss_calciumlines:188-251Code / dataset availability confirmedEurope PMC · bioRxiv · Crossref · checked 5 Sept 2026
Specialized host-microbe interfaces are central to cellular interactions in plants. Intracellular structures such as haustoria formed by filamentous pathogens mediate nutrient exchange and effector delivery to host cells. Despite their biological importance, the lack of quantitative frameworks has largely confined the study of these interfaces to qualitative observations, limiting our ability to compare infection strategies, cellular responses, and spatial organization across cells and tissues. Here, we present HFinder , a deep learning-based framework for automated detection, segmentation, and quantitative analysis of plant-microbe interfaces in confocal images. Using an object-centric deep learning approach, HFinder enables robust identification of haustoria, microbial hyphae, and host organelles across diverse imaging conditions and pathosystems. We demonstrate that this framework supports quantitative analyses of subcellular processes at host-microbe interfaces, including effector secretion, perturbation of host cellular processes, and immune receptor accumulation at haustoria. HFinder provides a practical and scalable solution for the systematic digitalization of plant infection imaging data and establishes a general framework for quantitative studies of cellular dynamics at host-microbe contact zones.
Why it matches plant phenotyping methods植物と微生物の界面を共焦点画像から自動検出・分割し、ハウストリア等を定量解析する深層学習手法が中心であり、植物感染状態の画像ベース表現型解析に該当する。
abstractwe present HFinder , a deep learning-based framework for automated detection, segmentation, and quantitative analysis of plant-microbe interfaces in confocal images.
Reproduction assets foundThe paper's HFinder pre-trained models (trained phenotyping models/checkpoints) are explicitly deposited on Zenodo with a public DOI matching an allowed URL. The training image dataset is also stated to be publicly available on Zenodo, but no separate authors' URL for it is given in the supplied blocks, so only the preModel / weights · publicFor
convenience, HFinder is distributed with pre-trained models that can be applied directly to confocal image
analysis (available on Zenodo: https://doi.org/10.5281/zenodo.17091805)Open asset ↗Zenodo · 10.5281/zenodo.17091805pdf-page:5 lines:1-47Code / dataset availability confirmedCrossref · Europe PMC · checked 5 Sept 2026
Understanding how plants perceive and respond to environmental and developmental cues requires tools capable of monitoring molecular signals in vivo, across whole tissues, and in real time. Genetically encoded fluorescent indicators, coupled with fluorescence microscopy, have transformed plant biology, but their application remains largely confined to small model organisms and specialized microscopy instrumentation. Here, we present MAcro Plant Projection Imaging (MAPPI), an open-source, low-cost, and modular fluorescence imaging platform for soil-grown plants beyond the model organism or seedling stage. MAPPI enables wide field-of-view, dual-projection imaging of fluorescent reporters, supporting real-time visualization of systemic signals under near-physiological conditions. We validate MAPPI by tracking calcium and l -glutamate dynamics in adult Nicotiana benthamiana plants, revealing developmentally regulated long-distance calcium waves triggered by wounding, burning, or submergence, including bidirectional shoot-to-root and root-to-shoot signaling. By democratizing access to whole-plant functional imaging, MAPPI provides a scalable tool for dissecting signal propagation, stress adaptation, and systemic communication in both model and nonmodel species.
Why it matches plant phenotyping methods植物全体の蛍光シグナルをリアルタイム取得する低コスト・オープンな画像プラットフォームを開発し、成体植物で検証しているため、植物表現型取得法が研究の中心です。
abstractHere, we present MAcro Plant Projection Imaging (MAPPI), an open-source, low-cost, and modular fluorescence imaging platform for soil-grown plants beyond the model organism or seedling stage.
Reproduction assets foundThe authors publicly deposit raw imaging data and MAPPI analysis code on Zenodo, host the MAPPI acquisition/analysis code on GitHub, and release the napari-roi-registration image registration plugin on GitHub. All are paper-specific, public, and actionable.Dataset · publicThe raw data for the images presented in the manuscript and the code to run the MAPPI system are available on Zenodo ( https://doi.org/10.5281/zenodo.15845576 ).Open asset ↗Zenodo · 10.5281/zenodo.15845576lines:170-466Code · publicThe code to run the MAPPI system is also available on the dedicated GitHub repository ( https://github.com/micropolimi/MAPPI ) along with the code used to analyze the data.Open asset ↗GitHub · micropolimi/MAPPIlines:170-466Code · publicThe software is open-source and available on GitHub ( https://github.com/GiorgiaTortora/napari-roi-registration ) and the napari-hub ( www.napari-hub.org/plugins/napari-roi-registration ).Open asset ↗GitHub · GiorgiaTortora/napari-roi-registrationlines:156-169Code / dataset availability confirmedEurope PMC · checked 5 Sept 2026
Background The plant vacuole arises by orchestrated interplay of membrane trafficking, cytoskeletal rearrangements and a variety of signaling pathways. In the root, the characteristic large central vacuole develops by endomembrane reorganization occurring mainly in the transition zone. The vacuole's bounding membrane-the tonoplast-can be visualized in vivo using fluorescent protein markers, allowing for quantitative analysis of confocal microscopy images. Tonoplast organization can thus serve as a sensitive indicator of changes to any of the processes involved in vacuole biogenesis. The Vacuolar Morphology Index (VMI) is widely accepted as a quantitative measure of vacuole structure. However, this metric has two drawbacks-it only reflects the size of the largest vacuolar compartment (missing therefore possible differences in the organization of smaller compartments), and its determination is labor intensive, limiting its use on large datasets. Results We developed an alternative metric for describing vacuole organization, named the Tonoplast Topology Index (TTI), which overcomes the above-mentioned shortcomings of the VMI. We compared the performance of our protocol with VMI on a simulated dataset and on real data. To validate the methods´ performance, we used it to confirm the previously reported differences in vacuole shape and size between Arabidopsis thaliana roots grown on the surface of an agar medium compared to those embedded inside the agar. Both VMI and TTI could efficiently detect the relatively subtle changes in vacuole organization depending on the position of the root in the agar, and provided correlated results. However, only TTI produced data with close to normal value distribution, simplifying subsequent statistical evaluation. Conclusions We present the protocol for TTI determination as a two-stage semi-automated procedure involving microscopic image analysis employing an ImageJ macro and subsequent processing of numeric data in the Jupyter Notebook environment, together with benchmarking image data. Since this implementation is freeware-based, platform-independent and (relatively) user-friendly, we hope it will find its use as a high throughput, added value alternative to the VMI metric.
Why it matches plant phenotyping methods植物液胞構造を定量化する新規指標と半自動画像解析プロトコルを開発し、シミュレーションおよび実画像で既存指標と比較・検証しているため、植物フェノタイピング手法が中心である。
abstractWe developed an alternative metric for describing vacuole organization, named the Tonoplast Topology Index (TTI)
Reproduction assets foundThe paper deposits its benchmark confocal image dataset in the EMBL-EBI BioImage Archive (S-BIAD2226) and its TTI analysis software (ImageJ macro and Jupyter/Python scripts) on GitHub, both with explicit public availability statements.Dataset · publicImage data generated and analyzed in the current study are available in the EMBL-EBI BioImage Archive repository, accession number S-BIAD2226Open asset ↗EMBL-EBI BioImage Archive · S-BIAD2226lines:141-163Code · publicArchive copy, additional sample data and possible future updates of the software tool generated here are also available at [ https://github.com/GeorgeCaldarescu/TTI-Tonoplast-Topology-Index ] .Open asset ↗GitHub · GeorgeCaldarescu/TTI-Tonoplast-Topology-Indexlines:141-163Code / dataset availability confirmedEurope PMC · OpenAlex · Crossref · checked 15 Sept 2026
Background Root hairs play a key role in plant nutrient and water uptake. Historically, root hair traits have largely been quantified manually. As such, this process has been laborious and low-throughput. However, given their importance for plant health and development, high-throughput quantification of root hair morphology could help underpin rapid advances in the genetic understanding of these traits. With recent increases in the accessibility and availability of artificial intelligence (AI) and machine learning techniques, the development of tools to automate plant phenotyping processes has been greatly accelerated. Results We present pyRootHair, a high-throughput, AI-powered software application to automate root hair trait extraction from microscope images of plant roots grown on agar plates. pyRootHair is capable of batch processing over 600 images per hour without manual input from the end user. In this study, we deploy pyRootHair on a panel of 24 diverse wheat (Triticum aestivum and Triticum turgidum ssp. durum) cultivars and uncover a large, previously unresolved amount of variation in many root hair traits. We show that the overall root hair profile falls under 2 distinct shape categories and that different root hair traits often correlate with each other. We also demonstrate that pyRootHair can be deployed on a range of plant species, including oat (Avena sativa), rice (Oryza sativa), teff (Eragrostis tef), and tomato (Solanum lycopersicum). Conclusions The application of pyRootHair enables users to rapidly screen a large number of plant germplasm resources for variation in root hair morphology, supporting high-resolution measurements and high-throughput data analysis. This facilitates downstream investigation of the impacts of root hair genetic control and morphological variation on plant performance. pyRootHair is installable via PyPI (https://pypi.org/project/pyRootHair/) and can be accessed on GitHub at https://github.com/iantsang779/pyRootHair.
Why it matches plant phenotyping methods植物根毛形態を顕微鏡画像から自動抽出するAIソフトウェアを開発し、複数作物で適用・実証しており、表現型取得手法が研究の中心である。
abstractWe present pyRootHair, a high-throughput, AI-powered software application to automate root hair trait extraction from microscope images of plant roots grown on agar plates.
Reproduction assets foundThe paper's root hair phenotyping software (pyRootHair) is publicly available on GitHub and PyPI, the data and notebooks used to generate the manuscript figures are deposited in the repository's paper_data folder, and the software is annotated in the DOME-ML registry. The GigaDB deposit (10.5524/102771) is referenced,但Code · publicregression lines were computed using statsmodels (v0.14.4). Scikit-learn (v.1.5.2) was used for quality control of segmented images. nnU-Netv2 (v2.5.1) was used to create the image segmentation model with PyTorch (v.2.5.1) and CUDA (v.12.6).
Availability of Source Code and Requirements
Project name: pyRootHair
Project homepage: https://github.com/iantsang779/pyRootHair
Operating system(s): Linux, MacOS, Windows
Programming language: Python
License: MIT License
Supplementary Material
giaf141_Supplemental_File giaf141_Authors_Response_To_Reviewer_Comments_Original_Submission giaf141_GIGA-D-25-00279_Original_Submission giaf141_GIGA-D-25-00279_Revision_1 giaf141_Reviewer_1_Report_Original_SubmisOpen asset ↗github.com/iantsang779/pyRootHairlines:250-287Dataset · publicThe source jupyter notebook and data used to generate all figures in the manuscript have been deposited on GitHub [ 39 ].Open asset ↗lines:400-405Code · publiclarge number of plant germplasm resources for variation in root hair morphology, supporting high-resolution measurements and high-throughput data analysis. This facilitates downstream investigation of the impacts of root hair genetic control and morphological variation on plant performance. pyRootHair is installable via PyPI ( https://pypi.org/project/pyRootHair/ ) and can be accessed on GitHub at https://github.com/iantsang779/pyRootHair .
Keywords: root hairs, plant phenotyping, machine learning, computer vision, AI, U-Net, wheat, roots, software
status released display-pdf yes is-olf no is-manuscript no is-preprint no is-journal-matter no is-scanned no is-retracted no
Received 2025 JOpen asset ↗lines:1-34Code / dataset availability confirmedEurope PMC · bioRxiv · Crossref · checked 14 Sept 2026
Volume electron microscopy (vEM) provides nanometer-scale, three-dimensional imaging of cells, but applying it to plant systems remains challenging. Cell walls, large vacuoles, and tissue thickness complicate sample preparation and cryogenic imaging. Here we report a cryogenic vEM (cryo-vEM) workflow for unstained plant protoplasts that achieves volumetric imaging of whole vitrified sorghum stem protoplasts without chemical fixation, dehydration, resin embedding, or heavy-metal staining. The method integrates optimized protoplast isolation, plunge-freezing vitrification for native-state preservation, automated cryogenic focus ion beam scanning electron microscopy (cryo-FIB-SEM) slice-and-view acquisition, contrast enhancement and stack alignment, and AI-assisted human-in-the-loop 3D segmentation. Using sorghum stem protoplasts as a demonstration, the workflow captured large-volume frozen-hydrated protoplast ultrastructure, allowing visualization of major organelles, including the nucleus, mitochondria, vacuoles, ER/Golgi-like membranes, lipid bodies, and subcellular features consistent with nuclear-envelope pores. We further quantified organelle volumes and surface areas from the segmented 3D data, highlighting the potential for quantitative cellular ultrastructure analysis. This cryo-vEM workflow provides a platform for near-native structural studies of isolated plant protoplasts.
Why it matches plant phenotyping methods植物プロトプラストの三次元画像取得・セグメンテーション・オルガネラ形態量化を中核とする手法開発であり、植物の細胞形態形質を抽出するため。
abstractHere we report a cryogenic vEM (cryo-vEM) workflow for unstained plant protoplasts that achieves volumetric imaging of whole vitrified sorghum stem protoplasts
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Code · public369 The codes are freely available at https://github.com/xzhang0123/vEMOpen asset ↗https://github.com/xzhang0123/vEM · xzhang0123/vEMpdf-page:10 lines:1-24Code / dataset availability confirmedEurope PMC · checked 6 Sept 2026
Polyploidy (whole-genome duplication) is a common yet under-surveyed property of tissues across multicellular organisms. Polyploidy plays a critical role during tissue development, following acute stress, and during disease progression. Common methods to reveal polyploidy involve either destroying tissue architecture by cell isolation or tedious identification of individual nuclei in intact tissue. Therefore, there is a critical need for rapid and high-throughput ploidy quantification using images of nuclei in intact tissues. Here, we present iSPy (inferring Spatial Ploidy), an unsupervised learning pipeline that is designed to create a spatial map of nuclear ploidy across a tissue of interest. We demonstrate the use of iSPy in Arabidopsis, Drosophila, and human tissue. iSPy can be adapted for a variety of tissue preparations, including whole mount and sectioned. This high-throughput pipeline will facilitate rapid and sensitive identification of nuclear ploidy in diverse biological contexts and organisms.
Why it matches plant phenotyping methodsiSPyは画像から組織内の核倍数性を空間的・高スループットに推定する教師なし学習パイプラインであり、Arabidopsisで実証されている。植物の状態を抽出する計算フェノタイピング手法が中心である。
abstractHere, we present iSPy (inferring Spatial Ploidy), an unsupervised learning pipeline that is designed to create a spatial map of nuclear ploidy across a tissue of interest.
Reproduction assets foundThe paper deposits its paper-specific phenotyping assets publicly: confocal images of A. thaliana, D. melanogaster, and human cardiomyocytes, ilastik segmentation files, and A. thaliana cotyledon flow cytometry data are all in an OSF repository, and the iSPy analysis code is available both on OSF and in a public GitLabDataset · publicAll data presented in the study are publicly available in the OSF data repository (https://osf.io/um7r3/; https://doi.org/10.17605/osf.io/um7r3).Open asset ↗10.17605/osf.io/um7r3html-lines:253-271Code · publicThe code for iSPy can also be found in the OSF data repository (https://osf.io/um7r3/; https://doi.org/10.17605/osf.io/um7r3), as well as in a GitLab repository, https://gitlab.gwdg.de/devplantpatterning/Publications/ispy-inferring-spatial-ploidy.Open asset ↗gitlab.gwdg.de · devplantpatterning/Publications/ispy-inferring-spatial-ploidyhtml-lines:253-271Code / dataset availability confirmedCrossref · Europe PMC · checked 14 Sept 2026
Abstract Mass spectrometry imaging (MSI) is a vital tool in botanical research. Image fusion is introduced for resolution enhancement of MSI data from animal samples, but its application to plant MSI data resulted in unsatisfactory visualizations due to the distinct morphological characteristics of plant tissues. Herein, this study presents loss controlled residual network (LCRN), a workflow dedicated to the super‐resolution fusion of plant MSI data. The pipeline used a residual connection‐based neural network implemented with a novel loss metric called edge perceptual loss. Edge perceptual loss is developed for evaluating complex morphological information that can not be properly reflected by common image metrics, and its implementation in loss propagation is vital to the quality of the fusion result. Compared to existing deep learning‐based methods, LCRN is able to generate a high‐quality super‐resolution fusion image of extra high magnification (up to 20‐fold) that combined chemical and morphological information obtained from MSI and microscopy, respectively.
Why it matches plant phenotyping methods植物組織のMSIデータを対象に、化学情報と形態情報を統合して超解像画像を生成する画像融合ワークフローを開発しており、植物形態の取得・抽出手法が研究の中心である。
abstractHerein, this study presents loss controlled residual network (LCRN), a workflow dedicated to the super‐resolution fusion of plant MSI data.
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Code · publicThe data that support the findings of this study are available in the supplementary material of this article. Codes are available at https://github.com/codexyster/LCRN‐pr .Open asset ↗codexyster/LCRN‐prlines:245-245Code / dataset availability confirmedEurope PMC · checked 6 Sept 2026
Mycorrhizal associations between fungi and plants are a fundamental aspect of terrestrial ecosystems. Mycorrhizas occur in c. 85% of extant plants, yet their geological record remains sparse. Rare fossil evidence from early terrestrial environments offers crucial insights into these ancient symbioses, but visualizing fossil fungi at the microscale within plant tissues is challenging. Here, we combine confocal laser scanning microscopy and fluorescence lifetime imaging microscopy (FLIM) to investigate a newly identified fungus and cellular structures of a 407-Myr-old plant from the Windyfield Chert, a stratigraphically distinct fossiliferous unit from Rhynie (Scotland). We also applied Raman spectroscopy to investigate the carbon framework of both fungal and plant tissues. This integrative approach revealed fungal structures in unprecedented detail. The fungus, Rugososporomyces lavoisierae gen. nov., sp. nov., exhibits features resembling extant Glomeromycotina arbuscular mycorrhizal fungi. This is the first record of mycorrhizas from the Windyfield Chert. FLIM further distinguished features at the subcellular level, while Raman spectroscopy showed that fungal arbuscules and vesicles of the plant water-conducting cells underwent geological alterations, resulting in a similar chemical composition. These findings expand our understanding of ancient and extremely rare plant-fungal symbioses and highlight the potential of confocal-FLIM for advancing palaeobotanical research.
Why it matches plant phenotyping methods植物組織内の微細構造を対象に、共焦点レーザー顕微鏡・FLIM・ラマン分光を組み合わせた観察法を中核としており、化石植物の細胞・菌根構造の状態を抽出している。
abstractHere, we combine confocal laser scanning microscopy and fluorescence lifetime imaging microscopy (FLIM) to investigate a newly identified fungus and cellular structures of a 407-Myr-old plant
Reproduction assets foundThe authors deposited all confocal imaging data used in this fossil mycorrhiza study (CLSM/FLIM datasets of Rugososporomyces lavoisierae in Aglaophyton majus) in a public Zenodo repository under a CC BY 4.0 license. This is a paper-specific, publicly accessible dataset of the phenotyping/imaging measurements.Dataset · publicAll confocal data collected and used in this study are deposited in the Zenodo repository under a Creative Commons Attribution 4.0 international license https://doi.org/10.5281/zenodo.15194427 (Strullu‐Derrien et al ., 2025 ).Open asset ↗Zenodo · 10.5281/zenodo.15194427lines:252-551Code / dataset availability confirmedEurope PMC · Crossref · checked 6 Sept 2026
Canola is an important economic and agronomic crop globally, but its yield is under threat due to climate change. Stomata are a key breeding target because of their importance in carbon capture and water use efficiency. However, screening for elite stomatal traits could be laborious and time-consuming. We developed a new toolkit called Stomatal Comprehensive Automated Neural Network or SCAN that combines the use of high-resolution portable digital microscopy with machine learning to automate stomatal trait phenotyping in canola. We show that SCAN can rapidly measure stomatal density, size, and pore area in canola at 97-99% accuracy, and capture real-time stomatal pore status that strongly correlated with leaf porometer measurement in canola. Here we use SCAN to investigate how leaf stomatal traits vary through a canopy in different ecotypes of canola grown in the field and glasshouse conditions. SCAN revealed that stomatal density in canola decreases in more expanded leaves with the abaxial surface having up to 40% more stomata that are 2× more open than the adaxial surface. SCAN also showed that patterns of stomatal traits in canola vary between leaf position in the canopy and change with environment in an ecotype-dependent manner.
Why it matches plant phenotyping methods葉の気孔形質を自動取得する画像・機械学習ツールを開発し、精度検証と既存測定との相関評価を行っており、植物表現型取得法が研究の中心である。
abstractWe developed a new toolkit called Stomatal Comprehensive Automated Neural Network or SCAN that combines the use of high-resolution portable digital microscopy with machine learning to automate stomatal trait phenotyping in canola.
Reproduction assets foundThe paper publicly releases its authors' analysis code, trained model weights, and training image datasets for the SCAN stomatal phenotyping pipeline via two GitHub repositories and two Roboflow datasets, with explicit availability statements in the Data availability section. Raw phenotype measurements are in a journalCode · publicThe full details of the weights, hyperparameters, training scripts, and datasets of the models can be found at https://github.com/William-Yao0993/FD_detection .Open asset ↗William-Yao0993/FD_detectionlines:45-53Dataset · publicThe microscopy images used to train the SCAN model are available as two public datasets: https://app.roboflow.com/danila-lab/fd-project-1 and https://app.roboflow.com/danila-lab/pore-segmentation/ .Open asset ↗danila-lab/fd-project-1lines:233-272Dataset · publicThe microscopy images used to train the SCAN model are available as two public datasets: https://app.roboflow.com/danila-lab/fd-project-1 and https://app.roboflow.com/danila-lab/pore-segmentation/ .Open asset ↗danila-lab/pore-segmentationlines:233-272Code / dataset availability confirmedEurope PMC · bioRxiv · checked 14 Sept 2026
Background The plant vacuole arises by orchestrated interplay of membrane trafficking, cytoskeletal rearrangements and a variety of signalling pathways. In the root, the characteristic large central vacuole develops by endomembrane reorganization occurring mainly in the transition zone. The vacuole’s bounding membrane - the tonoplast - can be visualized in vivo using fluorescent protein markers, allowing for quantitative analysis of confocal microscopy images. Tonoplast organization can thus serve as a sensitive indicator of changes to any of the processes involved in vacuole biogenesis. The Vacuolar Morphology Index (VMI) is widely accepted as a quantitative measure of vacuole structure. However, this metric has two drawbacks - it only reflects the size of the largest vacuolar compartment (missing therefore possible differences in the organization of smaller compartments), and its determination is labor intensive, limiting its use on large datasets. Results We developed an alternative metric for describing vacuole organization, named the Tonoplast Topology Index (TTI), which overcomes the above-mentioned shortcomings of the VMI. We compared the performance of our protocol with VMI on a simulated dataset and on real data. To validate the methods’ performance, we used it to confirm the previously reported differences in vacuole shape and size between Arabidopsis thaliana roots grown on the surface of an agar medium compared to those embedded inside the agar. Both VMI and TTI could efficiently detect the relatively subtle changes in vacuole organization depending on the position of the root in the agar, and provided correlated results. However, only TTI produced data with close to normal value distribution, simplifying subsequent statistical evaluation. Conclusions We present the protocol for TTI determination as a two-stage semi-automated procedure involving microscopic image analysis employing an ImageJ macro and subsequent processing of numeric data in the Jupyter Notebook environment, together with benchmarking image data. Since this implementation is freeware-based, platform-independent and (relatively) user-friendly, we hope it will find its use as a high throughput, added value alternative to the VMI metric.
Why it matches plant phenotyping methods植物の液胞構造を定量化する新規指標と半自動画像解析プロトコルを開発し、既存指標との比較・実データおよびシミュレーションによる検証、ベンチマークデータを提示しており、表現型取得・抽出法が中心である。
abstractWe developed an alternative metric for describing vacuole organization, named the Tonoplast Topology Index (TTI), which overcomes the above-mentioned shortcomings of the VMI.
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Code · publicthe software tool generated here are also available at https://github.com/GeorgeCaldarescu/TTI-Open asset ↗GeorgeCaldarescu/TTI-pdf-page:9 lines:1-52Code / dataset availability confirmedCrossref · Europe PMC · checked 15 Sept 2026
Abstract Correlative imaging is a powerful tool for revealing information on cell-type structures and their biochemistry, with the potential to inform healthier food choices and improved dietary recommendations. Determination of plant structures and their structural biochemistry advances our understanding of specific structures designed to store different biomolecules within cells and tissues. Compared to the classical biochemical separation techniques, the key advantage of sequential correlative imaging techniques is in relating spatial plant (micro)structures to their biochemistry in a nondestructive manner. Sequential imaging reported here comprises six methodologies on a single sample, a cross-section of a Tartary buckwheat (Fagopyrum tataricum) grain, namely, bright-field and autofluorescence microscopy, fluorescence microspectroscopy, MeV-secondary ion mass spectrometry, micro-particle-induced X-ray emission, scanning electron microscopy coupled with energy dispersive X-ray spectroscopy, and laser ablation-inductively coupled plasma-mass spectrometry. Results confirm that the stepwise addition of the desired information across several classes of biomolecules and several spatial scales informs the quality and safety of plant-based produce across scales. Therefore, a viable workflow is proposed, enabling sequential spatial analysis of grain and highlighting plant structures' in situ specificity. The advantages and disadvantages of the selected methodologies were critically evaluated.
Why it matches plant phenotyping methods植物粒の構造とその化学的特徴を複数の相関イメージング法で取得する再利用可能なワークフローを提案し、各手法の長短も評価しているため、表現型取得法が中心である。
abstractTherefore, a viable workflow is proposed, enabling sequential spatial analysis of grain and highlighting plant structures' in situ specificity.
Reproduction assets foundThe paper explicitly points to a public Zenodo deposit containing the correlative imaging data (SEM, micro-PIXE, MeV-SIMS maps) used in its analyses, with instructions for reproducing image fusion in Wolfram Mathematica or ImageJ.Dataset · publicsed to reveal the allocation of K
to cotyledons (Supplementary Fused Image 1). Similarly, on
the same SEM image, MeV-SIMS distribution maps under the
selected peak were overlaid (Supplementary Fused Image 2).
Custom combinations can be done in the Wolfram
Mathematica program or in ImageJ (Merge Channels) using
data available at https://doi.org/10.5281/zenodo.14628251, fol
lowing the instructions in the Materials and Methods.
Conclusions
The low emission properties of fluorescence biomolecules,
when excited with 405 nm light, inherently limit the informa
tion acquired using fluorescence imaging. At this excitation
wavelength, catechin may be the primary fluorophore in
Tartary buckwheat cotOpen asset ↗zenodo · 10.5281/zenodo.14628251pdf-raw-page:13 lines:1-89Code / dataset availability confirmedOpenAlex · Europe PMC · checked 6 Sept 2026
L.), a vital global food crop, relies on its stomatal structure for regulating photosynthesis and responding to drought. Conventional manual stomatal detection methods are inefficient, subjective, and inadequate for high-throughput plant phenotyping research. To address this, we curated a dataset of over 1500 maize leaf epidermal stomata images and developed a novel lightweight detection model, StomaYOLO, tailored for small stomatal targets and subtle features in microscopic images. Leveraging the YOLOv11 framework, StomaYOLO integrates the Small Object Detection layer P2, the dynamic convolution module, and exploits large-scale epidermal cell features to enhance stomatal recognition through auxiliary training. Our model achieved a remarkable 91.8% mean average precision (mAP) and 98.5% precision, surpassing numerous mainstream detection models while maintaining computational efficiency. Ablation and comparative analyses demonstrated that the Small Object Detection layer, dynamic convolutional module, multi-task training, and knowledge distillation strategies substantially enhanced detection performance. Integrating all four strategies yielded a nearly 9% mAP improvement over the baseline model, with computational complexity under 8.4 GFLOPS. Our findings underscore the superior detection capabilities of StomaYOLO compared to existing methods, offering a cost-effective solution that is suitable for practical implementation. This study presents a valuable tool for maize stomatal phenotyping, supporting crop breeding and smart agriculture advancements.
Why it matches plant phenotyping methodsトウモロコシの気孔を画像から検出するモデルとデータセットを開発・評価しており、植物フェノタイピング手法が研究の中心です。
abstractwe curated a dataset of over 1500 maize leaf epidermal stomata images and developed a novel lightweight detection model, StomaYOLO
Reproduction assets foundThe paper's analysis code (StomaYOLO detector) is openly available on GitHub with an authors' URL; the phenotype image dataset itself is only available on request from the corresponding author.Code · publicThe code that support the findings of this study are openly available in GitHub at https://github.com/yangziqi2003/StomaYOLO (accessed on 5 May 2025).Open asset ↗https://github.com/yangziqi2003/StomaYOLO · StomaYOLOlines:426-440Code / dataset availability confirmedEurope PMC · bioRxiv · checked 6 Sept 2026
Summary Live imaging data analysis often requires an objective, local, and accurate way of quantification of cell dynamics. In the research field of polarized tip-growth, the cell fluctuations and/or fluctuations in tip position and growth direction hampers automated analyses of huge amounts of imaging sequences. The fluctuated nature in data makes it unclear how cell shape and growth are linked to intracellular events that could be the actual driving force of cell growth. To overcome these difficulties, we developed a powerful and user-friendly tool called KymoTip with an available format. In this software, novel functions such as coordinate normalization, tip-bottom detection, and signal kymograph were implemented. We confirmed that not only plasma membrane-labeled fluorescent images, but also images such as bright-field and cortical microtubule markers —so long as the cell contours can be identified— are amenable to KymoTip. Furthermore, by combining markers for cell contours with those that visualize intracellular structures, it becomes possible to quantitatively analyze various intracellular events, such as nuclear migration and calcium wave, in conjunction with cellular growth dynamics. Since KymoTip can be handled by non-specialist, it is expected to promote understanding of what happens at the sub- and cellular level with high throughput outcomes. Significance statement Faced with fluctuations in cell coordinates and cell tip positions, position correction of live imaging data and accurate detection of tip position are key challenges in plant developmental biology. We solved them with a powerful and user-friendly tool, KymoTip, that can realize cell position correction, cell tip detection with cell centerline, and quantification of intracellular events.
Why it matches plant phenotyping methods植物細胞のライブイメージから細胞形状・先端位置・成長動態を定量化する解析ソフトウェアを開発しており、植物フェノタイピング手法が中心です。
abstractwe developed a powerful and user-friendly tool called KymoTip
Reproduction assets foundThe paper's authors explicitly state that the KymoTip analysis code is publicly available on GitHub at https://github.com/blues0910/KymoTip, which is an allowed URL. This is the authors' own computational tool implementing the paper's tip-growth phenotyping analysis (segmentation, coordinate normalization, tip-bottom, Code · publicThe code for KymoTip is available on GitHub: https://github.com/blues0910/KymoTip.Open asset ↗blues0910/KymoTippdf-page:8 lines:1-44Code / dataset availability confirmedbioRxiv · checked 15 Sept 2026
The global carbon cycle depends heavily on the carbon sequestration rates of aquatic ecosystems. Sinking of phytoplankton is a rapid mediator of carbon sequestration, because phytoplankton are globally abundant photoautotrophs that grow rapidly. Pico- and nano-phytoplankton sinking velocities vary depending on their growth state, viability, clumping, and distribution in the water column. We introduced high throughput fluorescence microscopy of well-plates, to measure sinking velocities of three diatom strains, and three cyanobacteria strains, with cell radii spanning an order of magnitude, all grown under three different light levels. Cultures were measured for sinking velocities repeatedly across their growth trajectories. Tracking multiple fluorescence wavebands allowed us to simultaneously determine sinking velocities for living vs. dead cells. Sinking velocities varied strongly across growth light levels, and across growth stages. These monoclonal cultures furthermore show distinct sub-populations of slow- and fast-sinking cells. Our results departed widely from simple Stokes Law estimates of sinking based upon radii and mass density of cells. Complex, heterogeneous phytoplankton communities likely show more complicated sinking patterns than are currently expressed in biogeochemical ocean models. Our well-plate microscopy approach using parallel imaging of many samples generates high-throughput measures of cell sinking at population- or community-scales, to in turn improve modelling of carbon export to deeper layers.
Why it matches plant phenotyping methods植物プランクトンの沈降速度を高スループット蛍光顕微鏡で測定する手法を導入し、生活状態や集団スケールの生理・機能形質を定量化しているため、測定法が研究の中心です。
abstractWe introduced high throughput fluorescence microscopy of well-plates, to measure sinking velocities of three diatom strains, and three cyanobacteria strains
Reproduction assets foundThe paper's sinking-velocity analysis code is explicitly stated to be openly available on the authors' GitHub repository. The raw phenotype data is promised for Dryad only upon acceptance, so it is not yet publicly actionable.Code · publicfunctional groups of cyanobacteria, diatoms strains with 156
diameter less than 10µm and diatoms strains with diameter larger than 10µm based on 157
growth light, viability state (living vs. dead and dying) and slow vs. fast sinking 158
velocity clustering groups. 159
The code used to analyse the data is public available at 160
https://github.com/maxberthold/PhytoplanktonSinkVelocities. 161
Sinking according to Stokes’ law 162
Sinking velocities of spherical objects falling under the case of Reynolds numbers 163
smaller than 1 can be described by Stokes’ law. Several studies have used Stokes law or 164
a modified version of Stokes’ law to estimate sinking velocities of plankton and marine 16Open asset ↗maxberthold/PhytoplanktonSinkVelocitiespdf-raw-page:8 lines:1-44Code / dataset availability confirmedEurope PMC · Crossref · checked 14 Sept 2026
Chloroplast density in cells varies among different types of cells and plants. In current single-cell spatiotemporal analysis, the automatic detection and quantification of chloroplasts at the single-cell level is crucial. We developed DeepD&Cchl (Deep-learning-based Detecting-and-Counting-chloroplasts), an AI tool for single-cell chloroplast detection and cell-type clustering. It utilizes You-Only-Look-Once (YOLO), a real-time detection algorithm, for accurate and efficient performance. DeepD&Cchl has been proved to identify chloroplasts in plant cells across various imaging types, including light microscopy, electron microscopy, and fluorescence microscopy. Integrated with an Intersection Over Union (IOU) module, DeepD&Cchl precisely counts chloroplasts in single- or multi-layered images, while eliminating double-counting errors. Furthermore, when combined with Cellpose, a single-cell segmentation tool, DeepD&Cchl enhances its effectiveness at the single-cell level. By counting chloroplasts within individual cells, it supports cell-type-specific clustering based on chloroplast number versus cell size, offering valuable morphological insights for single-cell studies. In summary, DeepD&Cchl is a significant advancement in plant cell analysis. It offers accuracy and efficiency in chloroplast identification, counting and cell-type classification, providing a useful tool for plant research.
Why it matches plant phenotyping methods植物細胞画像から葉緑体を検出・計数し、細胞型をクラスタリングするAIツールの開発が中心であり、植物の形態的状態を定量化するフェノタイピング手法に該当する。
abstractWe developed DeepD&Cchl (Deep-learning-based Detecting-and-Counting-chloroplasts), an AI tool for single-cell chloroplast detection and cell-type clustering.
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Code · publicThe raw dataset, as well as the scripts for the DeepD&Cchl model training and 17application macro, were shared on GitHub https://github.com/Shaokai9/AI4LifeScience_ECNU/tree/main/Deep%20subcellular%20detection .Open asset ↗Shaokai9/AI4LifeScience_ECNUlines:400-410Code / dataset availability confirmedEurope PMC · checked 6 Sept 2026
Grana are fundamental structural units of the intricate chloroplast membrane network. Investigating their nanomorphology is essential for understanding photosynthetic efficiency regulation. Here, we present GRANA (Graphical Recognition and Analysis of Nanostructural Assemblies), an artificial intelligence-enhanced, user-friendly software tool that recognizes grana on thylakoid network electron micrographs and generates a complex set of their structural parameters. GRANA employs 3 artificial neural networks of different architectures and binds them in a 1-click workflow. Its output is designed to facilitate hybrid intelligence analysis, securing fast and reliable results from large datasets. The GRANA tool is over 100 times faster compared with currently used manual approaches. As a proof of concept, we have successfully applied GRANA software to diverse grana structures across different land plant species grown under various conditions, demonstrating the wide range of potential applications for our software. GRANA tool supports large-scale analysis of grana nanomorphological features, facilitating advancements in photosynthesis-oriented studies.
Why it matches plant phenotyping methods葉緑体グラナの電子顕微鏡画像から構造パラメータを自動抽出するソフトウェアの開発であり、植物形態形質の取得・解析法が中心。
abstractan artificial intelligence-enhanced, user-friendly software tool that recognizes grana on thylakoid network electron micrographs and generates a complex set of their structural parameters.
Reproduction assets foundThe paper's raw TEM images used for grana nanomorphology analysis are publicly deposited under DOI 10.58132/HTWCC1. The authors' analysis code (github.com/center4ml/GRANA) is mentioned but that URL is not among the allowed URLs, so it cannot be listed.Dataset · publicRaw TEM data used for results in the manuscript are available at https://doi.org/10.58132/HTWCC1 .Open asset ↗10.58132/HTWCC1lines:184-235Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
Premise The quantification of plant drought resistance, particularly embolism formation, within and across species, is critical for ecosystem management and agriculture. We developed a cost-effective protocol to measure the water potential at which 50% of hydraulic conductivity ( P 50 ) is lost in stems, using affordable and accessible materials in comparison to the traditional optical method. Methods and results Our protocol uses inexpensive USB microscopes, which are secured along with the plants to a pegboard base to avoid movement. A Python program automatized the image acquisition. This method was applied to quantify P 50 in an exotic species ( Nicotiana glauca ) and native species ( Rhus integrifolia ) of the Mediterranean vegetation in Baja California, Mexico. Conclusions The intra- and interspecific patterns of variation in stem P 50 of N. glauca and R. integrifolia were obtained using the low-cost optical method with widely available and affordable materials that can be easily replicated for other species.
Why it matches plant phenotyping methods植物の茎の水理的脆弱性（P50）を測定する低コスト光学プロトコルを開発し、USB顕微鏡とPythonによる画像取得を用いて適用・検証しており、表現型取得法が研究の中心である。
abstractWe developed a cost-effective protocol to measure the water potential at which 50% of hydraulic conductivity ( P 50 ) is lost in stems, using affordable and accessible materials in comparison to the traditional optical method.
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Code · publicGranados (CICESE) for the initial design of the microscope stands, and Alexis Crespo Michel (CICESE) for his assistance in developing the multi‐threaded version of the image capture Python program.
DATA AVAILABILITY STATEMENT
Data of all experiments are provided in the Supporting Information. The Python Program is available at: https://github.com/miguel-aalonso/lowcost_P50 .
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163 : 451 – 452 .
33873751
10.1111/j.1469-8137.2004.01142.x
Avila , R. T.
,
A. A.
Cardoso
,
T. A.
Batz
,
C. N.
Kane
,
F. M.
DaMatta
, and
S. A.
McAdaOpen asset ↗miguel-aalonso/lowcost_P50lines:264-337Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
The thylakoid membrane is the site of the light-dependent reactions of photosynthesis. It is a continuous membrane, folded into grana stacks and the interconnecting stroma lamellae. The CURVATURE THYLAKOID1 (CURT1) protein family is involved in the folding of the membrane into the grana stacks. The thylakoid membrane remodels its architecture in response to light conditions, but its 3D organisation and dynamics remain incompletely understood. To resolve these details, an imaging technique is needed that provides high-resolution 3D images in a high-throughput manner. Recently, we have used expansion microscopy, a technique that meets these criteria, to visualise the thylakoid membrane isolated from spinach. Here, we show that this protocol can also be used to visualise enveloped spinach chloroplasts. Additionally, we present an improved protocol for resolving the thylakoid structure of Arabidopsis thaliana. Using this protocol, we show the changes in thylakoid architecture in response to long-term far-red light acclimation and due to knocking out CURT1A. We show that far-red light acclimation results in higher grana stacks that are packed closer together. In addition, the distance between stroma lamellae, which are wrapped around the grana, decreases. In the curt1a mutant, grana have an increased diameter and height, and the distance between grana is increased. Interestingly, in this mutant, the stroma lamellae occasionally approach the grana stacks from the top. These observations show the potential of expansion microscopy to study the thylakoid membrane architecture.
Why it matches plant phenotyping methods植物のチラコイド膜構造を高解像度3D画像で取得する拡大顕微鏡法の改良・適用が中心であり、膜構造という植物形態形質を測定しているため。
abstractTo resolve these details, an imaging technique is needed that provides high-resolution 3D images in a high-throughput manner.
Reproduction assets foundThe article states that the data underlying the publication (expansion microscopy imaging/measurements of thylakoid architecture) are publicly available in the 4TU Research Data repository via the DOI 10.4121/75fa3c66-8505-4d6a-9bd9-16973e5ca885. This is a paper-specific, publicly accessible data deposit with an authorDataset · publicUte Armbruster for providing the
seeds of the Ler0 curt1a-1 mutant. This work was supported by the Dutch
Organisation for Scientific Research (NWO) via a Vidi grant no. VI.Vidi
192.042 (E.W.) and by Wageningen Graduates Schools through a PhD
grant (J.B.).
Data availability
The data underlying this publication can be accessed at https://doi.org/10.4121/75fa3c66-8505-4d6a-9bd9-16973e5ca885.References
[1] R.E. Blankenship, Molecular Mechanisms of Photosynthesis, John Wiley & Sons,
2021, https://doi.org/10.1002/9780470758472.
[2] H. Kirchhoff, Chloroplast ultrastructure in plants, New Phytol. 223 (2) (2019)
565–574, https://doi.org/10.1111/nph.15730.
[3] H. Kirchhoff, C. Hall, M. Wood, M. HerbstOpen asset ↗10.4121/75fa3c66-8505-4d6a-9bd9-16973e5ca885pdf-raw-page:9 lines:1-68Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
Published1 Mar 2025The Plant journal : for cell and molecular biologyCited by 6 · OpenAlex ↗
High lipid producing (HLP) tobacco (Nicotiana tabacum) is a potential biofuel crop that produces an excess of 30% dry weight as lipid bodies in the form of triacylglycerol. While using HLP tobacco as a sustainable fuel source is promising, it has not yet been tested for its tolerance to warmer environments that are expected in the near future as a result of climate change. We found that HLP tobacco had reduced stomatal conductance, which results in increased leaf temperatures up to 1.5°C higher under control and high temperature (38°C day/28°C night) conditions, reduced transpiration, and reduced CO 2 assimilation. We hypothesize this reduction in stomatal conductance is due to the presence of excessive, large lipid droplets in HLP guard cells imaged using confocal microscopy. High temperatures also significantly reduced total fatty acid levels by 55% in HLP plants; thus, additional engineering may be needed to maintain high titers of leaf oil under future climate conditions. High-throughput image analysis techniques using open-source image analysis platform PlantCV for thermal image analysis (plant temperature), stomata microscopy image analysis (stomatal conductance), and fluorescence image analysis (photosynthetic efficiency) were developed and applied in this study. A corresponding set of PlantCV tutorials are provided to enable similar studies focused on phenotyping future crops under adverse conditions.
Why it matches plant phenotyping methodsPlantCVを用いた熱画像・気孔顕微鏡画像・蛍光画像の高スループット解析手法を開発・適用し、植物温度、気孔関連指標、光合成効率を推定しているため、表現型取得手法が中心的です。
abstractHigh-throughput image analysis techniques using open-source image analysis platform PlantCV for thermal image analysis (plant temperature), stomata microscopy image analysis (stomatal conductance), and fluorescence image analysis (photosynthetic efficiency) were developed and applied in this study.
Reproduction assets foundThe paper's raw phenotyping image data (thermal, fluorescence, stomata, confocal microscopy) are deposited on Zenodo, and the authors' PlantCV analysis workflows and R scripts are on GitHub, including three PlantCV tutorials for thermal, stomata, and photosynthesis analysis.Dataset · publicaxial side of the leaf rather than a cross section. While small lipid droplets were present in the WT stomatal guard cells and epidermis, large lipid droplets were present in the HLP guard cells under both control and after 7 days of treatment (representative control images in Figure 8A–D , complete dataset available on Zenodo, https://zenodo.org/records/10711864 ). In addition, while HLP oil appeared to form spherical droplets, it did not “line” the stomatal opening as in WT (Figure 8C,D ).
Figure 8
High lipid producing (HLP) had excessive oil droplets in stomatal guard cells.
Representative confocal microscopy images, shown as focused Z‐stack, of tobacco leaf tissue fixed in paraformaOpen asset ↗Zenodolines:115-123Code · publicmated marginal means (LSMEANS) to determine which sample types were significantly different from others. Means are reported in text with standard error. Plots were made using ggplot2 package (v.3.5.0) in R. Jupyter notebooks associated with PlantCV analyses and R scripts associated with this manuscript are available on Github ( https://github.com/danforthcenter/tobacco‐heat‐paper ).
AUTHOR CONTRIBUTIONS
DKA, MAG, PDB, BSJ and KMM designed experiments. KMM and BSJ performed experiments and data analysis. KJC designed and aided KMM in confocal and brightfield microscopy experiments and advised TEM experiments. JW performed TEM experiments, and KG‐O and SK performed data analysis of TEM images.Open asset ↗GitHublines:171-182Code · publictification was used to isolate only individual plants in each mask. Then, the mask was applied to the registered thermal image to calculate the average plant temperature, as well as a histogram of pixel temperatures for each plant. A PlantCV workflow was used to analyze the images in parallel. A tutorial is available on GitHub: https://github.com/danforthcenter/plantcv‐tutorial‐thermal?tab=readme‐ov‐file (Acosta‐Gamboa et al., 2024 ). Scripts for this project are available at https://github.com/danforthcenter/tobacco‐heat‐paper . Raw image data are available on Zenodo, https://zenodo.org/records/10711864 .
Stomatal aperture measurements
To measure stomatal number and aperture, leaf impressioOpen asset ↗GitHublines:142-146Code · publicpackage was then used to calculate the number of stomata and the area of the aperture. A limitation of this method is that it does not provide the width and length of stomata, or measurements of the guard cells themselves; instead, it provides the aperture area (a result of length and width). A tutorial is available on GitHub: https://github.com/danforthcenter/plantcv‐stomata‐tutorial‐pcv4 (Murphy, 2024 ). Scripts for this project are available at https://github.com/danforthcenter/tobacco‐heat‐paper . Raw image data are available on Zenodo, https://zenodo.org/records/10711864 .
Photosynthesis and gas exchangeOpen asset ↗GitHublines:142-146Code · publicPlantCV (Gehan et al., 2017 ) using the photosynthesis package; the chlorophyll fluorescence image was used to mask the image for only plant pixels, and average F
v / F
m , F q ′ / F m ′ , NPQ, chlorophyll index, and anthocyanin index were calculated as an average per plant at each timepoint. A tutorial is available on GitHub: https://github.com/danforthcenter/plantcv‐tutorial‐photosynthesis?tab=readme‐ov‐file (Schuhl et al., 2024 ). Scripts for this project are available at https://github.com/danforthcenter/tobacco‐heat‐paper . Raw image data are available on Zenodo, https://zenodo.org/records/10711864 .
Microscopy imaging of lipids
Leaf samples analyzed for lipid content were taken from thOpen asset ↗GitHublines:156-164Code / dataset availability confirmedCrossref · checked 6 Sept 2026
ArabidopsisMicroscopyCell / cellular structureRootTissueVisualization / data management
Abstract Super-resolution methods provide far better spatial resolution than the optical diffraction limit of about half the wavelength of light (∼200–300 nm). Nevertheless, they have yet to attain widespread use in plants, largely due to plants' challenging optical properties. Expansion microscopy (ExM) improves effective resolution by isotropically increasing the physical distances between sample structures while preserving relative spatial arrangements and clearing the sample. However, its application to plants has been hindered by the rigid, mechanically cohesive structure of plant tissues. Here, we report on whole-mount ExM of thale cress (Arabidopsis thaliana) root tissues (PlantEx), achieving a 4-fold resolution increase over conventional microscopy. Our results highlight the microtubule cytoskeleton organization and interaction between molecularly defined cellular constituents. Combining PlantEx with stimulated emission depletion microscopy, we increase nanoscale resolution and visualize the complex organization of subcellular organelles from intact tissues by example of the densely packed COPI-coated vesicles associated with the Golgi apparatus and put these into a cellular structural context. Our results show that ExM can be applied to increase effective imaging resolution in Arabidopsis root specimens.
Why it matches plant phenotyping methods植物組織に適用可能な超解像イメージング手法を開発し、Arabidopsis根で解像度向上を実証しており、画像取得法が研究の中心である。
abstractHere, we report on whole-mount ExM of thale cress (Arabidopsis thaliana) root tissues (PlantEx), achieving a 4-fold resolution increase over conventional microscopy.
Reproduction assets foundThe paper's PlantEx expansion microscopy imaging data are deposited in ISTA's public repository, and the authors' custom analysis code (including the BigWarp-based expansion-factor script) is publicly available on GitHub. The Click-ExM repository is cited prior work whose method was adapted, not a paper-specific asset.Dataset · publicThe data that support the findings of this study are available via ISTA's data repository at https://doi.org/10.15479/AT:ISTA:18837 .Open asset ↗ISTA's data repository · 10.15479/AT:ISTA:18837lines:219-252Code · publicThe custom-written code used and described in this manuscript is available via Github ( https://github.com/danzllab/PlantEx ).Open asset ↗github.com/danzllab/PlantExlines:219-252Code · publicThe expansion factor was extracted as the linear scaling factor of the similarity transformation minimizing squared landmark residuals using the script https://github.com/danzllab/CATS/tree/master/rcats_image-analysis/bigwarp .Open asset ↗github.com/danzllab/CATSlines:154-159Code / dataset availability confirmedEurope PMC · checked 6 Sept 2026
Hormonal mechanisms associated with cell elongation play a vital role in the development and growth of plants. Here, we report Nextflow-root (nf-root), a novel best-practice pipeline for deep-learning-based analysis of fluorescence microscopy images of plant root tissue from A. thaliana. This bioinformatics pipeline performs automatic identification of developmental zones in root tissue images. This also includes apoplastic pH measurements, which is useful for modeling hormone signaling and cell physiological responses. We show that this nf-core standard-based pipeline successfully automates tissue zone segmentation and is both high-throughput and highly reproducible. In short, a deep-learning module deploys deterministically trained convolutional neural network models and augments the segmentation predictions with measures of prediction uncertainty and model interpretability, while aiming to facilitate result interpretation and verification by experienced plant biologists. We observed a high statistical similarity between the manually generated results and the output of the nf-root.
Why it matches plant phenotyping methods植物根組織の発達ゾーンを画像から自動抽出し、アポプラストpHを測定する再現可能な深層学習パイプラインを開発・検証しており、植物表現型取得が中心である。
abstractThis bioinformatics pipeline performs automatic identification of developmental zones in root tissue images.
Reproduction assets foundThe paper publicly releases the PHDFM fluorescence microscopy image dataset, a test dataset, the trained U-Net^2 segmentation model, the nf-root Nextflow pipeline, the segmentation training module, and the prediction package implementing uncertainty/interpretability, all with explicit availability statements and ZenodoDataset · publicThe PHDFM dataset is available at https://zenodo.org/record/5841376/ .Open asset ↗zenodo · 5841376lines:127-159Dataset · publicthe test dataset for the pipeline ( https://zenodo.org/record/5949352/ ) are publicly available online.Open asset ↗zenodo · 5949352lines:127-159Code · publicsoftware and hardware information are also available in the module ( https://github.com/qbic-pipelines/root-tissue-segmentation-core ). We used version 1.0.1 of the segmentation training module.Open asset ↗github · qbic-pipelines/root-tissue-segmentation-corelines:106-126Code / dataset availability confirmedbioRxiv · checked 13 Sept 2026
Timelapse microscopy has recently been employed to study the metabolism and physiology of cyanobacteria at the single-cell level. However, the identification of individual cells in brightfield images remains a significant challenge. Traditional intensity-based segmentation algorithms perform poorly when identifying individual cells in dense colonies due to a lack of contrast between neighboring cells. Here, we describe a newly developed software package called Cypose which uses machine learning (ML) models to solve two specific tasks: segmentation of individual cyanobacterial cells, and classification of cellular phenotypes. The segmentation models are based on the Cellpose framework, while classification is performed using a convolutional neural network named Cyclass. To our knowledge, these are the first developed ML-based models for cyanobacteria segmentation and classification. When compared to other methods, our segmentation models showed improved performance and were able to segment cells with varied morphological phenotypes, as well as differentiate between live and lysed cells. We also found that our models were robust to imaging artifacts, such as dust and cell debris. Additionally, the classification model was able to identify different cellular phenotypes using only images as input. Together, these models improve cell segmentation accuracy and enable high-throughput analysis of dense cyanobacterial colonies and filamentous cyanobacteria.
Why it matches plant phenotyping methodsシアノバクテリア細胞の画像セグメンテーションと細胞表現型分類を行うソフトウェアおよび機械学習手法の開発が中心であり、植物細胞の形態・生存状態を抽出するフェノタイピング手法に該当する。
abstractHere, we describe a newly developed software package called Cypose which uses machine learning (ML) models to solve two specific tasks: segmentation of individual cyanobacterial cells, and classification of cellular phenotypes.
Reproduction assets foundThe paper's segmentation/classification models and analysis code are publicly available in the authors' GitHub repository (cameronlab/cypose). The microscopy training datasets are not public and are available only upon request.Code · publicAll code and trained models can be downloaded from https://github.com/cameronlab/cypose .Open asset ↗cameronlab/cyposelines:298-383Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
The present dataset combines transcriptomic and microscopic analyses to investigate the responses of winter oilseed rape (WOSR, Brassica napus L., cultivar Aviso) to soil drought, with a focus on differences between young and early-senescent old leaves. For microscopy, 36 scans of 1 to 5 leaf cross-sections were acquired from paraffin-embedded leaf disc samples using a scanner with a 40x lens (Pannoramic Confocal, 3DHistech), capturing a large field of view (8-mm-long observed leaf tissue). The raw scanned cross-sections and analyzed images are available under doi.org/10.57745/RK5PM3 in the Recherche Data Gouvrepository. These high-quality scans enable the differentiation of mesophyll cells and tissues. Software analysis yielded a dataset with 54 selected cross-sectional areas, 291 delimited surfaces of palisade, spongy, and vessel tissues, and 11,136 individually delimited cells from the palisade and spongy layers. For transcriptomics, an Illumina Novaseq sequencer was used to generate 390 Gb of mRNA paired-end reads. The raw reads were filtered, mapped, and assigned to genes from the Brassica napus reference genome Darmor-bzh v10, which were subsequently used to identify differentially expressed genes (DEGs) and to perform gene ontology enrichment analysis. The raw reads are accessible under accession PRJNA939927 at the NCBI Sequence Read Archive (SRA). This high-quality dataset provides insights into the molecular mechanisms underlying oilseed rape's response to soil drought and may aid in the development of drought-tolerant cultivars. A total of 17,975 DEGs were identified between well-watered and severe drought conditions across the contrasted leaf developmental stages.
Why it matches plant phenotyping methods葉の断面画像を取得・解析し、組織面積や個別細胞などの植物形態形質を構造化した再利用可能なデータセットを提供しており、画像ベースの表現型取得が実質的な構成要素である。
abstractFor microscopy, 36 scans of 1 to 5 leaf cross-sections were acquired from paraffin-embedded leaf disc samples using a scanner with a 40x lens
Reproduction assets foundThe article deposits its own plant-phenotyping assets publicly: raw and analyzed leaf cross-section microscopy scans (Recherche Data Gouv, doi:10.57745/RK5PM3) and the transcriptomic dataset (Recherche Data Gouv doi:10.57745/7HQSM3, mirrored at NCBI SRA under PRJNA939927). The analysis pipelines cited (nf-core/rnaseq, Dataset · publicThe raw scanned cross-sections and analyzed images are available under doi.org/10.57745/RK5PM3 in the Recherche Data Gouvrepository.Open asset ↗Recherche Data Gouv · 10.57745/RK5PM3lines:1-41Code / dataset availability confirmedEurope PMC · Crossref · bioRxiv · checked 15 Sept 2026
The applicability of a deep learning model for the virtual staining of plant cell structures using bright-field microscopy was investigated. The training dataset consisted of microscopy images of tobacco BY-2 cells with the plasma membrane stained with the fluorescent dye PlasMem Bright Green and the cell nucleus labeled with Histone-red fluorescent protein. The trained models successfully detected the expansion of cell nuclei upon aphidicolin treatment and a decrease in the cell aspect ratio upon propyzamide treatment, demonstrating its utility in cell morphometry. The model also accurately documented the shape of Arabidopsis pavement cells in both wild type and the bpp125 triple mutant, which has an altered pavement cell phenotype. Metrics such as cell area, circularity, and solidity obtained from virtual staining analyses were highly correlated with those obtained by manual measurements of cell features from microscopy images. Furthermore, the versatility of virtual staining was highlighted by its application to track chloroplast movement in Egeria densa . The method was also effective for classifying live and dead BY-2 cells using texture-based machine learning, suggesting that virtual staining can be applied beyond typical segmentation tasks. Although this method still has some limitations, its non-invasive nature and efficiency make it highly suitable for label-free, dynamic, and high-throughput analyses in quantitative plant cell biology.
Why it matches plant phenotyping methods植物細胞構造の仮想染色と画像解析モデルを開発・評価し、細胞面積・形状・核拡大・葉緑体運動・生死などの表現型を定量化しているため、フェノタイピング手法が中心である。
abstractThe applicability of a deep learning model for the virtual staining of plant cell structures using bright-field microscopy was investigated.
Reproduction assets foundThe paper publicly releases its virtual-staining training/test image sets (bright-field inputs with paired confocal reference images) for BY-2 vacuole, BY-2 nucleus/plasma membrane, and E. densa chloroplast models on figshare under CC BY 4.0, via three DOIs listed in the Data Availability section. No author analysis orDataset · publicata pertaining to this article will be shared on reasonable request to the corresponding author. The
training and test image sets for BY-2 cells and E. densa, which are publicly accessible on figshare under the
CC BY 4.0 license, include images of wild-type tobacco BY-2 cells stained with BCECF for vacuolar lumen
visualization (https://doi.org/10.6084/m9.figshare.27247629.v1), transgenic tobacco BY-2 cells with
.
CC-BY-NC-ND 4.0 International license
perpetuity. It is made available under a
preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in
The copyright holder for this
this version posted October 25, 2024.
;
hOpen asset ↗figshare · 10.6084/m9.figshare.27247629.v1pdf-raw-page:21 lines:1-32Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
Understanding lignocellulosic biomass resistance to enzymatic deconstruction is crucial for its sustainable conversion into bioproducts. Despite scientific advances, quantitative morphological analysis of plant deconstruction at cell and tissue scales remains under-explored. In this study, an original pipeline is devised, involving four-dimensional (space + time) fluorescence confocal imaging, and a novel computational tool, to track and quantify deconstruction at cell and tissue scales. By applying this pipeline to poplar wood, dynamics of cellular parameters was computed and cellulose conversion during enzymatic deconstruction was measured. Results showed that enzymatic deconstruction predominantly impacts cell wall volume rather than surface area. Additionally, a negative correlation was observed between pre-hydrolysis compactness measures and volumetric cell wall deconstruction rate, whose strength was modulated by enzymatic activity. Results also revealed a strong positive correlation between average volumetric cell wall deconstruction rate and cellulose conversion rate. These findings link key deconstruction parameters across nano and micro scales.
Why it matches plant phenotyping methods植物細胞・組織の分解状態を定量する4次元蛍光共焦点イメージングと計算ツールが研究の中心であり、植物状態の形態的変化を抽出する方法を開発している。
abstractIn this study, an original pipeline is devised, involving four-dimensional (space + time) fluorescence confocal imaging, and a novel computational tool, to track and quantify deconstruction at cell and tissue scales.
Reproduction assets foundThe paper's WallTrack computational pipeline (used to track and quantify 4D confocal imaging of poplar cell wall deconstruction) is publicly available on the authors' FARE laboratory GitLab repository. The underlying imaging/phenotype data are not publicly deposited; the authors state data will be made available on.Code · publicnano and micro
scales.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data availability
The WallTrack code is accessible through the FARE laboratory
GitLab repository at: https://gitlab.com/farelab/teamyr/publications/refahi_et_al_4d. Data will be made available on request.
Acknowledgments
The authors thank Anouck Habrant for her help in confocal imaging
and Grégoire Malandain, Solmaz Hossein Khani, Khadidja Ould Amer,
and Ali Faraj for their comments on the manuscript. This work was
supported by Agence Nationale de la Recherche (ANR) Open asset ↗https://gitlab.com/farelab/teamyr/publications/refahi_et_al_4d · refahi_et_al_4dpdf-raw-page:11 lines:1-66Code / dataset availability confirmedEurope PMC · checked 15 Sept 2026
Arabidopsis thaliana sepals are excellent models for analyzing growth of entire organs due to their relatively small size, which can be captured at a cellular resolution under a confocal microscope. To investigate how differential growth of connected cell layers generate unique organ morphologies, it is necessary to live-image deep into the tissue. However, imaging deep cell layers of the sepal (or plant tissues in general) is practically challenging. Image processing is also difficult due to the low signal-to-noise ratio of the deeper tissue layers, an issue mainly associated with live imaging datasets. Addressing some of these challenges, we provide an optimized methodology for live imaging sepals, and subsequent image processing. For live imaging early-stage sepals, we found that the use of a bright fluorescent membrane marker, coupled with increased laser intensity and an enhanced Z- resolution produces high-quality images suitable for downstream image processing. Our optimized parameters allowed us to image the bottommost cell layer of the sepal (inner epidermal layer) without compromising viability. We used a 'voxel removal' technique to visualize the inner epidermal layer in MorphoGraphX image processing software. We also describe the MorphoGraphX parameters for creating a 2.5D mesh surface for the inner epidermis. Our parameters allow for the segmentation and parent tracking of individual cells through multiple time points, despite the weak signal of the inner epidermal cells. While we have used sepals to illustrate our approach, the methodology will be useful for researchers intending to live-image and track growth of deeper cell layers in 2.5D for any plant tissue.
Why it matches plant phenotyping methods植物組織の深部をライブイメージングし、画像処理・細胞セグメンテーション・追跡によって成長を解析する方法自体が中心的に開発・最適化されているため。
abstractwe provide an optimized methodology for live imaging sepals, and subsequent image processing.
Reproduction assets foundThe paper's Data availability statement deposits the study's datasets (live-imaging/phenotyping data underlying the sepal growth analysis) in two public OSF repositories with explicit DOIs, making them paper-specific, public, and actionable.Dataset · publicg and Michelle Heeney for their comments on the manuscript. We thank Richard Smith (John Innes Centre, UK) for his advice on image processing.
Data availability statement
The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found below: https://doi.org/10.17605/OSF.IO/UMW9B , https://doi.org/10.17605/OSF.IO/P5Q39 .
Author contributions
AS: Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Validation, Visualization, Writing – original draft, Writing – review & editing. AR: Conceptualization, Data curation, Funding acquisition, Project administration, Supervision, WritinOpen asset ↗OSF · 10.17605/OSF.IO/UMW9Blines:234-260Dataset · publicon the manuscript. We thank Richard Smith (John Innes Centre, UK) for his advice on image processing.
Data availability statement
The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found below: https://doi.org/10.17605/OSF.IO/UMW9B , https://doi.org/10.17605/OSF.IO/P5Q39 .
Author contributions
AS: Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Validation, Visualization, Writing – original draft, Writing – review & editing. AR: Conceptualization, Data curation, Funding acquisition, Project administration, Supervision, Writing – review & editing.
Conflict of intereOpen asset ↗OSF · 10.17605/OSF.IO/P5Q39lines:234-260Code / dataset availability confirmedEurope PMC · checked 15 Sept 2026
Pollen is becoming an increasingly important subject for molecular researchers in genetic engineering, plant breeding, and environmental monitoring. To broaden the scope of these studies, it is essential to develop accessible methods for scientists who are not specialized in palynology. The article presents a simplified technical procedure for preparing pollen grains for scanning electron microscopy (SEM). The protocol is convenient for any molecular laboratory due to its small set of reagents, ease of execution, low cost, does not require special equipment, and takes only one hour to complete. The high penetrating ability of formaldehyde and the final delicate dehydration using hexamethyldisilazane (HMDS) instead of critical point drying allow for sufficient preservation of the architecture of the aperture, which is considered a gateway for the passage of biomolecules. The method was successfully applied to pollen grains of representatives of dicotyledons (beetroot, petunia, radish, tomato and tobacco) and monocotyledons (lily, onion, corn, rye and wheat). Species studied included insect-pollinated (entomophilous) and wind-pollinated (anemophilous) species. A comparative analysis of the sizes of fresh living pollen grains under a light microscope and those prepared for SEM showed some shrinkage. Quantitative analysis of the degree of pollen grain shrinkage showed that this process depends on the initial shape of dry pollen grains, and the number and structure of apertures. The results support the theoretical model of the folding/unfolding pathways of pollen grains.
Why it matches plant phenotyping methods植物花粉のSEM観察用試料調製法そのものを開発し、複数植物で適用・比較検証しているため、形態計測に関する中心的な方法論研究である。
abstractThe article presents a simplified technical procedure for preparing pollen grains for scanning electron microscopy (SEM).
Reproduction assets foundThe paper's quantitative pollen shrinkage measurements (Table S1) and light microscopy images (Figures S3–S4) are contained in the publicly downloadable MDPI Supplementary Materials, which directly reproduce this paper's phenotyping measurements. No author analysis code or trained models are mentioned.Supplement · publicoly Bogdanov—at the department of electron microscopy, Lomonosov Moscow State University.
Abbreviations
The following abbreviations are used in this manuscript:
SEM
Scanning Electron Microscopy
HMDS
Hexamethyldisilazane
SA
Short axis
LA
Long axis
Supplementary Materials
The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/plants13152140/s1 , Figure S1: The order of steps for pollen preparation according to the developed protocol; Figure S2: Scheme of measured pollen grain diameters; Figure S3: Light microscopy of pollen grains of insect-pollinated species; Figure S4: Light microscopy of pollen grains of wind-pollinated species; Table S1: Comparison Open asset ↗lines:98-127Code / dataset availability confirmedOpenAlex · Europe PMC · bioRxiv · checked 15 Sept 2026
Abstract Background Automating stomatal trait measurement has gained popularity because of their inherent importance for field phenotyping application as stomata are critical for both carbon capture and water use efficiency in plants. Such tool has been reported for rice, wheat, tomato, barley and oil palm. However, none exist yet for canola, which is an important economic and agronomic crop globally. Results We developed a new toolkit called Stomatal Comprehensive Automated Neural Network or SCAN by combining the use of high-resolution portable digital microscopy with machine learning based on You Only Look Once algorithm (YOLOv8). Digital micrographs of leaf surfaces enter the SCAN pipeline, which includes stomata detection, stomata segmentation and stomatal pore segmentation models, to measure stomatal density, stomatal size and stomatal pore area, respectively. In addition to SCAN’s ability to measure leaf stomatal traits in canola at 89 to 94% accuracy, we also showed that SCAN can be used to predict stomatal density even in species not included in the training set such as Arabidopsis, tobacco, rice, wheat, maize and proso millet. SCAN was designed for the biological science community with the premise that users are not required to possess advanced programming capabilities to manage dependency prerequisites, execute the models, and integrate the analysis. This was achieved by packaging the models into a desktop application system that can be accessed offline. Conclusion Overall, SCAN provides a non-destructive, real-time, portable, and high-throughput measurement of leaf stomatal traits in canola. The minimised hardware requirement and user-friendly desktop application system make SCAN suitable for field phenotyping application.
Why it matches plant phenotyping methodsカノーラ葉の気孔形質を画像と機械学習で自動抽出するツールを開発し、精度検証と他種での適用性評価を行っており、フェノタイピング手法が中心である。
abstractWe developed a new toolkit called Stomatal Comprehensive Automated Neural Network or SCAN by combining the use of high-resolution portable digital microscopy with machine learning based on You Only Look Once algorithm (YOLOv8).
Reproduction assets foundThe paper's authors publicly deposit the SCAN pipeline's model weights, hyperparameters, training scripts, and datasets in the FD_detection GitHub repository, and provide the SCAN application itself (with download and demonstration) in a second GitHub repository. Both are paper-specific, public, and actionable.Code · publicin Table S1.
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The training tasks were carried out on an Ubuntu 20.04 Linux server at the Research School of Biology in
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Australian National University, using two Nvidia A30 (24G) Graphic Processing Units (GPUs). The full
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details of models’ weights, hyperparameters, training scripts and datasets can be found at
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https://github.com/William-Yao0993/FD_detection.128
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Model evaluation
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Mean Average Precision (mAP, Fig. 3) and F1 score were used to assess model ability (Fig. 4). mAP is
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calculated as the mean value of each class area under the precision-recall curve over thresholds, and the
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F1 score is the harmonic mean of precision and recall. The formulas are deOpen asset ↗William-Yao0993/FD_detectionpdf-raw-page:4 lines:1-81Code / dataset availability confirmedCrossref · Europe PMC · checked 7 Sept 2026
Abstract Background The sweet potato whitefly ( Bemisia tabaci ) is a globally important insect pest that damages crops through direct feeding and by transmitting viruses. Current B. tabaci management revolves around the use of insecticides, which are economically and environmentally costly. Host plant resistance is a sustainable option to reduce the impact of whiteflies, but progress in deploying resistance in crops has been slow. A major obstacle is the high cost and low throughput of screening plants for B. tabaci resistance. Oviposition rate is a popular metric for host plant resistance to B. tabaci because it does not require tracking insect development through the entire life cycle, but accurate quantification is still limited by difficulties in observing B. tabaci eggs, which are microscopic and translucent. The goal of our study was to improve quantification of B. tabaci eggs on several important crop species: cassava, cowpea, melon, sweet potato and tomato. Results We tested a selective staining process originally developed for leafhopper eggs: submerging the leaves in McBryde’s stain (acetic acid, ethanol, 0.2% aqueous acid Fuchsin, water; 20:19:2:1) for three days, followed by clearing under heat and pressure for 15 min in clearing solution (LGW; lactic acid, glycerol, water; 17:20:23). With a less experienced individual counting the eggs, B. tabaci egg counts increased after staining across all five crops. With a more experienced counter, egg counts increased after staining on melons, tomatoes, and cowpeas. For all five crops, there was significantly greater agreement on egg counts across the two counting individuals after the staining process. The staining method worked particularly well on melon, where egg counts universally increased after staining for both counting individuals. Conclusions Selective staining aids visualization of B. tabaci eggs across multiple crop plants, particularly species where leaf morphological features obscure eggs, such as melons and tomatoes. This method is broadly applicable to research questions requiring accurate quantification of B. tabaci eggs, including phenotyping for B. tabaci resistance.
Why it matches plant phenotyping methods植物葉上のコナジラミ卵を染色して定量し、計数値と計数者間一致を改善する方法を評価しており、抵抗性フェノタイピングへの応用が明示された中心的な手法研究。
abstractThe goal of our study was to improve quantification of B. tabaci eggs on several important crop species: cassava, cowpea, melon, sweet potato and tomato.
Reproduction assets foundThe paper's Data availability statement explicitly deposits the study's egg-count datasets and the R Markdown analysis code in a Dryad repository, which is a public, paper-specific asset directly reproducing the phenotyping measurements and analysis.Dataset · publicThe datasets generated and analyzed during this study, and an R Markdown document containing the code used to perform these analyses are available in a Dryad repository (DOI: doi: https://doi.org/10.5061/dryad.vmcvdnd1m ).Open asset ↗Dryad · 10.5061/dryad.vmcvdnd1mlines:138-163Code / dataset availability confirmedEurope PMC · checked 7 Sept 2026
Cell plate formation during cytokinesis entails multiple stages occurring concurrently and requiring orchestrated vesicle delivery, membrane remodelling, and timely deposition of polysaccharides, such as callose. Understanding such a dynamic process requires dissection in time and space; this has been a major hurdle in studying cytokinesis. Using lattice light sheet microscopy (LLSM), we studied cell plate development in four dimensions, through the behavior of yellow fluorescent protein (YFP)-tagged cytokinesis-specific GTPase RABA2a vesicles. We monitored the entire duration of cell plate development, from its first emergence, with the aid of YFP-RABA2a, in both the presence and absence of cytokinetic callose. By developing a robust cytokinetic vesicle volume analysis pipeline, we identified distinct behavioral patterns, allowing the identification of three easily trackable cell plate developmental phases. Notably, the phase transition between phase I and phase II is striking, indicating a switch from membrane accumulation to the recycling of excess membrane material. We interrogated the role of callose using pharmacological inhibition with LLSM and electron microscopy. Loss of callose inhibited the phase transitions, establishing the critical role and timing of the polysaccharide deposition in cell plate expansion and maturation. This study exemplifies the power of combining LLSM with quantitative analysis to decode and untangle such a complex process.
Why it matches plant phenotyping methodsLLSMによる4次元画像取得と、細胞板の小胞体積を定量化する解析パイプラインの開発が研究の中心であり、植物細胞の形態・発達状態を抽出する方法として substantive です。
abstractUsing lattice light sheet microscopy (LLSM), we studied cell plate development in four dimensions
Reproduction assets foundThe paper deposits representative 4D lattice light sheet microscopy datasets (YFP–RABA2a cell plate imaging) used for its quantitative analysis on Zenodo, a paper-specific public asset. No author analysis code repository with explicit availability language is stated; the other URLs are method guidelines, not paper dataDataset · publicRepresentative datasets used in the study are available on Zenodo at https://doi.org/10.5281/zenodo.10515765 .Open asset ↗Zenodo · 10.5281/zenodo.10515765lines:85-93Code / dataset availability confirmedEurope PMC · bioRxiv · checked 15 Sept 2026
Summary StomaVision is an automated tool designed for high-throughput detection and measurement of stomatal traits, such as stomatal number, pore size, and closure rate. It provides insights into plant responses to environmental cues, streamlining the analysis of micrographs from field-grown plants across various species, including monocots and dicots. Enhanced by a novel collection method that utilizes video recording, StomaVision increases the number of captured images for robust statistical analysis. Accessible via an intuitive web interface at and available for local use in a containerized environment at , this tool ensures long-term usability by minimizing the impact of software updates and maintaining functionality with minimal setup requirements. The application of StomaVision has provided significant physiological insights, such as variations in stomatal density, opening rates, and total pore area under heat stress. These traits correlate with critical physiological processes, including gas exchange, carbon assimilation, and water use efficiency, demonstrating the tool’s utility in advancing our understanding of plant physiology. The ability of StomaVision to identify differences in responses to varying durations of heat treatment highlights its value in plant science research. Plain language summary StomaVision is a tool that automatically counts and measures tiny openings on plant leaves, helping us learn how plants deal with their surroundings. It is easy to use and works well with various plant species. This tool helps scientists see how plants change under stress, making plant research easier and more accurate.
Why it matches plant phenotyping methods気孔数、孔サイズ、閉鎖率などの植物形質を画像から自動抽出するツールの開発・提供が研究の中心であり、植物フェノタイピング手法に該当する。
abstractStomaVision is an automated tool designed for high-throughput detection and measurement of stomatal traits, such as stomatal number, pore size, and closure rate.
Reproduction assets foundThe authors publicly release their StomaVision source code, trained YOLOv7-seg model, and all labeled stomata images on GitHub, plus a public Streamlit web portal for stomatal trait analysis. Cited datasets (Dryad/LeafNet, Cuticle Database) and generic libraries (VDP, Detectron2, Ultralytics, Label Studio) are prior/thCode · publicl for advancing our understanding of stomatal behavior,
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particularly in an era in which plant resilience and adaptation are of paramount
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concern.
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Data Availability
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The source code, trained model, user installation and training guideline, and all the
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labeled images of leaf stomata are available at
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https://github.com/YaoChengLab/StomaVision. The web portal of extracting stomatal
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traits is available at https://stomavision.streamlit.app/.850
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Author Contributions
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TLW, PYC, XD, PLC, and YCL conceived the research. TLW, JYO, PXZ, YLW, RHW,
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TCH, CYL, and YCL conducted the field and growth chamber experiments. TLW,
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JYO, PXZ, YLW, and RHW produceOpen asset ↗YaoChengLab/StomaVisionpdf-raw-page:27 lines:1-65Code / dataset availability confirmedEurope PMC · checked 15 Sept 2026
The periderm is a vital protective tissue found in the roots, stems, and woody elements of diverse plant species. It plays an important function in these plants by assuming the role of the epidermis as the outermost layer. Despite its critical role for protecting plants from environmental stresses and pathogens, research on root periderm development has been limited due to its late formation during root development, its presence only in mature root regions, and its impermeability. One of the most straightforward measurements for comparing periderm formation between different genotypes and treatments is periderm (phellem) length. We have developed PAT (Periderm Assessment Toolkit), a high-throughput user-friendly pipeline that integrates an efficient staining protocol, automated imaging, and a deep-learning-based image analysis approach to accurately detect and measure periderm length in the roots of Arabidopsis thaliana . The reliability and reproducibility of our method was evaluated using a diverse set of 20 Arabidopsis natural accessions. Our automated measurements exhibited a strong correlation with human-expert-generated measurements, achieving a 94% efficiency in periderm length quantification. This robust PAT pipeline streamlines large-scale periderm measurements, thereby being able to facilitate comprehensive genetic studies and screens. Although PAT proves highly effective with automated digital microscopes in Arabidopsis roots, its application may pose challenges with nonautomated microscopy. Although the workflow and principles could be adapted for other plant species, additional optimization would be necessary. While we show that periderm length can be used to distinguish a mutant impaired in periderm development from wild type, we also find it is a plastic trait. Therefore, care must be taken to include sufficient repeats and controls, to minimize variation, and to ensure comparability of periderm length measurements between different genotypes and growth conditions.
Why it matches plant phenotyping methods植物根の表現型（周皮長）を自動画像取得・深層学習解析で定量するパイプラインを開発し、専門家測定との相関で信頼性と再現性を検証しており、方法が研究の中心である。
abstractWe have developed PAT (Periderm Assessment Toolkit), a high-throughput user-friendly pipeline that integrates an efficient staining protocol, automated imaging, and a deep-learning-based image analysis approach to accurately detect and measure periderm length in the roots of Arabidopsis thaliana .
Reproduction assets foundThe authors publicly release the PAT pipeline (analysis code/scripts) and a test dataset of Col-0 and wox4-1 TIFF microscopy images via their GitHub repository. Full-resolution TIFF images of the 20 natural accessions are only available upon request (request_only, not listed as an allowed URL).Dataset · publicroved the manuscript.
Competing interests: W.B. is a cofounder of Cquesta, a company that works on crop root growth and carbon sequestration.
Data Availability
All raw data and datasets have been included in the Supplementary Materials. The PAT pipeline and its associated code are accessible via the following GitHub repository: https://github.com/Salk-Harnessing-Plants-Initiative/PAT-Pipeline-for-Analysis-of-Periderm . Additionally, the test dataset comprising Col-0 and wox4-1 TIFF images is available on the same GitHub repository. Full-resolution TIFF images corresponding to the natural accessions (Table 1 ) can be obtained from the corresponding author upon request.
Supplementary MaterialsOpen asset ↗Salk-Harnessing-Plants-Initiative/PAT-Pipeline-for-Analysis-of-Peridermlines:391-421Code / dataset availability confirmedEurope PMC · checked 15 Sept 2026
Correlative light and electron microscopy (CLEM) is an important tool for the localisation of target molecule(s) and their spatial correlation with the ultrastructural map of subcellular features at the nanometre scale. Adoption of these advanced imaging methods has been limited in plant biology, due to challenges with plant tissue permeability, fluorescence labelling efficiency, indexing of features of interest throughout the complex 3D volume and their re-localization on micrographs of ultrathin cross-sections. Here, we demonstrate an imaging approach based on tissue processing and embedding into methacrylate resin followed by imaging of sections by both, single-molecule localization microscopy and transmission electron microscopy using consecutive CLEM and same-section CLEM correlative workflow. Importantly, we demonstrate that the use of a particular type of embedding resin is not only compatible with single-molecule localization microscopy but shows improvements in the fluorophore blinking behavior relative to the whole-mount approaches. Here, we use a commercially available Click-iT ethynyl-deoxyuridine cell proliferation kit to visualize the DNA replication sites of wild-type Arabidopsis thaliana seedlings, as well as fasciata1 and nucleolin1 plants and apply our in-section CLEM imaging workflow for the analysis of S-phase progression and nucleolar organization in mutant plants with aberrant nucleolar phenotypes.
Why it matches plant phenotyping methods植物組織に適用するin-section CLEMおよび超解像イメージングのワークフローを開発・実証しており、植物細胞の構造・複製状態を取得する方法が研究の中心である。
abstractHere, we demonstrate an imaging approach based on tissue processing and embedding into methacrylate resin followed by imaging of sections by both, single-molecule localization microscopy and transmission electron microscopy using consecutive CLEM and same-section CLEM correlative workflow.
Reproduction assets foundThe paper's raw TEM and SMLM phenotyping image datasets (Arabidopsis nucleolar ultrastructure and DNA replication CLEM analysis) are publicly deposited in the BioImage Archive under accession S-BIAD700, with an explicit authors' URL. Source data quantification sheets are only provided with the paper, not as a separate址Dataset · publicature Communications thanks the anonymous reviewers for their contribution to the peer review of this work. A peer review file is available.
Data availability
The raw datasets of TEM imaging (Spurr and Lowicryl) and SMLM data for quantitative analysis have been deposited to the BioImage Archive, under accession code S-BIAD700 ( https://www.ebi.ac.uk/biostudies/bioimages/studies/S-BIAD700 ). Source data are provided with this paper (sheet 1—IRF quantification, sheet 2—FC quantification, sheet 3—DBSCAN analysis). Source data are provided with this paper.
Competing interests
The authors declare no competing interests.
Footnotes
Publisher’s note Springer Nature remains neutral with regard to jurOpen asset ↗BioImage Archive · S-BIAD700lines:116-150Code / dataset availability confirmedEurope PMC · checked 7 Sept 2026
Introduction Plants undergo various natural changes that dramatically modify their genomes. One is polyploidization and the second is hybridization. Both are regarded as key factors in plant evolution and result in phenotypic differences in different plant organs. In Silene , we can find both examples in nature, and this genus has a seed shape diversity that has long been recognized as a valuable source of information for infrageneric classification. Methods Morphometric analysis is a statistical study of shape and size and their covariations with other variables. Traditionally, seed shape description was limited to an approximate comparison with geometric figures (rounded, globular, reniform, or heart-shaped). Seed shape quantification has been based on direct measurements, such as area, perimeter, length, and width, narrowing statistical analysis. We used seed images and processed them to obtain silhouettes. We performed geometric morphometric analyses, such as similarity to geometric models and elliptic Fourier analysis, to study the hybrid offspring of S. latifolia and S. dioica . Results We generated synthetic tetraploids of Silene latifolia and performed controlled crosses between diploid S. latifolia and Silene dioica to analyze seed morphology. After imaging capture and post-processing, statistical analysis revealed differences in seed size, but not in shape, between S. latifolia diploids and tetraploids, as well as some differences in shape among the parentals and hybrids. A detailed inspection using fluorescence microscopy allowed for the identification of shape differences in the cells of the seed coat. In the case of hybrids, differences were found in circularity and solidity. Overal seed shape is maternally regulated for both species, whereas cell shape cannot be associated with any of the sexes. Discussion Our results provide additional tools useful for the combination of morphology with genetics, ecology or taxonomy. Seed shape is a robust indicator that can be used as a complementary tool for the genetic and phylogenetic analyses of Silene hybrid populations.
Why it matches plant phenotyping methods種子画像を処理し、幾何学的形態計測と楕円フーリエ解析で種子形状・サイズを定量化する手法が研究の中心であり、植物器官の形態表現型を抽出している。
abstractWe used seed images and processed them to obtain silhouettes. We performed geometric morphometric analyses, such as similarity to geometric models and elliptic Fourier analysis, to study the hybrid offspring of S. latifolia and S. dioica .
Reproduction assets foundThe authors state that the raw seed images used for the morphometric phenotyping analyses are publicly deposited in Zenodo (DOI 10.5281/zenodo.8366177). This is a paper-specific, publicly accessible dataset of the seed/cell images underlying this study's measurements. No author analysis code with an explicit public URLDataset · publicRaw images used in this work are available in Zenodo DOI 10.5281/zenodo.8366177 .Open asset ↗Zenodo · 10.5281/zenodo.8366177lines:436-491Code / dataset availability confirmedbioRxiv · checked 15 Sept 2026
Variation in Drosophila compound eye size is studied across research fields, from evolutionary biology to biomedical studies, requiring the collection of large datasets to ensure robust statistical analyses. To address this, we present EyeHex, a tool for automatic segmentation of fruit fly compound eyes from brightfield and scanning electron microscopy (SEM) images. EyeHex features two integrated modules: the first utilizes machine learning to generate probability maps of the eye and ommatidia locations, while the second, a hard-coded module, leverages the hexagonal organization of the compound eye to map individual ommatidia. This iterative segmentation process, which adds one ommatidium at a time based on registered neighbors, ensures robustness to local perturbations. EyeHex also includes an analysis tool that calculates key metrics of the eye, such as ommatidia count and diameter distribution across the eye. With minimal user input for training and application, EyeHex achieves exceptional accuracy (>99.6% compared to manual counts on SEM images) and adapts to different fly strains, species, and image types. EyeHex offers a cost-effective, rapid, and flexible pipeline for extracting detailed statistical data on Drosophila compound eye variation, making it a valuable resource for high-throughput studies.
Why it matches plant phenotyping methods昆虫（ショウジョウバエ）の眼を対象としており植物ではないため、植物フェノタイピング文献の対象外です。
abstractautomatic segmentation of fruit fly compound eyes from brightfield and scanning electron microscopy (SEM) images
Reproduction assets foundThe paper's EyeHex MATLAB toolbox (with manual and sample images) and the post-segmentation analysis code are publicly available on GitHub, as stated in the Declarations. The segmentation results/analysis supplement is only a PDF supplement; the toolbox and analysis code are the paper-specific public assets.Code · publict of abbreviations
A-P: Anterior-Posterior
CT: Micro Computed Tomography
SEM: Scanning Electron Microscopy
GUI: Graphical User Interface
Declarations
Ethics approval and consent to participate
Not applicable
Consent for publication
Not applicable
Availability of data and Supplementary materials
EyeHex toolbox is available from https://github.com/huytran216/EyeHex-toolbox.
The analysis code following EyeHex segmentation for all eyes in the dataset can be
downloaded from https://github.com/huytran216/EyeHex_analysis.
Segmentation results and analysis for Hikone-AS (26 eyes) and Canton-SBH (12 eyes):
Supplementary_file.pdf
Competing interests
The authors declare that they have no competing intOpen asset ↗huytran216/EyeHex-toolboxpdf-layout-page:22 lines:1-53Code · publictions
Ethics approval and consent to participate
Not applicable
Consent for publication
Not applicable
Availability of data and Supplementary materials
EyeHex toolbox is available from https://github.com/huytran216/EyeHex-toolbox.
The analysis code following EyeHex segmentation for all eyes in the dataset can be
downloaded from https://github.com/huytran216/EyeHex_analysis.
Segmentation results and analysis for Hikone-AS (26 eyes) and Canton-SBH (12 eyes):
Supplementary_file.pdf
Competing interests
The authors declare that they have no competing interests.
Author’s contributions
AR collected the data (sample preparation and imaging), HT created EyeHex toolbox
and analyzed the data, AR and HTOpen asset ↗huytran216/EyeHex_analysispdf-layout-page:22 lines:1-53Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
Published4 Feb 2024The Plant journal : for cell and molecular biologyCited by 0 · OpenAlex ↗
Confocal microscopy has greatly aided our understanding of the major cellular processes and trafficking pathways responsible for plant growth and development. However, a drawback of these studies is that they often rely on the manual analysis of a vast number of images, which is time-consuming, error-prone, and subject to bias. To overcome these limitations, we developed Dot Scanner, a Python program for analyzing the densities, lifetimes, and displacements of fluorescently tagged particles in an unbiased, automated, and efficient manner. Dot Scanner was validated by performing side-by-side analysis in Fiji-ImageJ of particles involved in cellulose biosynthesis. We found that the particle densities and lifetimes were comparable in both Dot Scanner and Fiji-ImageJ, verifying the accuracy of Dot Scanner. Dot Scanner largely outperforms Fiji-ImageJ, since it suffers far less selection bias when calculating particle lifetimes and is much more efficient at distinguishing between weak signals and background signal caused by bleaching. Not only does Dot Scanner obtain much more robust results, but it is a highly efficient program, since it automates much of the analyses, shortening workflow durations from weeks to minutes. This free and accessible program will be a highly advantageous tool for analyzing live-cell imaging in plants.
Why it matches plant phenotyping methods植物のライブセル画像から粒子密度・寿命・変位を自動抽出するソフトウェアを開発し、Fiji-ImageJと比較検証しており、表現型取得・解析手法が中心である。
abstractwe developed Dot Scanner, a Python program for analyzing the densities, lifetimes, and displacements of fluorescently tagged particles
Reproduction assets foundThe paper's own computational analysis tool, Dot Scanner (Python software for quantifying densities, lifetimes, and displacements of fluorescently labeled particles in plant tissues), is explicitly stated to be publicly available on GitHub with a full URL. No public phenotype/trait datasets or raw imaging data depositsCode · public= 2, blob size = 5, dot lower = 0.9, dot
upper = 4.5, and blob lower = 2, skips = 1, and remove edge
frames = false. All lifetimes that were 60 sec long were removed
from the analysis.
Dot scanner
Dot Scanner was developed using the Python programming lan-
guage. The software is available on GitHub, and the project home-
page (https://github.com/bdavis222/dotscanner) contains all the
documentation needed for its installation and use, including
the README file (https://github.com/bdavis222/dot-scanner/blob/main/README.md). As mentioned in the README, Python 3 must
be installed prior to Dot Scanner installation (https://www.python.org/downloads/).ACKNOWLEDGMENTS
We thank S. Bednarek for provOpen asset ↗bdavis222/dotscannerpdf-raw-page:9 lines:1-87Code / dataset availability confirmedEurope PMC · bioRxiv · checked 14 Sept 2026
ArabidopsisMesh / voxelMicroscopyFlowerSegmentationGrowth / time-series analysisTrackingGrowth / development / phenology
Background Arabidopsis thaliana sepals are excellent models for analyzing growth of entire organs due to their relatively small size, which can be captured at a cellular resolution under a confocal microscope [1]. To investigate how growth of different tissue layers generates unique organ morphologies, it is necessary to live-image deep into the tissue. However, imaging deep cell layers of the sepal is practically challenging, as it is hindered by the presence of extracellular air spaces between mesophyll cells, among other factors which causes optical aberrations. Image processing is also difficult due to the low signal-to-noise ratio of the deeper tissue layers, an issue mainly associated with live imaging datasets. Addressing some of these challenges, we provide an optimized methodology for live imaging sepals and subsequent image processing. This helps us track the growth of individual cells on the outer and inner epidermal layers, which are the key drivers of sepal morphogenesis. Results For live imaging sepals across all tissue layers at early stages of development, we found that the use of a bright fluorescent membrane marker, coupled with increased laser intensity and an enhanced Z-resolution produces high-quality images suitable for downstream image processing. Our optimized parameters allowed us to image the bottommost cell layer of the sepal (inner epidermal layer) without compromising viability. We used a ‘voxel removal’ technique to visualize the inner epidermal layer in MorphoGraphX [2, 3] image processing software. Finally, we describe the process of optimizing the parameters for creating a 2.5D mesh surface for the inner epidermis. This allowed segmentation and parent tracking of individual cells through multiple time points, despite the weak signal of the inner epidermal cells. Conclusion We provide a robust pipeline for imaging and analyzing growth across inner and outer epidermal layers during early sepal development. Our approach can potentially be employed for analyzing growth of other internal cell layers of the sepals as well. For each of the steps, approaches, and parameters we used, we have provided in-depth explanations to help researchers understand the rationale and replicate our pipeline.
Why it matches plant phenotyping methodsライブイメージング、画像処理、細胞セグメンテーションと追跡を統合した、萼片の成長・形態解析パイプラインの最適化が中心であり、植物表現型取得法に該当する。
abstractwe provide an optimized methodology for live imaging sepals and subsequent image processing
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Dataset · publicThe images of the WT flowers, as well as the final edited images can be accessed at https://doi.org/10.17605/OSF.IO/UMW9B . The images shown in this manuscript correspond to WT replicate 2.Open asset ↗OSF.IO/UMW9Blines:137-166Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
Published24 Jan 2024The Plant journal : for cell and molecular biologyCited by 8 · OpenAlex ↗
The model moss species Physcomitrium patens has long been used for studying divergence of land plants spanning from bryophytes to angiosperms. In addition to its phylogenetic relationships, the limited number of differential tissues, and comparable morphology to the earliest embryophytes provide a system to represent basic plant architecture. Based on plant-fungal interactions today, it is hypothesized these kingdoms have a long-standing relationship, predating plant terrestrialization. Mortierellaceae have origins diverging from other land fungi paralleling bryophyte divergence, are related to arbuscular mycorrhizal fungi but are free-living, observed to interact with plants, and can be found in moss microbiomes globally. Due to their parallel origins, we assess here how two Mortierellaceae species, Linnemannia elongata and Benniella erionia, interact with P. patens in coculture. We also assess how Mollicute-related or Burkholderia-related endobacterial symbionts (MRE or BRE) of these fungi impact plant response. Coculture interactions are investigated through high-throughput phenomics, microscopy, RNA-sequencing, differential expression profiling, gene ontology enrichment, and comparisons among 99 other P. patens transcriptomic studies. Here we present new high-throughput approaches for measuring P. patens growth, identify novel expression of over 800 genes that are not expressed on traditional agar media, identify subtle interactions between P. patens and Mortierellaceae, and observe changes to plant-fungal interactions dependent on whether MRE or BRE are present. Our study provides insights into how plants and fungal partners may have interacted based on their communications observed today as well as identifying L. elongata and B. erionia as modern fungal endophytes with P. patens.
Why it matches plant phenotyping methodsP. patensの成長を測定する新しい高スループット手法とフェノミクス解析を提示しており、植物表現型取得が研究の実質的な方法的貢献である。
abstractCoculture interactions are investigated through high-throughput phenomics, microscopy, RNA-sequencing, differential expression profiling, gene ontology enrichment, and comparisons among 99 other P. patens transcriptomic studies.
Reproduction assets foundThe paper deposits authors' supplementary analysis code (DESeq2 differential expression and comparison scripts) on Zenodo and supplementary data on Dryad, both with explicit availability statements and public URLs. The paper also describes Raspberry Pi/ArduCam/PlantCV imaging hardware and software for phenotyping, but Dataset · publices. In particular, the University resides on Land DATA AVAILABILITY STATEMENT
ceded in the 1819 Treaty of Saginaw. We recognize, support, and
advocate for the sovereignty of Michigan’s 12 federally recognized The following Supplementary Data have been deposited at
Indian nations, for historic Indigenous communities in Michigan, https://datadryad.org/stash/share/2g3gZefPksJaPGlLpc8d7g
Ó 2024 The Authors.
The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.,
The Plant Journal, (2024), doi: 10.1111/tpj.16605Open asset ↗datadryad · 2g3gZefPksJaPGlLpc8d7gpdf-layout-page:16 lines:58-78Code / dataset availability confirmedCrossref · checked 15 Sept 2026
Quantitative phase imaging by digital holographic microscopy (DHM) is a nondestructive and label-free technique that has been playing an indispensable role in the fields of science, technology, and biomedical imaging. The technique is competent in imaging and analyzing label-free living cells and investigating reflective surfaces. Herein, we introduce a new configuration of a wide field-of-view single-shot common-path off-axis reflective DHM for the quantitative phase imaging of biological cells that leverages several advantages, including being less-vibration sensitive to external perturbations due to its common-path configuration, also being compact in size, simple in optical design, highly stable, and cost-effective. A detailed description of the proposed DHM system, including its optical design, working principle, and capability for phase imaging, is presented. The applications of the proposed system are demonstrated through quantitative phase imaging results obtained from the reflective surface (USAF resolution test target) as well as transparent samples (living plant cells). The proposed system could find its applications in the investigation of several biological specimens and the optical metrology of micro-surfaces.
Why it matches plant phenotyping methods植物細胞を対象に定量位相画像を取得できる新規デジタルホログラフィック顕微鏡を開発しており、植物試料での性能実証も含むため、植物フェノタイピングに利用可能な画像計測法が中心です。
abstractwe introduce a new configuration of a wide field-of-view single-shot common-path off-axis reflective DHM for the quantitative phase imaging of biological cells
Reproduction assets foundThe paper's only paper-specific public asset is the MDPI supplementary material containing the time-lapse retrieved wrapped phase imaging video of tobacco plant cells (the paper's plant-cell phenotyping measurements). No analysis code, datasets, or trained models are publicly deposited; the Data Availability Statement仅Supplement · publicdual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.
Supplementary Materials
The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/s24030720/s1 , See the supplementary material for visualization of the time-lapse retrieved wrapped phase imaging video of tobacco plant cells captured five-minute intervals.
Click here for additional data file.
Author Contributions
Conceptualization: M.K.; methodology, M.K.; software, M.K.; validation, M.K., O.M. and T.M.; formal Open asset ↗lines:51-66Code / dataset availability confirmedEurope PMC · Crossref · bioRxiv · checked 15 Sept 2026
Chloroplasts are essential organelles in plants that are involved in plant development and photosynthesis. Accurate quantification of chloroplast numbers is important for understanding the status and type of plant cells, as well as assessing photosynthetic potential and efficiency. Traditional methods of counting chloroplasts using microscopy are time-consuming and face challenges such as the possibility of missing out-of-focus samples or double counting when adjusting the focal position. Here, we developed an innovative approach called Detecting- and-Counting-chloroplasts (D&Cchl) for automated detection and counting of chloroplasts. This approach utilizes a deep-learning-based object detection algorithm called You-Only-Look-Once (YOLO), along with the Intersection Over Union (IOU) strategy. The application of D&Cchl has shown excellent performance in accurately identifying and quantifying chloroplasts. This holds true when applied to both a single image and a three-dimensional (3D) structure composed of a series of images. Furthermore, by integrating Cellpose, a cell-segmentation tool, we were able to successfully perform single-cell 3D chloroplast counting. Compared to manual counting methods, this approach improved the accuracy of detection and counting to over 95%. Together, our work not only provides an efficient and reliable tool for accurately analyzing the status of chloroplasts, enhancing our understanding of plant photosynthetic cells and growth characteristics, but also makes a significant contribution to the convergence of botany and deep learning. One-sentence summary This deep learning-based approach enables the accurate complete detection and counting of chloroplasts in 3D single cells using microscopic image stacks, and showcases a successful example of utilizing deep learning methods to analyze subcellular spatial information in plant cells. The authors responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors ( https://academic.oup.com/plcell/ ) is: Zhao Dong ( dongzhao@hebeu.edu.cn ), Shaokai Yang, ( shaokai1@ualberta.ca ), Ningjing Liu ( liuningjing1@yeah.net ), and Qiong Zhao ( qzhao@bio.ecnu.edu.cn ).
Why it matches plant phenotyping methods植物細胞の顕微鏡画像から葉緑体数を自動検出・定量する深層学習手法を開発し、手動計数と比較して精度検証しているため、植物フェノタイピング手法が中心である。
abstractHere, we developed an innovative approach called Detecting- and-Counting-chloroplasts (D&Cchl) for automated detection and counting of chloroplasts.
Reproduction assets foundThe authors explicitly state that all code and the training dataset (annotated chloroplast microscopy images) are shared on their public GitHub repository, which is a paper-specific asset for this chloroplast counting study. Other URLs (labelImg, yolov7, ImageJ Falk plugins) are generic third-party tools, not paper-ownCode · publicltiple times during the stacking process. Through this approach, we
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successfully constructed a comprehensive 3D cell model from the series of 2D
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images, enabling more accurate chloroplast detection and counting in a 3D space.
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Code and software
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All the code and training dataset have been shared on GitHub
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(https://github.com/xiaoli111111111/-AI4CELLBIO-ECNU), with detailed
466
explanations in the supplementary manual.
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References
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preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this
this version posted December 18, 2023.
;
https://doi.org/10.1101/2023.12.Open asset ↗xiaoli111111111/-AI4CELLBIO-ECNUpdf-raw-page:16 lines:1-65Code / dataset availability confirmedEurope PMC · Crossref · checked 7 Sept 2026
Background Spatial mapping of transcriptional states provides valuable biological insights into cellular functions and interactions in the context of the tissue. Accurate 3D cell segmentation is a critical step in the analysis of this data towards understanding diseases and normal development in situ. Current approaches designed to automate 3D segmentation include stitching masks along one dimension, training a 3D neural network architecture from scratch, and reconstructing a 3D volume from 2D segmentations on all dimensions. However, the applicability of existing methods is hampered by inaccurate segmentations along the non-stitching dimensions, the lack of high-quality diverse 3D training data, and inhomogeneity of image resolution along orthogonal directions due to acquisition constraints; as a result, they have not been widely used in practice. Methods To address these challenges, we formulate the problem of finding cell correspondence across layers with a novel optimal transport (OT) approach. We propose CellStitch, a flexible pipeline that segments cells from 3D images without requiring large amounts of 3D training data. We further extend our method to interpolate internal slices from highly anisotropic cell images to recover isotropic cell morphology. Results We evaluated the performance of CellStitch through eight 3D plant microscopic datasets with diverse anisotropic levels and cell shapes. CellStitch substantially outperforms the state-of-the art methods on anisotropic images, and achieves comparable segmentation quality against competing methods in isotropic setting. We benchmarked and reported 3D segmentation results of all the methods with instance-level precision, recall and average precision (AP) metrics. Conclusions The proposed OT-based 3D segmentation pipeline outperformed the existing state-of-the-art methods on different datasets with nonzero anisotropy, providing high fidelity recovery of 3D cell morphology from microscopic images.
Why it matches plant phenotyping methods植物の3D顕微鏡画像から細胞形態を抽出するセグメンテーション手法を開発し、植物データセットで性能評価・ベンチマークしているため、植物フェノタイピング手法が中心である。
abstractWe propose CellStitch, a flexible pipeline that segments cells from 3D images without requiring large amounts of 3D training data.
Reproduction assets foundThe paper provides public author code (CellStitch implementation and experiment-reproducing notebooks on GitHub) and the plant image datasets analyzed (Ovules, ATAS, Arabidopsis 3D Digital Tissue Atlas), all with explicit availability statements and URLs.Code · publicopen source code implementing the stitching algorithm from the top to the bottom layer is available at https://github.com/imyiningliu/cellstitchOpen asset ↗imyiningliu/cellstitchlines:116-127Dataset · publicAll the datasets analyzed in this paper are publicly available online. Ovules: https://osf.io/uzq3w/ ; ATAS: https://www.repository.cam.ac.uk/handle/1810/262530 ; Arabidopsis 3D Digital Tissue Atlas: https://osf.io/fzr56Open asset ↗lines:175-223Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
The quantification of stomatal pore size has long been a fundamental approach to understand the physiological response of plants in the context of environmental adaptation. Automation of such methodologies not only alleviates human labor and bias but also realizes new experimental research methods through massive analysis. Here, we present an image analysis pipeline that automatically quantifies stomatal aperture of Arabidopsis thaliana leaves from bright-field microscopy images containing mesophyll tissue as noisy backgrounds. By combining a You Only Look Once X-based stomatal detection submodule and a U-Net-based pore segmentation submodule, we achieved a mean average precision with an intersection of union (IoU) threshold of 50% value of 0.875 (stomata detection performance) and an IoU of 0.745 (pore segmentation performance) against images of leaf discs taken with a bright-field microscope. Moreover, we designed a portable imaging device that allows easy acquisition of stomatal images from detached/undetached intact leaves on-site. We demonstrated that this device in combination with fine-tuned models of the pipeline we generated here provides robust measurements that can substitute for manual measurement of stomatal responses against pathogen inoculation. Utilization of our hardware and pipeline for automated stomatal aperture measurements is expected to accelerate research on stomatal biology of model dicots.
Why it matches plant phenotyping methods葉画像から気孔開度を自動抽出する画像解析パイプラインと携帯型撮像装置を開発・性能評価しており、植物表現型取得が中心である。
abstractwe present an image analysis pipeline that automatically quantifies stomatal aperture of Arabidopsis thaliana leaves
Reproduction assets foundThe paper's Data Availability section explicitly deposits the authors' ONNX model weights and pipeline code, plus masked/unmasked test images, on a public GitHub repository and Zenodo (DOI 10.5281/zenodo.7549843). These are paper-specific, publicly actionable assets for the stomatal aperture phenotyping pipeline. The YCode · publicPlant Cell Physiol. 00(00): 1–10 (2023) doi:https://doi.org/10.1093/pcp/pcad018
Supplementary Data
Supplementary data are available at PCP online.
Data Availability
The model weights and codes in ONNX format to execute
the Arabidopsis stomata quantification pipeline and mask
and unmasked test data are available at https://github.com/phytometrics/arabidopsis_leaf_stomata_quantification.Model weights and the test images are also available on Zen-
odo with the following DOI: https://doi.org/10.5281/zenodo.7549843. The software used to process image streams from the
portable device is under development at https://github.com/phytometrics/cvgui_linux.Funding
Grant-in-Aid for Transformative ResearcOpen asset ↗phytometrics/arabidopsis_leaf_stomata_quantificationpdf-raw-page:9 lines:1-84Dataset · publicData Availability
The model weights and codes in ONNX format to execute
the Arabidopsis stomata quantification pipeline and mask
and unmasked test data are available at https://github.com/phytometrics/arabidopsis_leaf_stomata_quantification.Model weights and the test images are also available on Zen-
odo with the following DOI: https://doi.org/10.5281/zenodo.7549843. The software used to process image streams from the
portable device is under development at https://github.com/phytometrics/cvgui_linux.Funding
Grant-in-Aid for Transformative Research Areas (21H05151 and
21H05149 to A.M. and 21H05152 to Y.T.), Grant-in-Aid for Sci-
entific Research (B) (19H02960 to A. M.), and Grant-in-Aid foOpen asset ↗10.5281/zenodo.7549843pdf-raw-page:9 lines:1-84Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
One of the fundamental questions in plant developmental biology is how cell proliferation and cell expansion coordinately determine organ growth and morphology. An amenable system to address this question is the Arabidopsis root tip, where cell proliferation and elongation occur in spatially separated domains, and cell morphologies can easily be observed using a confocal microscope. While past studies revealed numerous elements of root growth regulation including gene regulatory networks, hormone transport and signaling, cell mechanics and environmental perception, how cells divide and elongate under possible constraints from cell lineages and neighboring cell files has not been analyzed quantitatively. This is mainly due to the technical difficulties in capturing cell division and elongation dynamics at the tip of growing roots, as well as an extremely labor-intensive task of tracing the lineages of frequently dividing cells. Here, we developed a motion-tracking confocal microscope and an Artificial Intelligence (AI)-assisted image-processing pipeline that enables semi-automated quantification of cell division and elongation dynamics at the tip of vertically growing Arabidopsis roots. We also implemented a data sonification tool that facilitates human recognition of cell division synchrony. Using these tools, we revealed previously unnoted lineage-constrained dynamics of cell division and elongation, and their contribution to the root zonation boundaries.
Why it matches plant phenotyping methods生長中のシロイヌナズナ根端における細胞分裂・伸長という植物形態動態を、4D顕微鏡、AI画像処理、追跡、データソニフィケーションで半自動定量する手法を開発しており、フェノタイピング手法が中心である。
abstractHere, we developed a motion-tracking confocal microscope and an Artificial Intelligence (AI)-assisted image-processing pipeline that enables semi-automated quantification of cell division and elongation dynamics at the tip of vertically growing Arabidopsis roots.
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Code · publicThe codes for the nuclei detection and cell tracking are available on the GitHub ( https://github.com/JerrySongCST/Arabidopsis_root_cortex_cell_tracking ).Open asset ↗JerrySongCST/Arabidopsis_root_cortex_cell_trackinglines:152-227Code / dataset availability confirmedEurope PMC · checked 7 Sept 2026
In vitro pollen germination is considered the most efficient method to assess pollen viability. The pollen germination frequency and pollen tube length, which are key indicators of pollen viability, should be accurately measured during in vitro culture. In this study, a Mask R-CNN model trained using microscopic images of tree peony (Paeonia suffruticosa) pollen has been proposed to rapidly detect the pollen germination rate and pollen tube length. To reduce the workload during image acquisition, images of synthesized crossed pollen tubes were added to the training dataset, significantly improving the model accuracy in recognizing crossed pollen tubes. At an Intersection over Union threshold of 50%, a mean average precision of 0.949 was achieved. The performance of the model was verified using 120 testing images. The R2 value of the linear regression model using detected pollen germination frequency against the ground truth was 0.909 and that using average pollen tube length was 0.958. Further, the model was successfully applied to two other plant species, indicating a good generalizability and potential to be applied widely.
Why it matches plant phenotyping methods顕微鏡画像から花粉発芽率と花粉管長を自動推定するMask R-CNN手法を開発し、精度検証と他種への適用を行っており、植物表現型取得が研究の中心です。
abstracta Mask R-CNN model trained using microscopic images of tree peony (Paeonia suffruticosa) pollen has been proposed to rapidly detect the pollen germination rate and pollen tube length.
Reproduction assets foundThe paper's Data availability statement explicitly deposits the authors' Mask R-CNN pollen germination detection code in a public GitHub repository. Training/testing images are only available upon request (request_only), but the code asset qualifies as public and paper-specific. The makesense.ai URL is a generic third‑Code · publicThe code for local execution of the proposed model can be found in the GitHub repository ( https://github.com/Nihon-snail/Pollen_germination_detection ). Other data such as training and testing images can be provided upon reasonable request.Open asset ↗Nihon-snail/Pollen_germination_detectionlines:109-144Code / dataset availability confirmedEurope PMC · checked 7 Sept 2026
The actin cytoskeleton is essential in eukaryotes, not least in the plant kingdom where it plays key roles in cell expansion, cell division, environmental responses and pathogen defence. Yet, the precise structure-function relationships of properties of the actin network in plants are still to be unravelled, including details of how the network configuration depends upon cell type, tissue type and developmental stage. Part of the problem lies in the difficulty of extracting high-quality, quantitative measures of actin network features from microscopy data. To address this problem, we have developed DRAGoN, a novel image analysis algorithm that can automatically extract the actin network across a range of cell types, providing seventeen different quantitative measures that describe the network at a local level. Using this algorithm, we then studied a number of cases in Arabidopsis thaliana, including several different tissues, a variety of actin-affected mutants, and cells responding to powdery mildew. In many cases we found statistically-significant differences in actin network properties. In addition to these results, our algorithm is designed to be easily adaptable to other tissues, mutants and plants, and so will be a valuable asset for the study and future biological engineering of the actin cytoskeleton in globally-important crops.
Why it matches plant phenotyping methods植物画像からアクチンネットワークの定量的形質を自動抽出する画像解析アルゴリズムの開発が中心であり、植物組織・変異体・病害応答への適用も行っている。
abstractwe have developed DRAGoN, a novel image analysis algorithm that can automatically extract the actin network across a range of cell types, providing seventeen different quantitative measures that describe the network at a local level.
Reproduction assets foundThe paper's Data Availability statement explicitly deposits the complete DRAGoN analysis code (the computational tool that performs the paper's actin network extraction and phenotyping measurements) on a public GitHub repository, and the repository URL is also cited in the Results section.Code · publicData Availability: The authors confirm that all data underlying the findings are fully available without restriction. The complete code for this paper is available on a GitHub repository at https://github.com/JordanHembrow5/DRAGoN .Open asset ↗JordanHembrow5/DRAGoNlines:118-135Code / dataset availability confirmedOpenAlex · checked 14 Sept 2026
Abstract Aims Our understanding of the rhizosphere is limited by the lack of techniques for in situ live microscopy. Current techniques are either destructive or unsuitable for observing chemical changes within the pore space. To address this limitation, we have developed artificial substrates, termed smart soils, that enable the acquisition and 3D reconstruction of chemical sensors attached to soil particles. Methods The transparency of smart soils was achieved using polymer particles with refractive index matching that of water. The surface of the particles was modified both to retain water and act as a local sensor to report on pore space pH via fluorescence emissions. Multispectral signals were acquired from the particles using a light sheet microscope, and machine learning algorithms predicted the changes and spatial distribution in pH at the surface of the smart soil particles. Results The technique was able to predict pH live and in situ within ± 0.5 units of the true pH value. pH distribution could be reconstructed across a volume of several cubic centimetres around plant roots at 10 μm resolution. Using smart soils of different composition, we revealed how root exudation and pore structure create variability in chemical properties. Conclusion Smart soils captured the pH gradients forming around a growing plant root. Future developments of the technology could include the fine tuning of soil physicochemical properties, the addition of chemical sensors and improved data processing. Hence, this technology could play a critical role in advancing our understanding of complex rhizosphere processes.
Why it matches plant phenotyping methods植物根圏のpHを生体根周辺で測定・3D再構成するセンサー基盤を開発し、精度検証まで行っており、植物状態の取得方法が研究の中心である。
abstractwe have developed artificial substrates, termed smart soils, that enable the acquisition and 3D reconstruction of chemical sensors attached to soil particles.
Reproduction assets foundThe paper's Data availability statement explicitly releases the authors' software for predicting pH from light-sheet image data (the machine-learning phenotyping analysis) on the authors' public GitHub repository SENSOIL. No separate phenotype/trait dataset or image deposit is stated; supplementary material is only a 'Code · public102 Plant Soil (2024) 500:91–104
1 3
Vol:. (1234567890)
Data availability Software developped for predicting pH
from image data is available at https://github.com/LionelDu-puy/SENSOIL/tree/main/pH_Release.Declarations
Competing interest There is no competing interest.
Open Access This article is licensed under a Creative
Commons Attribution 4.0 International License, which per-
mits use, sharing, adaptation, distribution and reproduction in
any medium or format, as long as you give appropriate credit
to the original author(s) and the source,Open asset ↗SENSOILpdf-raw-page:12 lines:1-92Code / dataset availability confirmedEurope PMC · bioRxiv · checked 15 Sept 2026
MicroscopyCell / cellular structureFlowerTissue2D/3D reconstructionGrowth / development / phenology
Floral spurs are invaginations borne by perianth organs (petals and/or sepals) that have evolved repeatedly in various angiosperm clades. They typically store nectar and can limit the access of pollinators to this reward, resulting in pollination specialization that can lead to speciation in both pollinator and plant lineages. Despite the ecological and evolutionary importance of nectar spurs, the cellular mechanisms involved during spur development have only been described in detail in a handful of species, primarily with respect to epidermal cells. These studies show that the mechanisms involved are taxon-specific. Using confocal microscopy and automated 3D image analysis, we studied spur morphogenesis in Staphisagria picta (Ranunculaceae) and showed that the process is marked by an early phase of dominant cell proliferation, followed by a phase of anisotropic (directional) cell expansion. The comparison with Aquilegia , another taxon of Ranunculaceae with spurred petals, revealed that the convergence in form between the spurs of both taxa is obtained by partially similar developmental processes. The analytical pipeline designed here is an efficient method to visualize in 3D each cell of a developing organ, paving the way for future comparative studies of organ morphogenesis in multicellular eukaryotes. Highlight A new method of 3D analysis of plant tissues at the cellular level revealed that spur morphogenesis in Staphisagria picta is marked by an early phase of dominant cell proliferation, followed by a phase of anisotropic cell expansion. Floral spur development is analysed for the first time quantitatively, taking into account all tissues composing the organ, namely epidermis and parenchyma.
Why it matches plant phenotyping methods共焦点顕微鏡と自動3D画像解析による発生器官の細胞形態・増殖・異方的伸長の定量化手法が研究の中心であり、植物器官の表現型取得・解析に該当する。
abstractUsing confocal microscopy and automated 3D image analysis, we studied spur morphogenesis in Staphisagria picta (Ranunculaceae)
Reproduction assets foundThe paper's 3D segmentation/visualization pipeline (PlantSeg + MorphoLibJ + homemade Python scripts) is the paper-specific computational analysis, and the authors explicitly state the automation and visualization code is publicly available on GitHub. No separate public phenotype dataset or image deposit is stated; dataCode · public”.
235 Cell outliers, i.e. the 5% largest and smallest cells in terms of volume, were filtered out. To
236 visualize the interior of the petals, we relied on the opacity of the dots or on virtual sections.
237 The code that allowed the automation of the segmentations and the visualization of the data is
238 available on github [https://github.com/paulinedlpch/morphogenesis].Open asset ↗paulinedlpch/morphogenesispdf-layout-page:6 lines:1-57Code / dataset availability confirmedEurope PMC · checked 7 Sept 2026
Background Investigations into the growth and self-organization of plant roots is subject to fundamental and applied research in various areas such as botany, agriculture, and soil science. The growth activity of the plant tissue can be investigated by isotope labeling experiments with heavy water and subsequent detection of the deuterium in non-exchangeable positions incorporated into the plant biomass. Commonly used analytical methods to detect deuterium in plants are based on mass-spectrometry or neutron-scattering and they either suffer from elaborated sample preparation, destruction of the sample during analysis, or low spatial resolution. Confocal Raman micro-spectroscopy (CRM) can be considered a promising method to overcome the aforementioned challenges. The substitution of hydrogen with deuterium results in the measurable shift of the CH-related Raman bands. By employing correlative approaches with a high-resolution technique, such as helium ion microscopy (HIM), additional structural information can be added to CRM isotope maps and spatial resolution can be further increased. For that, it is necessary to develop a comprehensive workflow from sample preparation to data processing. Results A workflow to prepare and analyze roots of hydroponically grown and deuterium labeled Zea mays by correlative HIM-CRM micro-analysis was developed. The accuracy and linearity of deuterium detection by CRM were tested and confirmed with samples of deuterated glucose. A set of root samples taken from deuterated Zea mays in a time-series experiment was used to test the entire workflow. The deuterium content in the roots measured by CRM was close to the values obtained by isotope-ratio mass spectrometry. As expected, root tips being the most actively growing root zone had incorporated the highest amount of deuterium which increased with increasing time of labeling. Furthermore, correlative HIM-CRM analysis allowed for obtaining the spatial distribution pattern of deuterium and lignin in root cross-sections. Here, more active root zones with higher deuterium incorporation showed less lignification. Conclusions We demonstrated that CRM in combination with deuterium labeling can be an alternative and reliable tool for the analysis of plant growth. This approach together with the developed workflow has the potential to be extended to complex systems such as plant roots grown in soil.
Why it matches plant phenotyping methods植物根の成長状態を測定する相関HIM-CRMワークフローを開発し、重水素検出の精度・直線性と質量分析との一致を検証しているため、植物フェノタイピング手法が中心である。
abstractA workflow to prepare and analyze roots of hydroponically grown and deuterium labeled Zea mays by correlative HIM-CRM micro-analysis was developed.
Reproduction assets foundThe paper's data availability statement deposits the datasets generated and analyzed (CRM/HIM phenotyping measurements of deuterium-labeled maize roots) in the UFZ Data Investigation Portal, a public repository with an explicit URL. No author analysis code with a public URL is stated.Dataset · publicThe datasets generated and/or analyzed during the current study are available in the UFZ Data Investigation Portal ( https://www.ufz.de/record/dmp/archive/13952 ) repository.Open asset ↗UFZ Data Investigation Portal · archive/13952lines:198-288Code / dataset availability confirmedEurope PMC · Crossref · checked 13 Sept 2026
MicroscopyTissueCalibration / preprocessingVisualization / data management
Motivation Quantitative descriptions of multi-cellular structures from optical microscopy imaging are prime to understand the variety of three-dimensional (3D) shapes in living organisms. Experimental models of vertebrates, invertebrates and plants, such as zebrafish, killifish, Drosophila or Marchantia, mainly comprise multilayer tissues, and even if microscopes can reach the needed depth, their geometry hinders the selection and subsequent analysis of the optical volumes of interest. Computational tools to "peel" tissues by removing specific layers and reducing 3D volume into planar images, can critically improve visualization and analysis. Results We developed VolumePeeler, a versatile FIJI plugin for virtual 3D "peeling" of image stacks. The plugin implements spherical and spline surface projections. We applied VolumePeeler to perform peeling in 3D images of spherical embryos, as well as non-spherical tissue layers. The produced images improve the 3D volume visualization and enable analysis and quantification of geometrically challenging microscopy datasets. Availability ImageJ/FIJI software, source code, examples, and tutorials are openly available in https://cimt.uchile.cl/mcerda.
Why it matches plant phenotyping methods植物を含む3D組織画像の層構造を仮想的に展開し、可視化・定量化するFIJIプラグインの開発研究であり、植物組織形態の画像解析に再利用可能な手法が中心である。
abstractWe developed VolumePeeler, a versatile FIJI plugin for virtual 3D "peeling" of image stacks.
Reproduction assets foundThis is a software paper for VolumePeeler, a FIJI plugin for 3D volume peeling applied to zebrafish, killifish, and the plant model Marchantia. The authors' plugin source code and example data/tutorials are explicitly and publicly available, covering the paper's computational analysis including the Marchantia (plant) 3Code · publicSource code is available from https://github.com/busmangit/volume-peeler . Examples and video tutorials are available under Creative Commons license (CC BY-NC).Open asset ↗busmangit/volume-peelerlines:556-587Code / dataset availability confirmedOpenAlex · checked 15 Sept 2026
Field / plotMicroscopyCell / cellular structureMorphology / geometry measurement2D/3D reconstruction
Abstract The streptophyte green algal class Zygnematophyceae is the immediate sister lineage to land plants. Their special form of sexual reproduction via conjugation might have played a key role during terrestrialization. Thus, studying Zygnematophyceae and conjugation is crucial for understanding the conquest of land. Moreover, sexual reproduction features are important for species determination. We present a phylogenetic analysis of a field‐sampled Zygnema strain and analyze its conjugation process and zygospore morphology, both at the micro‐ and nanoscale, including 3D‐reconstructions of the zygospore architecture. Vegetative filament size (26.18 ± 1.07 μm) and reproductive features allowed morphological determination of Zygnema vaginatum, which was combined with molecular analyses based on rbcL sequencing. Transmission electron microscopy (TEM) depicted a thin cell wall in young zygospores, while mature cells exhibited a tripartite wall, including a massive and sculptured mesospore. During development, cytological reorganizations were visualized by focused ion beam scanning electron microscopy (FIB‐SEM). Pyrenoids were reorganized, and the gyroid cubic central thylakoid membranes, as well as the surrounding starch granules, degraded (starch granule volume: 3.58 ± 2.35 μm3 in young cells; 0.68 ± 0.74 μm3 at an intermediate stage of zygospore maturation). Additionally, lipid droplets (LDs) changed drastically in shape and abundance during zygospore maturation (LD/cell volume: 11.77% in young cells; 8.79% in intermediate cells, 19.45% in old cells). In summary, we provide the first TEM images and 3D‐reconstructions of Zygnema zygospores, giving insights into the physiological processes involved in their maturation. These observations help to understand mechanisms that facilitated the transition from water to land in Zygnematophyceae.
Why it matches plant phenotyping methodsFIB-SEM/TEMによる3D画像取得と再構成が研究の中心で、接合胞子の形態・細胞内構造・体積変化を定量的に評価しているため、画像ベースの植物表現型計測として含める。
abstractincluding 3D‐reconstructions of the zygospore architecture
Reproduction assets foundThe paper's FIB-SEM image stacks of Zygnema vaginatum zygospores (the raw imaging data underlying the 3D reconstructions) are publicly deposited as supplementary Movies S2–S4 on figshare. No author analysis code or trained models are explicitly deposited; other supporting data are available only upon request.Dataset · publicof a young Zygnema vaginatum zygospore. https://figshare.com/s/637190524e99459e5b4e .
MOVIE S3. FIB SEM generated stack of frames of an intermediate stage of Zygnema vaginatum zygospore maturation. https://figshare.com/s/30519eddaac8274263aa .
MOVIE S4. FIB SEM generated stack of frames of a mature Zygnema vaginatum zygospore. https://figshare.com/s/11bdf9f4e7ee785e7f9f .
ACKNOWLEDGMENTS
We thank Sabrina Obwegeser, MSc (University of Innsbruck, Austria) for expert technical help in TEM sectioning and image generation. We also thank Jiří Neustupa for his company and assistance during the fieldwork.
DATA AVAILABILITY STATEMENT
The supporting data of the present study are available upon requestOpen asset ↗figsharelines:180-264Code / dataset availability confirmedEurope PMC · Crossref · bioRxiv · checked 15 Sept 2026
Background Spatial mapping of transcriptional states provides valuable biological insights into cellular functions and interactions in the context of the tissue. Accurate 3D cell segmentation is a critical step in the analysis of this data towards understanding diseases and normal development in situ . Current approaches designed to automate 3D segmentation include stitching masks along one dimension, training a 3D neural network architecture from scratch, and reconstructing a 3D volume from 2D segmentations on all dimensions. However, the applicability of existing methods is hampered by inaccurate segmentations along the non-stitching dimensions, the lack of high-quality diverse 3D training data, and inhomogeneity among different dimensions; as a result, they have not been widely used in practice. Methods To address these challenges, we formulate the problem of finding cell correspondence across layers with a novel optimal transport (OT) approach. We propose CellStitch, a flexible pipeline that segments cells from 3D images without requiring large amounts of 3D training data. We further extend our method to interpolate internal slices from highly anisotropic cell images to recover isotropic cell morphology. Results We evaluated the performance of CellStitch through eight 3D plant microscopic datasets with diverse anisotropic levels and cell shapes. CellStitch substantially outperforms the state-of-the art methods on anisotropic images, and achieves comparable segmentation quality against competing methods in isotropic setting. We benchmarked and reported 3D segmentation results of all the methods with instance-level precision, recall and average precision (AP) metrics. Conclusion The proposed OT-based 3D segmentation pipeline outperformed the existing state-of-the-art methods on different datasets with nonzero anisotropy, providing high fidelity recovery of 3D cell morphology from microscopic images.
Why it matches plant phenotyping methods植物の3D細胞画像から形態を抽出するセグメンテーション手法を開発し、複数の植物データセットで性能をベンチマークしているため、植物フェノタイピング手法が中心である。
abstractWe propose CellStitch, a flexible pipeline that segments cells from 3D images
Reproduction assets foundThe paper provides explicit public availability for its CellStitch implementation and experiment-reproduction notebooks on GitHub, and evaluates on publicly available Arabidopsis thaliana image datasets, including the Arabidopsis 3D Digital Tissue Atlas hosted on OSF. These are paper-specific, public, actionable assetsCode · publicA python implementation of this method, CellStitch, is available at https://github.com/imyiningliu/300
cellstitch. The reproduction of all experiments presented herein can be accessed via https:
301
//github.com/imyiningliu/cellstitch/tree/main/notebooks.Open asset ↗imyiningliu/cellstitchpdf-raw-page:14 lines:1-70Code / dataset availability confirmedEurope PMC · checked 7 Sept 2026
Multicellular organisms result from complex developmental processes largely orchestrated through the quantitative spatiotemporal regulation of gene expression. Yet, obtaining absolute counts of messenger RNAs at a three-dimensional resolution remains challenging, especially in plants, owing to high levels of tissue autofluorescence that prevent the detection of diffraction-limited fluorescent spots. In situ hybridization methods based on amplification cycles have recently emerged, but they are laborious and often lead to quantification biases. In this article, we present a simple method based on single-molecule RNA fluorescence in situ hybridization to visualize and count the number of mRNA molecules in several intact plant tissues. In addition, with the use of fluorescent protein reporters, our method also enables simultaneous detection of mRNA and protein quantity, as well as subcellular distribution, in single cells. With this method, research in plants can now fully explore the benefits of the quantitative analysis of transcription and protein levels at cellular and subcellular resolution in plant tissues.
Why it matches plant phenotyping methods植物組織内のmRNA・タンパク質量を細胞および細胞内解像度で可視化・定量するsmFISH法の開発であり、植物の状態を測定する方法が中心的です。
abstractIn this article, we present a simple method based on single-molecule RNA fluorescence in situ hybridization to visualize and count the number of mRNA molecules in several intact plant tissues.
Reproduction assets foundThe authors openly deposited all raw microscopy images (WM-smFISH mRNA/protein imaging of Arabidopsis and barley tissues) used for their quantification pipeline on Figshare. No separate author analysis code repository with explicit availability language is stated in the supplied text.Dataset · publicAll the raw microscopy images used in this manuscript are openly available in Figshare at https://doi.org/10.6084/m9.figshare.22699132 .Open asset ↗Figshare · 10.6084/m9.figshare.22699132lines:110-216Code / dataset availability confirmedEurope PMC · checked 7 Sept 2026
Background In tissues and organisms, the coordination of neighboring cells is essential to maintain their properties and functions. Therefore, knowing which cells are adjacent is crucial to understand biological processes that involve physical interactions among them, e.g. cell migration and proliferation. In addition, some signaling pathways, such as Notch or extrinsic apoptosis, are highly dependent on cell-cell communication. While this is straightforward to obtain from membrane images, nuclei labelling is much more ubiquitous for technical reasons. However, there are no automatic and robust methods to find neighboring cells based only on nuclear markers. Results In this work, we describe Nfinder, a method to assess the cell's local neighborhood from images with nuclei labeling. To achieve this goal, we approximate the cell-cell interaction graph by the Delaunay triangulation of nuclei centroids. Then, links are filtered by automatic thresholding in cell-cell distance (pairwise interaction) and the maximum angle that a pair of cells subtends with shared neighbors (non-pairwise interaction). We systematically characterized the detection performance by applying Nfinder to publicly available datasets from Drosophila melanogaster, Tribolium castaneum, Arabidopsis thaliana and C. elegans. In each case, the result of the algorithm was compared to a cell neighbor graph generated by manually annotating the original dataset. On average, our method detected 95% of true neighbors, with only 6% of false discoveries. Remarkably, our findings indicate that taking into account non-pairwise interactions might increase the Positive Predictive Value up to + 11.5%. Conclusion Nfinder is the first robust and automatic method for estimating neighboring cells in 2D and 3D based only on nuclear markers and without any free parameters. Using this tool, we found that taking non-pairwise interactions into account improves the detection performance significantly. We believe that using our method might improve the effectiveness of other workflows to study cell-cell interactions from microscopy images. Finally, we also provide a reference implementation in Python and an easy-to-use napari plugin.
Why it matches plant phenotyping methods植物を含む組織画像から細胞近傍という形態・構造状態を自動推定する画像解析法を開発し、手動アノテーションとの比較で性能検証しているため、植物フェノタイピング手法が中心です。
abstractwe describe Nfinder, a method to assess the cell's local neighborhood from images with nuclei labeling.
Reproduction assets foundThe paper's authors provide a public reference implementation (Python code and napari plugin) of the Nfinder cell-neighborhood analysis on GitHub, and the microscopy image datasets used for validation are publicly available (CIL datasets with DOIs).Code · publicSource code of a reference implementation in Python for 2D and 3D datasets as well as a user-friendly napari plugin can be found at https://github.com/santi-rodriguez/nfinder .Open asset ↗santi-rodriguez/nfinderlines:86-111Code / dataset availability confirmedCrossref · checked 8 Sept 2026
Abstract Phytoliths constitute microscopic SiO 2 -rich biominerals formed in the cellular system of many living plants and are often preserved in soils, sediments and artefacts. Their analysis contributes significantly to the identification and study of botanical remains in (paleo)ecological and archaeological contexts. Traditional identification and classification of phytoliths rely on human experience, and as such, an emerging challenge is to automatically classify them to enhance data homogeneity among researchers worldwide and facilitate reliable comparisons. In the present study, a deep artificial neural network (NN) is implemented under the objective to detect and classify phytoliths, extracted from modern wheat ( Triticum spp.). The proposed methodology is able to recognise four phytolith morphotypes: (a) Stoma, (b) Rondel, (c) Papillate, and (d) Elongate dendritic. For the learning process, a dataset of phytolith photomicrographs was created and allocated to training, validation and testing data groups. Due to the limited size and low diversity of the dataset, an end-to-end encoder-decoder NN architecture is proposed, based on a pre-trained MobileNetV2, utilised for the encoder part and U-net, used for the segmentation stage. After the parameterisation, training and fine-tuning of the proposed architecture, it is capable to classify and localise the four classes of phytoliths in unknown images with high unbiased accuracy, exceeding 90%. The proposed methodology and corresponding dataset are quite promising for building up the capacity of phytolith classification within unfamiliar (geo)archaeological datasets, demonstrating remarkable potential towards automatic phytolith analysis.
Why it matches plant phenotyping methods植物由来の植物珪酸体を画像から検出・分類する深層学習手法とデータセットの開発が研究の中心であり、植物形態情報の取得・抽出に該当する。
abstracta deep artificial neural network (NN) is implemented under the objective to detect and classify phytoliths, extracted from modern wheat
Reproduction assets foundThe paper's phytolith photomicrograph dataset (annotated images of four morphotypes from modern wheat) is explicitly stated to be publicly available on Kaggle. Code and trained NNs are mentioned as contributions but no public repository URL is provided, so only the dataset qualifies.Dataset · publicData availability
The dataset of the current study is publicly available in the Kaggle platform: https://www.kaggle.com/datasets/georgepetrakis/phytolith-photomicrographs .Open asset ↗Kaggle · georgepetrakis/phytolith-photomicrographslines:130-152Code / dataset availability confirmedCrossref · checked 14 Sept 2026
Viruses are key players in marine ecosystems where they infect abundant marine microbes. RNA viruses are emerging as key members of the marine virosphere. They have recently been identified as a potential source of mortality in diatoms, a group of microalgae that accounts for roughly 40% of the primary production in the ocean. Despite their likely importance, their impacts on host populations and ecosystems remain difficult to assess. In this study, we introduce an innovative approach that combines automated 3D confocal microscopy with quantitative image analysis and physiological measurements to expand our understanding of viral infection. We followed different stages of infection of the bloom-forming diatom Guinardia delicatula by the RNA virus GdelRNAV-04 until the complete lysis of the host. From 20h after infection, we observed quantifiable changes in subcellular host morphology and biomass. Our microscopy monitoring also showed that viral infection of G. delicatula induced the formation of auxospores as a probable defense strategy against viruses. Our method enables the detection of discriminative morphological features on the subcellular scale and at high throughput for comparing populations, making it a promising approach for the quantification of viral infections in the field in the future.
Why it matches plant phenotyping methods自動3D共焦点顕微鏡と定量画像解析を組み合わせ、感染に伴う珪藻の細胞形態・バイオマスを高スループットに定量する手法が研究の中心である。
abstractwe introduce an innovative approach that combines automated 3D confocal microscopy with quantitative image analysis and physiological measurements
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Code · publicThe full annotated script can be found on https://github.com/mariescopy/Guinardia_ViralInfectionOpen asset ↗mariescopy/Guinardia_ViralInfectionlines:286-318Code / dataset availability confirmedEurope PMC · checked 7 Sept 2026
Background Live imaging is the gold standard for determining how cells give rise to organs. However, tracking many cells across whole organs over large developmental time windows is extremely challenging. In this work, we provide a comparably simple method for confocal live imaging entire Arabidopsis thaliana first leaves across early development. Our imaging method works for both wild-type leaves and the complex curved leaves of the jaw-1D mutant. Results We find that dissecting the cotyledons, affixing a coverslip above the samples and mounting samples with perfluorodecalin yields optimal imaging series for robust cellular and organ level analysis. We provide details of our complementary image processing steps in MorphoGraphX software for segmenting, tracking lineages, and measuring a suite of cellular properties. We also provide MorphoGraphX image processing scripts we developed to automate analysis of segmented images and data presentation. Conclusions Our imaging techniques and processing steps combine into a robust imaging pipeline. With this pipeline we are able to examine important nuances in the cellular growth and differentiation of jaw-D versus WT leaves that have not been demonstrated before. Our pipeline is approachable and easy to use for leaf development live imaging.
Why it matches plant phenotyping methodsArabidopsis葉の生細胞イメージング、画像処理、細胞追跡・形質測定を統合した再利用可能な表現型解析パイプラインの開発が中心である。
abstractIn this work, we provide a comparably simple method for confocal live imaging entire Arabidopsis thaliana first leaves across early development.
Reproduction assets foundThe paper publicly deposits its live-imaging datasets (confocal imaging data for the figures) on OSF under CC-BY 4.0, and its MorphoGraphX/R analysis scripts on the authors' GitHub repositories, all explicitly linked in the Availability of data and materials section.Dataset · publicData for Figs. 1 , 2 , 3 , 4 , 5 A, B is available at https://doi.org/10.17605/OSF.IO/V2TKWOpen asset ↗OSF · 10.17605/OSF.IO/V2TKWlines:139-172Dataset · publicData for Figs. 5 C, 6 and 7 is available at https://doi.org/10.17605/OSF.IO/D7X3YOpen asset ↗OSF · 10.17605/OSF.IO/D7X3Ylines:139-172Code · publicavailable at https://github.com/kateharline/live_img_paper , https://github.com/kateharline/roeder_lab_projects/tree/master/imagej_scripts and https://github.com/kateharline/jawd-paperOpen asset ↗github.com/kateharline/roeder_lab_projectslines:139-172Code · publicavailable at https://github.com/kateharline/live_img_paper , https://github.com/kateharline/roeder_lab_projects/tree/master/imagej_scripts and https://github.com/kateharline/jawd-paperOpen asset ↗github.com/kateharline/jawd-paperlines:139-172Code / dataset availability confirmedEurope PMC · bioRxiv · checked 15 Sept 2026
A bstract The actin cytoskeleton is essential in eukaryotes, not least in the plant kingdom where it plays key roles in cell expansion, cell division, environmental responses and pathogen defence. Yet, the precise structure-function relationships of properties of the actin network in plants are still to be unravelled, including details of how the network configuration depends upon cell type, tissue type and developmental stage. Part of the problem lies in the difficulty of extracting high-quality, three-dimensional, quantitative measures of actin network features from microscopy data. To address this problem, we have developed DRAGoN, a novel image analysis algorithm that can automatically extract the actin network across a range of cell types, providing seventeen different quantitative measures that describe the network at a local level. Using this algorithm, we then studied a number of cases in Arabidopsis thaliana , including several different tissues, a variety of actin-affected mutants, and cells responding to powdery mildew. In many cases we found statistically-significant differences in actin network properties. In addition to these results, our algorithm is designed to be easily adaptable to other tissues, mutants and plants, and so will be a valuable asset for the study and future biological engineering of the actin cytoskeleton in globally-important crops.
Why it matches plant phenotyping methods植物の顕微鏡画像からアクチンネットワークの構造特性を自動抽出する画像解析手法を開発しており、植物状態の定量的表現型取得が中心である。
abstractwe have developed DRAGoN, a novel image analysis algorithm that can automatically extract the actin network across a range of cell types, providing seventeen different quantitative measures that describe the network at a local level.
Reproduction assets foundThe paper's DRAGoN actin-network extraction algorithm (authors' analysis code) is explicitly stated to be freely available and open source on GitHub. No public phenotype dataset or image deposit is described in the supplied blocks.Code · publicery small amount. A much larger data set or perhaps an
artificial stimulation of the immune response (e.g. a microneedle assay[80]) may help in discerning these changes in
more detail.
To facilitate further development or optimisation for particular data sets, we have made the DRAGoN software freely
available and open source at https://github.com/JordanHembrow5/DRAGoN. The flexibility and non-specificity of this
tool is one of its main advantages and should enable it to be useful in a range of organisms, mutants, tissues, cell types
and environments. A number of key parameters (particularly those for the filtering and skeletonisation steps) can be
adjusted to best fit a given image modalityOpen asset ↗JordanHembrow5/DRAGoNpdf-layout-page:16 lines:1-48Code / dataset availability confirmedEurope PMC · checked 8 Sept 2026
Plant disease classification is quite complex and, in most cases, requires trained plant pathologists and sophisticated labs to accurately determine the cause. Our group for the first time used microscopic images (×30) of tomato plant diseases, for which representative plant samples were diagnostically validated to classify disease symptoms using non-coding deep learning platforms (NCDL). The mean F1 scores (SD) of the NCDL platforms were 98.5 (1.6) for Amazon Rekognition Custom Label, 93.9 (2.5) for Clarifai, 91.6 (3.9) for Teachable Machine, 95.0 (1.9) for Google AutoML Vision, and 97.5 (2.7) for Microsoft Azure Custom Vision. The accuracy of the NCDL platform for Amazon Rekognition Custom Label was 99.8% (0.2), for Clarifai 98.7% (0.5), for Teachable Machine 98.3% (0.4), for Google AutoML Vision 98.9% (0.6), and for Apple CreateML 87.3 (4.3). Upon external validation, the model's accuracy of the tested NCDL platforms dropped no more than 7%. The potential future use for these models includes the development of mobile- and web-based applications for the classification of plant diseases and integration with a disease management advisory system. The NCDL models also have the potential to improve the early triage of symptomatic plant samples into classes that may save time in diagnostic lab sample processing.
Why it matches plant phenotyping methodsトマト葉の顕微鏡画像から病徴を分類する深層学習モデルを開発・比較し、外部検証まで実施しており、植物病害状態の表現型取得が中心である。
abstractUpon external validation, the model's accuracy of the tested NCDL platforms dropped no more than 7%.
Reproduction assets foundThe paper's data availability statement explicitly deposits the microscopic tomato disease image dataset used for training the NCDL models in a public GitHub repository, which is a paper-specific, publicly actionable asset.Dataset · publicThe datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found below: https://github.com/manoj044/Tomato_microscopic_images.git .Open asset ↗https://github.com/manoj044/Tomato_microscopic_images.gitlines:993-1025Code / dataset availability confirmedEurope PMC · Crossref · checked 13 Sept 2026
Background and aims The surface area of mesophyll cells (Smes) and chloroplasts (Sc) facing the intercellular airspace (IAS) are important parameters for estimating photosynthetic activity from leaf anatomy. Although Smes and Sc are estimated based on the shape assumption of mesophyll cells (MCs), it is questionable if the assumption is correct for rice MCs with concave-convex surfaces. Therefore, in this study, we establish a reconstruction method for the 3-D representation of the IAS in rice leaf tissue to calculate the actual Smes and Sc with 3-D images and to determine the correct shape assumption for the estimation of Smes and Sc based on 2-D section images. Methods We used serial section light microscopy to reconstruct 3-D representations of the IAS, MCs and chloroplasts in rice leaf tissue. Actual Smes and Sc values obtained from the 3-D representation were compared with those estimated from the 2-D images to find the correct shape-specific assumption (oblate or prolate spheroid) in different orientations (longitudinal and transverse sections) using the same leaf sample. Key results The 3-D representation method revealed that volumes of the IAS and MCs accounted for 30 and 70 % of rice leaf tissue excluding epidermis, respectively, and the volume of chloroplasts accounted for 44 % of MCs. The shape-specific assumption on the sectioning orientation affected the estimation of Smes and Sc using 2-D section images with discrepancies of 10-38 %. Conclusions The 3-D representation of rice leaf tissue was successfully reconstructed using serial section light microscopy and suggested that estimation of Smes and Sc of the rice leaf is more accurate using longitudinal sections with MCs assumed as oblate spheroids than using transverse sections with MCs as prolate spheroids.
Why it matches plant phenotyping methodsイネ葉組織の3D再構成法を開発し、2D画像による葉肉細胞・葉緑体の表面積推定と比較検証しており、植物形質の取得・推定法が中心である。
abstractwe establish a reconstruction method for the 3-D representation of the IAS in rice leaf tissue to calculate the actual Smes and Sc with 3-D images
Reproduction assets foundThe paper's supplementary materials include 3-D reconstruction videos of the IAS and chloroplast regions facing the IAS (Videos S1 and S2) and a table of shape assumptions used for curvature correction factors (Table S1), all directly reproducing this paper's phenotyping measurements. These are hosted online via the OyDataset · publicSupplementary data are available online at https://academic.oup.com/aob and consist of the following:Open asset ↗lines:139-146Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
The leaf epidermis is the interface between a plant and its environment. The epidermis is highly variable in morphology, with links to both phylogeny and environment, and this diversity is relevant to several fields, including physiology, functional traits, palaeobotany, taxonomy and developmental biology. Describing and measuring leaf epidermal traits remains challenging. Current approaches are either extremely labour-intensive and not feasible for large studies or limited to measurements of individual cells. Here, we present a method to characterise individual cell size, shape (including the effect of neighbouring cells) and arrangement from light microscope images. We provide the first automated characterisation of cell arrangement (from traced images) as well as multiple new shape characteristics. We have implemented this method in an R package, epidermalmorph, and provide an example workflow using this package, which includes functions to evaluate trait reliability and optimal sampling effort for any given group of plants. We demonstrate that our new metrics of cell shape are independent of gross cell shape, unlike existing metrics. epidermalmorph provides a broadly applicable method for quantifying epidermal traits that we hope can be used to disentangle the fundamental relationships between form and function in the leaf epidermis.
Why it matches plant phenotyping methods葉の表皮細胞の形状・配置などの植物形質を顕微鏡画像から自動定量する手法とRパッケージを開発しており、方法が研究の中心である。
abstractHere, we present a method to characterise individual cell size, shape (including the effect of neighbouring cells) and arrangement from light microscope images.
Reproduction assets foundThe paper's authors publicly released their analysis code, the epidermalmorph R package, on GitHub with documentation and tutorials, including example data ('podocarps') in the package. The expanded phenotype dataset is not yet available (request_only), but the code asset qualifies as public and paper-specific.Code · publicThe R package (including installation instructions) is on GitHub ( https://github.com/matildabrown/epidermalmorph ) with accompanying documentation and tutorials ( https://matildabrown.github.io/epidermalmorph/ ). Example data are available in the R package as the dataset ‘ podocarps ’. Figs 7 and 8 are examples of the output produced by this package; the expanded dataset used to generate these figures forms part of a forthcoming study (exOpen asset ↗matildabrown/epidermalmorphlines:188-571Code · publicThe R package (including installation instructions) is on GitHub ( https://github.com/matildabrown/epidermalmorph ) with accompanying documentation and tutorials ( https://matildabrown.github.io/epidermalmorph/ ). Example data are available in the R package as the dataset ‘ podocarps ’. Figs 7 and 8 are examples of the output produced by this package; the expanded dataset used to generate these figures forms part of a forthcoming study (expected publication in early 2023) and will be made available with this future paper. Please contaOpen asset ↗lines:188-571Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
Background Researchers interested in the seed size of rapeseed need to quantify the cell size and number of cells in the seed coat, embryo and silique wall. Scanning electron microscope-based methods have been demonstrated to be feasible but laborious and costly. After image preparation, the cell parameters are generally evaluated manually, which is time consuming and a major bottleneck for large-scale analysis. Recently, two machine learning-based algorithms, Trainable Weka Segmentation (TWS) and Cellpose, were released to overcome this long-standing problem. Moreover, the MorphoLibJ and LabelsToROIs plugins in Fiji provide user-friendly tools to deal with cell segmentation files. We attempted to verify the practicability and efficiency of these advanced tools for various types of cells in rapeseed. Results We simplified the current image preparation procedure by skipping the fixation step and demonstrated the feasibility of the simplified procedure. We developed three methods to automatically process multicellular images of various tissues in rapeseed. The TWS-Fiji (TF) method combines cell detection with TWS and cell measurement with Fiji, enabling the accurate quantification of seed coat cells. The Cellpose-Fiji (CF) method, based on cell segmentation with Cellpose and quantification with Fiji, achieves good performance but exhibits systematic error. By removing border labels with MorphoLibJ and detecting regions of interest (ROIs) with LabelsToROIs, the Cellpose-MorphoLibJ-LabelsToROIs (CML) method achieves human-level performance on bright-field images of seed coat cells. Intriguingly, the CML method needs very little manual calibration, a property that makes it suitable for massive-scale image processing. Through a large-scale quantitative evaluation of seed coat cells, we demonstrated the robustness and high efficiency of the CML method at both the single-cell level and the sample level. Furthermore, we extended the application of the CML method to developing seed coat, embryo and silique wall cells and acquired highly precise and reliable results, indicating the versatility of this method for use in multiple scenarios. Conclusions The CML method is highly accurate and free of the need for manual correction. Hence, it can be applied for the low-cost, high-throughput quantification of diverse cell types in rapeseed with high efficiency. We envision that this method will facilitate the functional genomics and microphenomics studies of rapeseed and other crops.
Why it matches plant phenotyping methodsアブラナの種皮・胚・長角果壁の細胞形質を画像から自動定量する手法を開発・検証しており、フェノタイピング手法が研究の中心です。
abstractWe developed three methods to automatically process multicellular images of various tissues in rapeseed.
Live imaging is the gold standard for determining how cellular development gives rise to organs. However, tracking all individual cells across whole organs over large developmental time windows is extremely challenging. In this work, we provide a comparably simple method for confocal live imaging of Arabidopsis thaliana first leaves across early development. Our imaging method works for both wild-type leaves and the complex curved leaves of the jaw-1D mutant. We find that dissecting the cotyledons, affixing a coverslip above the samples and mounting samples with perfluorodecalin yields optimal imaging series for robust cellular and organ level analysis. We provide details of our complementary image processing steps in MorphGraphX software for segmenting cells, tracking the cell lineages, and measuring a suite of cellular growth properties. We also provide MorphoGraphX image processing scripts that we developed to automate analysis of segmented images and data presentation. Our imaging techniques and processing steps combine into a robust imaging pipeline. With this pipeline we are able to examine important nuances in the cellular growth and differentiation of jaw-D versus WT leaves that have not been demonstrated before. Our pipeline is a practical starting place for researchers new to live imaging plant leaves, but also to anyone interested in improving the throughput and reliability of their live imaging process.
Why it matches plant phenotyping methods葉全体の共焦点ライブイメージング、細胞セグメンテーション・系譜追跡・成長特性測定を統合した実用的な表現型解析パイプラインの開発であり、方法が研究の中心です。
abstractIn this work, we provide a comparably simple method for confocal live imaging of Arabidopsis thaliana first leaves across early development.
Reproduction assets foundThe paper's phenotyping analysis code is publicly available in authors' GitHub repositories: MorphoGraphX processing/quantification scripts (iterative_growth_and_measures.py, multi_resize.py, batch_tiff.py) in roeder_lab_projects/mgx_scripts, ImageJ scripts, and R analysis/figure scripts in live_img_paper and jawdPaperCode · publice heat map representations of the
data with standardized parameters across time point comparisons and replicates (Video 4).
Data analysis
All data processing, analysis and plotting was performed in RStudio (2020; 2021). Scripts used
to process the data and create figures are enclosed as Supplemental Information and available at
https://github.com/kateharline/live_img_paper, https://github.com/kateharline/roeder_lab_proj-ects/tree/master/imagej_scripts and https://github.com/kateharline/jawd-paper.Data availability
Imaging data will be deposited XXXX.
Funding
Kate Harline was supported by NSF Graduate Research Fellowship (DGE-1650441). This work
was funded by NSF MCB-2203275 (AHKR), The SchwaOpen asset ↗kateharline/live_img_paperpdf-raw-page:20 lines:1-63Code · publicndardized parameters across time point comparisons and replicates (Video 4).
Data analysis
All data processing, analysis and plotting was performed in RStudio (2020; 2021). Scripts used
to process the data and create figures are enclosed as Supplemental Information and available at
https://github.com/kateharline/live_img_paper, https://github.com/kateharline/roeder_lab_proj-ects/tree/master/imagej_scripts and https://github.com/kateharline/jawd-paper.Data availability
Imaging data will be deposited XXXX.
Funding
Kate Harline was supported by NSF Graduate Research Fellowship (DGE-1650441). This work
was funded by NSF MCB-2203275 (AHKR), The Schwartz Research Fund Award (AHKR), and
the NationaOpen asset ↗kateharline/roeder_lab_proj-ectspdf-raw-page:20 lines:1-63Code · publicnalysis
All data processing, analysis and plotting was performed in RStudio (2020; 2021). Scripts used
to process the data and create figures are enclosed as Supplemental Information and available at
https://github.com/kateharline/live_img_paper, https://github.com/kateharline/roeder_lab_proj-ects/tree/master/imagej_scripts and https://github.com/kateharline/jawd-paper.Data availability
Imaging data will be deposited XXXX.
Funding
Kate Harline was supported by NSF Graduate Research Fellowship (DGE-1650441). This work
was funded by NSF MCB-2203275 (AHKR), The Schwartz Research Fund Award (AHKR), and
the National Institute Of General Medical Sciences of the National Institutes of Health under
Open asset ↗kateharline/jawd-paperpdf-raw-page:20 lines:1-63Code / dataset availability confirmedEurope PMC · checked 15 Sept 2026
Field / plotMicroscopyClassificationCountingMorphology / geometry measurement
Stingless bees are effective pollinators of native tropical flora. Their environmental service maintains flow of pollen through pollination, increase reproductive success and influence genetic structure in plants. The management of stingless bees "meliponiculture", is an activity limited to the countryside in Ecuador. The lack of knowledge of their managers about pollen resources can affect the correct maintenance/production of nests. The objective is to identify botanical families and genera of pollen grains collected by stingless bees by morphological features and differentiate potential species using geometric morphometry. Thirty-six pot pollen samples were collected from three Ecuadorian provinces located in two climatically different zones. Pollen type identification was based on the Number, Position, Character system. Using morphological features, the families and genera were established. Morphometry landmarks were used to show variation for species differentiation. Abundance, diversity, similarity and dominance indices were established by counting pollen grains, as well as spatial distribution relationships by means of Poisson regression. Forty-six pollen types were determined in two study areas, classified into 27 families and 18 genera. In addition, it was possible to identify more than one species, classified within the same family and genus, thanks to morphometric analysis. 1148 ± 799 (max 4211; min 29) pollen grains were counting in average. The diversity showed a high richness, low dominance and similarity between pollen resources. Families Melastomataceae and Asteraceae, genera Miconia and Bidens, were found as the main pollen resources. The stingless bee of this study are mostly generalist as shown the interaction network. The results of the present survey showed that stingless bees do not collect pollen from a single species, although there is evidence of a predilection for certain plant families. The diversity indexes showed high richness but low uniformity in the abundance of each family identified. The results of the study are also meaningful to the meliponiculture sector as there is a need to improve management practices to preserve the biodiversity and the environment.
Why it matches plant phenotyping methods花粉の形態特徴と幾何学的形態計測を用いて植物種・分類群を識別する方法が研究の中心であり、植物器官の形態形質を抽出している。
abstractMorphometry landmarks were used to show variation for species differentiation.
Reproduction assets foundThe paper's pollen phenotyping measurements (morphological descriptions, size/shape parameters, and diversity/dominance/similarity indices) are published as public supporting information files on PLOS ONE. No author analysis code or trained models are deposited; the S1 Table is a literature review compilation rather a Supplement · publicmilies features used as pollen and nectar resources by stingless bees [ 41 – 91 , 126 ].
https://doi.org/10.1371/journal.pone.0272580.s001
(DOCX)
S2 Table. Morphological description, and their parameters, of pollen grains with the highest representation in the three study areas [ 43 , 47 , 52 , 84 , 86 , 99 , 100 , 118 – 129 ].
https://doi.org/10.1371/journal.pone.0272580.s002
(DOCX)
S3 Table. Indexes explaining chart.
Comparison between the diversity, dominance and similarity indices in each study area.
https://doi.org/10.1371/journal.pone.0272580.s003
(DOCX)
Acknowledgments
This work would not have been accomplished without the help of Ecuadorian meliponicultors, regular and thesis studentOpen asset ↗10.1371/journal.pone.0272580.s002lines:320-367Code / dataset availability confirmedEurope PMC · OpenAlex · Crossref · checked 8 Sept 2026
In the absence of pollination, female reproductive organs senesce, leading to an irrevocable loss in the reproductive potential of the flower, which directly affects seed set. In self-pollinating crops like wheat (Triticum aestivum), the post-anthesis viability of unpollinated carpels has been overlooked, despite its importance for hybrid seed production systems. To advance our knowledge of carpel development in the absence of pollination, we created a high-throughput phenotyping approach to quantify stigma and ovary morphology. We demonstrate the suitability of the approach, which uses light-microscopy imaging and machine learning, for the analysis of floral organ traits in field-grown plants using fresh and fixed samples. We show that the unpollinated carpel undergoes a well-defined initial growth phase, followed by a peak phase in which stigma area reaches its maximum and the radial expansion of the ovary slows, and a final deterioration phase. These developmental dynamics were consistent across years and could be used to classify male-sterile cultivars. This phenotyping approach provides a new tool for examining carpel development, which we hope will advance research into female fertility of wheat.
Why it matches plant phenotyping methodsコムギの柱頭・子房形態を定量化する高スループット画像・機械学習手法の開発と適用が研究の中心であり、植物表現型取得法として明確に該当する。
abstractwe created a high-throughput phenotyping approach to quantify stigma and ovary morphology
Reproduction assets foundThe paper's authors publicly deposited both the analysis code (CNN training/implementation scripts and R scripts) on GitHub and the carpel image/training/validation datasets on Earlham OpenData, directly reproducing this paper's wheat carpel phenotyping measurements and analysis.Code · publicTraining codes used for the development of the CNNs, adapted stigma and ovary CNNs, and R scripts used for data curation and visualisation can be found at https://github.com/Uauy-Lab/ML-carpel_traitsOpen asset ↗Uauy-Lab/ML-carpel_traitslines:122-188Dataset · publicDatasets for the training and validation of the models and raw images used for the different experimental analyses are freely available at https://opendata.earlham.ac.uk/wheat/under_license/toronto/Millan-Blanquez_etal_2022_machine-learning-carpel-traits/Open asset ↗lines:122-188Code / dataset availability confirmedEurope PMC · checked 8 Sept 2026
Mitochondria form highly dynamic populations in the cells of plants (and almost all eukaryotes). The characteristics and benefits of this collective behaviour, and how it is influenced by nuclear features, remain to be fully elucidated. Here, we use a recently developed quantitative approach to reveal and analyse the physical and collective 'social' dynamics of mitochondria in an Arabidopsis msh1 mutant where the organelle DNA maintenance machinery is compromised. We use a newly created line combining the msh1 mutant with mitochondrially targeted green fluorescent protein (GFP), and characterize mitochondrial dynamics with a combination of single-cell time-lapse microscopy, computational tracking, and network analysis. The collective physical behaviour of msh1 mitochondria is altered from that of the wild type in several ways: mitochondria become less evenly spread, and networks of inter-mitochondrial encounters become more connected, with greater potential efficiency for inter-organelle exchange-reflecting a potential compensatory mechanism for the genetic challenge to the mitochondrial DNA population, supporting more inter-organelle exchange. We find that these changes are similar to those observed in friendly, where mitochondrial dynamics are altered by a physical perturbation, suggesting that this shift to higher connectivity may reflect a general response to mitochondrial challenges, where physical dynamics of mitochondria may be altered to control the genetic structure of the mtDNA population.
Why it matches plant phenotyping methods植物ミトコンドリアの動態を、タイムラプス顕微鏡・計算追跡・ネットワーク解析で定量化する手法の実質的な適用であり、単なる生物学的ルーチン測定ではない。
abstractcharacterize mitochondrial dynamics with a combination of single-cell time-lapse microscopy, computational tracking, and network analysis.
Reproduction assets foundThe paper states that all data and analysis code for the mitochondrial dynamics phenotyping are publicly available on the authors' GitHub repository, which matches an allowed URL.Code · publicAll data and analysis codes are available from Github at https://github.com/StochasticBiology/plant-mito-dynamicsOpen asset ↗StochasticBiology/plant-mito-dynamicslines:109-163Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
WheatField / plotMicroscopyRootMorphology / geometry measurementRoot system architecture
Root hairs play an important role in absorbing water and nutrients in crop plants. Here we optimized high-throughput root hair length (RHL) and root hair density (RHD) phenotyping in wheat using a portable Dinolite™ microscope. A collection of 24 century wide spring wheat cultivars released between 1911 and 2016 were phenotyped for RHL and RHD. The results revealed significant variations for both traits with five and six-fold variation for RHL and RHD, respectively. RHL ranged from 1.01 mm to 1.77 mm with an average of 1.39 mm, and RHD ranged from 17.08 mm -2 to 20.8 mm -2 with an average of 19.6 mm -2 . Agronomic and physiological traits collected from five different environments and their best linear unbiased predictions (BLUPs) were correlated with RHL and RHD, and results revealed that relative-water contents (RWC), biomass and grain per spike (GpS) were positively correlated with RHL in both water-limited and well-watered conditions. While RHD was negatively correlated with grain yield (GY) in four environments and their BLUPs. Both RHL and RHD had positive correlation indicating the possibility of simultaneous selection of both phenotypes during wheat breeding. The expression pattern of TaRSL4 gene involved in regulation of root hair length was determined in all 24 wheat cultivars based on RNA-seq data, which indicated the differentially higher expression of the A- and D- homeologues of the gene in roots, while B-homeologue was consistently expressed in both leaf and roots. The results were validated by qRT-PCR and the expression of TaRSL4 was consistently high in rainfed cultivars such as Chakwal-50, Rawal-87, and Margallah-99. Overall, the new phenotyping method for RHL and RHD along with correlations with morphological and physiological traits in spring wheat cultivars improved our understanding for selection of these phenotypes in wheat breeding.
Why it matches plant phenotyping methods携帯型顕微鏡を用いたコムギ根毛長・密度のハイスループット表現型測定法を最適化し、品種で実証しているため、根形態フェノタイピング手法が中心である。
abstractHere we optimized high-throughput root hair length (RHL) and root hair density (RHD) phenotyping in wheat using a portable Dinolite™ microscope.
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Supplement · publicTable S1: Name of the cultivars, pedigree, year of release and raw phenotypic data used in this study.Open asset ↗lines:71-196Code / dataset availability confirmedEurope PMC · bioRxiv · checked 15 Sept 2026
Background Cyst nematodes are one of the major groups of plant-parasitic nematode, responsible for considerable crop losses worldwide. Improving genetic resources, and therefore resistant cultivars, is an ongoing focus of many pest management strategies. One of the major bottlenecks in identifying the plant genes that impact the infection, and thus the yield, is phenotyping. The current available screening method is slow, has unidimensional quantification of infection limiting the range of scorable parameters, and does not account for phenotypic variation of the host. The ever-evolving field of computer vision may be the solution for both the above-mentioned issues. To utilise these tools, a specialised imaging platform is required to take consistent images of nematode infection in quick succession. Results Here, we describe an open-source, easy to adopt, imaging hardware and trait analysis software method based on a pre-existing nematode infection screening method in axenic culture. A cost-effective, easy-to-build and -use, 3D-printed imaging device was developed to acquire images of the root system of Arabidopsis thaliana infected with the cyst nematode Heterodera schachtii , replacing costly microscopy equipment. Coupling the output of this device to simple analysis scripts allowed the measurement of some key traits such as nematode number and size from collected images, in a semi-automated manner. Additionally, we used this combined solution to quantify an additional trait, root area before infection, and showed both the confounding relationship of this trait on nematode infection and a method to account for it. Conclusion Taken together, this manuscript provides a low-cost and open-source method for nematode phenotyping that includes the biologically relevant nematode size as a scorable parameter, and a method to account for phenotypic variation of the host. Together these tools highlight great potential in aiding our understanding of nematode parasitism.
Why it matches plant phenotyping methods植物寄生性線虫感染の画像取得・解析を自動化する低コストの装置とソフトウェアを開発し、線虫数・サイズおよび根面積を測定する手法が中心であるため。
abstractHere, we describe an open-source, easy to adopt, imaging hardware and trait analysis software method
Reproduction assets foundThe paper's ImageJ analysis scripts (root surface area, colored-agar variant, leaf surface count) and a custom Python color-normalization script are explicitly deposited in the authors' public GitHub repository (OlafKranse/A_low_cost_imaging_tower), directly reproducing this paper's phenotyping analysis. No phenotype/тCode · publici.org/10.1101/2022.07.14.500020; this version posted July 15, 2022. The copyright holder for this preprint
(which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is
made available under a CC-BY 4.0 International license.
described in the script
(https://github.com/OlafKranse/A_low_cost_imaging_tower/blob/main/Imaging and
Analyses/automated_root_surface_area.ijm). A slightly adjusted script was used for plates
containing dye (https://github.com/OlafKranse/A_low_cost_imaging_tower/blob/main/Imaging and
Analyses/automated_root_surface_area_colored_agar.ijm). The root surface area for all the images
in the folderOpen asset ↗OlafKranse/A_low_cost_imaging_towerpdf-layout-page:6 lines:1-37Code · publicing and quantifiable traits
Automatic counting was performed on images taken as described above. Depending on the
treatment a different script was used to calculate the number and size of females. Before isolation,
the colour histogram for all images was normalised to the first image in the dataset using a custom
python script (https://github.com/OlafKranse/A_low_cost_imaging_tower/tree/main/Imaging and
Analyses/Normalise colour). The images were then processed in ImageJ for two different nematode
life stages: i) tanned cyst nematodes; ii) female nematodes.Open asset ↗OlafKranse/A_low_cost_imaging_towerpdf-layout-page:6 lines:1-37Code / dataset availability confirmedEurope PMC · Crossref · checked 8 Sept 2026
Abstract Accurate delineation of plant cell organelles from electron microscope images is essential to understand subcellular behaviors and functions. Here, we develop a deep learning pipeline, organelle segmentation network (OrgSegNet) for pixel-wise segmentation to identify chloroplasts, mitochondria, nuclei, and vacuoles. OrgSegNet was evaluated on a large manually-annotated dataset of 6371 organelles collected from 13 plant species, and achieved a state-of-the-art segmentation performance of these organelles. To generalize the morphological characteristics of plant organelles, we defined three morphological metrics (shape-complexity, electron-density, and area), and released an open-source web tool “Plantorgan Hunter” allowing quantitative profiling of subcellular morphology. The functionalities of Plantorgan Hunter can be easily operated, and we believe that it will increase the efficiency and productivity of plant subcellular morphological characteristics for the plant science community.
Why it matches plant phenotyping methods植物細胞小器官の画像セグメンテーションと形態指標抽出を開発・評価し、データセットと公開ツールまで提供する、明確な植物フェノタイピング手法研究。
abstractHere, we develop a deep learning pipeline, organelle segmentation network (OrgSegNet) for pixel-wise segmentation to identify chloroplasts, mitochondria, nuclei, and vacuoles.
Reproduction assets foundThe paper publicly releases its manually-annotated TEM organelle dataset (Science Data Bank), the OrgSegNet code and trained models (GitHub), and a web tool (cropopen.com) for quantitative subcellular morphology profiling.Dataset · publicThe plant organelle dataset for the current study is available in the Sicence Data
Bank repository, https://www.scidb.cn/s/EBvqei.Open asset ↗pdf-page:27 lines:1-31Code / dataset availability confirmedEurope PMC · checked 15 Sept 2026
MicroscopyTissueMorphology / geometry measurementGrowth / development / phenology
In this study, we present a detailed protocol for live imaging and quantitative analysis of floral meristem development in Aquilegia coerulea , a member of the buttercup family (Ranunculaceae). Using confocal microscopy and the image analysis software MorphoGraphX, we were able to examine the cellular growth dynamics during floral organ primordia initiation, and the transition from floral meristem proliferation to termination. This protocol provides a powerful tool to study the development of the meristem and floral organ primordia, and should be easily adaptable to many plant lineages, including other emerging model systems. It will allow researchers to explore questions outside the scope of common model systems.
Why it matches plant phenotyping methods植物の花序メリステムを対象に、共焦点ライブイメージングと画像解析による細胞成長動態・器官原基形成の定量プロトコルを提示しており、表現型取得法が中心である。
abstractwe present a detailed protocol for live imaging and quantitative analysis of floral meristem development in Aquilegia coerulea
Reproduction assets foundThe protocol shares two original .czi confocal image files from the authors' own Aquilegia floral meristem study via a public Google Drive link, used to reproduce the paper's MorphoGraphX phenotyping analysis. Generic software links (Fiji/ImageJ, MorphoGraphX) are excluded as non-paper-specific.Dataset · publice stored, extracted, and processed. Here, we focus on the steps and parameters that are specific to processing confocal images of Aquilegia floral meristems, and steps to reproduce figures in Min et al. (2022). We will use two original .czi files from our study as an example, which can be downloaded from this google drive link:
https://drive.google.com/drive/folders/1WjaCieLGrnTW7d51143b8HOn-dYmsMU-?usp=sharing
Images of individual time points will be processed separately first, then loaded together for lineage tracing (details in the following section Parent Labeling & Lineage Tracing).
Software installation and equipment setup
Download the newest version of MGX from
https://morphographx.orOpen asset ↗lines:202-231Code / dataset availability confirmedbioRxiv · checked 8 Sept 2026
X-ray fluorescence spectroscopy (XRF) is a powerful technique for the in vivo assessment of plant tissues. However, the potential X-ray exposure damages might affect the structure and elemental composition of living plant tissues leading to artefacts in the recorded data. Herein, we exposed soybean (Glycine max (L.) Merrill) leaves to several X-ray doses through a polychromatic benchtop microprobe X-ray fluorescence spectrometer, modulating the photon flux by adjusting either the beam size, focus, or exposure time. The structure, ultrastructure and physiological responses of the irradiated plant tissues were investigated through light and transmission electron microscopy (TEM). Depending on the dose, the X-ray exposure induced decreased K and X-ray scattering intensities, and increased Ca, P, and Mn signals on soybean leaves. Anatomical analysis indicated necrosis of the epidermal and mesophyll cells on the irradiated spots, where TEM images revealed the collapse of cytoplasm and cell-wall breaking. Furthermore, the histochemical analysis detected the production of reactive oxygen species, as well as inhibition of chlorophyll autofluorescence in these areas. Under certain X-ray exposure conditions, e.g., high photon flux and exposure time, XRF measurements may affect the soybean leaves structures, elemental composition, and cellular ultrastructure, and induce programmed cell death. These results shed light on the characterization of the radiation damage, and thus, help to assess the X-ray radiation limits and strategies for in vivo for XRF analysis. HighlightBy exposing soybean leaves to several X-ray doses, we show that the characteristic X-ray induced elemental changes stem from plants physiological signalling or responses rather than only sample dehydration.
Why it matches plant phenotyping methods植物組織のin vivo XRF測定における放射線損傷と測定アーティファクトを評価し、適用限界と測定条件を検証する研究であり、フェノタイピング手法の技術的妥当性が中心です。
abstractX-ray fluorescence spectroscopy (XRF) is a powerful technique for the in vivo assessment of plant tissues.
Reproduction assets foundThe paper's DATA AVAILABILITY section states the raw data (XRF spectra/maps and imaging measurements) are fully available on Figshare at the authors' public DOI, which matches an allowed URL.Dataset · publicThe raw data herein presented is fully available at Figshare
repository: https://doi.org/10.6084/m9.figshare.1858438Open asset ↗Figshare · 10.6084/m9.figshare.1858438pdf-page:6 lines:1-93Code / dataset availability confirmedEurope PMC · bioRxiv · checked 15 Sept 2026
Biological systems are the sum of their dynamic 3-dimensional (3D) parts. Therefore, it is critical to study biological structures in 3D and at high resolutions to gain insights into their physiological functions. Electron microscopy of metal replicas of unroofed cells and isolated organelles has been a key technique to visualize intracellular structures at nanometer resolution. However, many of these protocols require specialized equipment and personnel to complete them. Here we present novel accessible protocols to analyze biological structures in unroofed cells and biochemically isolated organelles in 3D and at nanometer resolutions, focusing on Arabidopsis clathrin-coated vesicles (CCVs) - an essential trafficking organelle lacking detailed structural characterization due to their low preservation in classical electron microscopy techniques. First, we establish a protocol to visualize CCVs in unroofed cells using scanning-transmission electron microscopy (STEM) tomography, providing sufficient resolution to define the clathrin coat arrangements. Critically, the samples are prepared directly on electron microscopy grids, removing the requirement to use extremely corrosive acids, thereby enabling the use of this protocol in any electron microscopy lab. Secondly, we demonstrate this standardized sample preparation allows the direct comparison of isolated CCV samples with those visualized in cells. Finally, to facilitate the high-throughput and robust screening of metal replicated samples, we provide a deep learning analysis workflow to screen the ‘pseudo 3D’ morphology of CCVs imaged with 2D modalities. Overall, we present accessible ways to examine the 3D structure of biological samples and provide novel insights into the structure of plant CCVs.
Why it matches plant phenotyping methods植物細胞内オルガネラの3D形態を取得・解析する電子顕微鏡プロトコルと深層学習ワークフローが研究の中心であり、植物CCV形態の技術的スクリーニング手法を提供している。
abstractHere we present novel accessible protocols to analyze biological structures in unroofed cells and biochemically isolated organelles in 3D and at nanometer resolutions, focusing on Arabidopsis clathrin-coated vesicles (CCVs)
Reproduction assets foundThe paper's Data Availability statement explicitly deposits example data (SEM replica images, training image pairs) and the analysis code (Cellpose-based CCV segmentation workflow) generated in this study at a public Zenodo DOI, making it a paper-specific, publicly actionable asset. The temography.com URLs are vendor/mCode · publicand round; LF, large and
354
flat) using an area threshold of 8500 nm2
(a CCV diameter of 105 nm) and a 3D value of 1.52 (the
355
average of the 3 smallest CCVs in control conditions determined to be spherical by the experimenter).
356
Data Availability
357
Example data and the code generated in this study is available at:
358
https://doi.org/10.5281/zenodo.6563819
359
Acknowledgements
360
This research was supported by the Scientific Service Units of Institute of Science and Technology
361
Austria (ISTA) through resources provided by the Electron Microscopy Facility, Lab Support Facility and
362
the Imaging and Optics Facility. A.J. is supported by funding from the Austrian Science FundOpen asset ↗zenodo · 10.5281/zenodo.6563819pdf-raw-page:12 lines:1-46Code / dataset availability confirmedOpenAlex · Europe PMC · checked 14 Sept 2026
BACKGROUND: Although quantitative single-cell analysis is frequently applied in animal systems, e.g. to identify novel drugs, similar applications on plant single cells are largely missing. We have exploited the applicability of high-throughput microscopic image analysis on plant single cells using tobacco leaf protoplasts, cell-wall free single cells isolated by lytic digestion. Protoplasts regenerate their cell wall within several days after isolation and have the potential to expand and proliferate, generating microcalli and finally whole plants after the application of suitable regeneration conditions. RESULTS: High-throughput automated microscopy coupled with the development of image processing pipelines allowed to quantify various developmental properties of thousands of protoplasts during the initial days following cultivation by immobilization in multi-well-plates. The focus on early protoplast responses allowed to study cell expansion prior to the initiation of proliferation and without the effects of shape-compromising cell walls. We compared growth parameters of wild-type tobacco cells with cells expressing the antiapoptotic protein Bcl2-associated athanogene 4 from Arabidopsis (AtBAG4). CONCLUSIONS: AtBAG4-expressing protoplasts showed a higher proportion of cells responding with positive area increases than the wild type and showed increased growth rates as well as increased proliferation rates upon continued cultivation. These features are associated with reported observations on a BAG4-mediated increased resilience to various stress responses and improved cellular survival rates following transformation approaches. Moreover, our single-cell expansion results suggest a BAG4-mediated, cell-independent increase of potassium channel abundance which was hitherto reported for guard cells only. The possibility to explain plant phenotypes with single-cell properties, extracted with the single-cell processing and analysis pipeline developed, allows to envision novel biotechnological screening strategies able to determine improved plant properties via single-cell analysis.
Why it matches plant phenotyping methods植物プロトプラストの成長・増殖特性を大量画像から抽出する自動顕微鏡解析と画像処理パイプラインを開発・適用しており、表現型取得手法が研究の中心である。
abstractHigh-throughput automated microscopy coupled with the development of image processing pipelines allowed to quantify various developmental properties of thousands of protoplasts
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Code · publicAll the scripts and codes used in the analysis pipeline, additional downloaded plugins used in processing the images as well as sample data can be found in our Github page https://github.com/jodawson/cell_seg_tracking_analysis .Open asset ↗jodawson/cell_seg_tracking_analysislines:143-159Code / dataset availability confirmedOpenAlex · Crossref · Europe PMC · checked 8 Sept 2026
Confocal imaging is a well-established method for investigating plant phenotypes on the tissue and organ level. However, many differences are difficult to assess by visual inspection and researchers rely extensively on ad hoc manual quantification techniques and qualitative assessment. Here we present a method for quantitatively phenotyping large samples of plant tissue morphologies using triangulated isosurfaces. We successfully demonstrate the applicability of the approach using confocal imaging of aerial organs in Arabidopsis thaliana. Automatic identification of flower primordia using the surface curvature as an indication of outgrowth allows for high-throughput quantification of divergence angles and further analysis of individual flowers. We demonstrate the throughput of our method by quantifying geometric features of 1065 flower primordia from 172 plants, comparing auxin transport mutants to wild type. Additionally, we find that a paraboloid provides a simple geometric parameterisation of the shoot inflorescence domain with few parameters. We utilise parameterisation methods to provide a computational comparison of the shoot apex defined by a fluorescent reporter of the central zone marker gene CLAVATA3 with the apex defined by the paraboloid. Finally, we analyse the impact of mutations which alter mechanical properties on inflorescence dome curvature and compare the results with auxin transport mutants. Our results suggest that region-specific expression domains of genes regulating cell wall biosynthesis and local auxin transport can be important in maintaining the wildtype tissue shape. Altogether, our results indicate a general approach to parameterise and quantify plant development in 3D, which is applicable also in cases where data resolution is limited, and cell segmentation not possible. This enables researchers to address fundamental questions of plant development by quantitative phenotyping with high throughput, consistency and reproducibility.
Why it matches plant phenotyping methods植物組織の3D画像から形態形質を自動抽出・定量する手法を開発し、高スループット性と再現性を実証しているため、フェノタイピング手法が中心である。
abstractHere we present a method for quantitatively phenotyping large samples of plant tissue morphologies using triangulated isosurfaces.
Reproduction assets foundThe paper's data availability statement explicitly deposits all original source data (confocal phenotyping data of Arabidopsis shoot apical meristems) in the Cambridge Apollo repository and all analysis/segmentation/quantification scripts in a public Sainsbury Laboratory GitLab repository. Both are paper-specific,公开,直接Dataset · publicAll original source data files used in this study are available via the Cambridge University Apollo Repository ( https://doi.org/10.17863/CAM.82442 ).Open asset ↗Cambridge University Apollo Repository · 10.17863/CAM.82442lines:369-397Code · publicAll scripts and software for segmentation, quantification, analysis and visualisation are available via the Sainsbury Laboratory GitLab repository ( https://gitlab.com/slcu/teamHJ/publications/aahl_etal_2022 ).Open asset ↗Sainsbury Laboratory GitLab repositorylines:369-397Code / dataset availability confirmedEurope PMC · checked 8 Sept 2026
Ratiometric time-lapse FRET analysis requires a robust and accurate processing pipeline to eliminate bias in intensity measurements on fluorescent images before further quantitative analysis can be conducted. This level of robustness can only be achieved by supplementing automated tools with built-in flexibility for manual ad-hoc adjustments. FRET-IBRA is a modular and fully parallelized configuration file-based tool written in Python. It simplifies the FRET processing pipeline to achieve accurate, registered, and unified ratio image stacks. The flexibility of this tool to handle discontinuous image frame sequences with tailored configuration parameters further streamlines the processing of outliers and time-varying effects in the original microscopy images. FRET-IBRA offers cluster-based channel background subtraction, photobleaching correction, and ratio image construction in an all-in-one solution without the need for multiple applications, image format conversions, and/or plug-ins. The package accepts a variety of input formats and outputs TIFF image stacks along with performance measures to detect both the quality and failure of the background subtraction algorithm on a per frame basis. Furthermore, FRET-IBRA outputs images with superior signal-to-noise ratio and accuracy in comparison to existing background subtraction solutions, whilst maintaining a fast runtime. We have used the FRET-IBRA package extensively to quantify the spatial distribution of calcium ions during pollen tube growth under mechanical constraints. Benchmarks against existing tools clearly demonstrate the need for FRET-IBRA in extracting reliable insights from FRET microscopy images of dynamic physiological processes at high spatial and temporal resolution. The source code for Linux and Mac operating systems is released under the BSD license and, along with installation instructions, test images, example configuration files, and a step-by-step tutorial, is freely available at github.com/gmunglani/fret-ibra.
Why it matches plant phenotyping methods植物の動的な生理状態を画像から定量化するFRET画像処理ツールの開発・ベンチマークが中心であり、花粉管内カルシウム分布の抽出に実質的に応用されている。
abstractFRET-IBRA is a modular and fully parallelized configuration file-based tool written in Python.
Reproduction assets foundThe paper's authors publicly released the FRET-IBRA analysis toolkit (Python source code, test images, example configuration files, and tutorial) under a BSD license on GitHub. The test images include the FRET microscopy image stacks of growing Arabidopsis pollen tubes used in the paper's calcium-distribution phenotypiCode · publicThe source code for Linux and Mac operating systems is released under the BSD license and, along with installation instructions, test images, example configuration files, and a step-by-step tutorial, is freely available at github.com/gmunglani/fret-ibra.Open asset ↗github.com/gmunglani/fret-ibralines:113-128Code / dataset availability confirmedOpenAlex · Europe PMC · bioRxiv · checked 8 Sept 2026
Abstract In the absence of pollination, female reproductive organs senesce leading to an irrevocable loss in the reproductive potential of the flower and directly affecting seed set. In self-pollinating crops like wheat ( Triticum aestivum ), the post-anthesis viability of the unpollinated carpel has been overlooked, despite its importance for hybrid seed production systems. To advance our knowledge of carpel development in the absence of pollination, we created a relatively high-throughput phenotyping approach to quantify stigma and ovary morphology. We demonstrate the suitability of the approach, which is based on light microscopy imaging and machine learning, for the detailed study of floral organ traits in field grown plants using both fresh and fixed samples. We show that the unpollinated carpel undergoes a well-defined initial growth phase, followed by a peak phase (in which stigma area reaches its maximum and the radial expansion of the ovary slows), and a final deterioration phase. These developmental dynamics were largely consistent across years and could be used to classify male sterile cultivars, however the absolute duration of each phase varied across years. This phenotyping approach provides a new tool for examining carpel morphology and development which we hope will help advance research into this field and increase our mechanistic understanding of female fertility in wheat.
Why it matches plant phenotyping methodsコムギの柱頭・子房形態を定量化する高スループット表現型解析法を、光学顕微鏡画像と機械学習で開発・適用しており、表現型取得手法が研究の中心である。
abstractwe created a relatively high-throughput phenotyping approach to quantify stigma and ovary morphology.
Reproduction assets foundThe paper explicitly states that implementation scripts, data, and the trained stigma/ovary CNNs are publicly available at the authors' GitHub repository, which is an allowed URL.Code · publicImplementation scripts and data are available at
https://github.com/marina-millan/ML-carpel_traits.Open asset ↗marina-millan/ML-carpel_traits · ML-carpel_traitspdf-page:4 lines:1-61Code / dataset availability confirmedEurope PMC · Crossref · checked 13 Sept 2026
Arabidopsis (Arabidopsis thaliana) primary and lateral roots (LRs) are well suited for 3D and 4D microscopy, and their development provides an ideal system for studying morphogenesis and cell proliferation dynamics. With fast-advancing microscopy techniques used for live-imaging, whole tissue data are increasingly available, yet present the great challenge of analyzing complex interactions within cell populations. We developed a plugin "Live Plant Cell Tracking" (LiPlaCeT) coupled to the publicly available ImageJ image analysis program and generated a pipeline that allows, with the aid of LiPlaCeT, 4D cell tracking and lineage analysis of populations of dividing and growing cells. The LiPlaCeT plugin contains ad hoc ergonomic curating tools, making it very simple to use for manual cell tracking, especially when the signal-to-noise ratio of images is low or variable in time or 3D space and when automated methods may fail. Performing time-lapse experiments and using cell-tracking data extracted with the assistance of LiPlaCeT, we accomplished deep analyses of cell proliferation and clonal relations in the whole developing LR primordia and constructed genealogical trees. We also used cell-tracking data for endodermis cells of the root apical meristem (RAM) and performed automated analyses of cell population dynamics using ParaView software (also publicly available). Using the RAM as an example, we also showed how LiPlaCeT can be used to generate information at the whole-tissue level regarding cell length, cell position, cell growth rate, cell displacement rate, and proliferation activity. The pipeline will be useful in live-imaging studies of roots and other plant organs to understand complex interactions within proliferating and growing cell populations. The plugin includes a step-by-step user manual and a dataset example that are available at https://www.ibt.unam.mx/documentos/diversos/LiPlaCeT.zip.
Why it matches plant phenotyping methods植物の4Dライブイメージングから細胞系譜・位置・長さ・成長率などの形態・成長表現型を抽出する解析プラグインとパイプラインの開発が中心である。
abstractWe developed a plugin "Live Plant Cell Tracking" (LiPlaCeT) coupled to the publicly available ImageJ image analysis program and generated a pipeline that allows, with the aid of LiPlaCeT, 4D cell tracking and lineage analysis of populations of dividing and growing cells.
Reproduction assets foundThe paper's LiPlaCeT Fiji plugin for 4D plant cell tracking is publicly available: source code on GitHub and an ImageJ plugin package including a dataset example and user manual on the authors' IBT-UNAM site.Code · publicThe source code is freely available at https://github.com/paul-hernandez-herrera/LiPlaCeT and the ImageJ plugin including a dataset example and the User Manual can be downloaded from https://www.ibt.unam.mx/documentos/diversos/LiPlaCeT.zip .Open asset ↗paul-hernandez-herrera/LiPlaCeTlines:203-225Code / dataset availability confirmedEurope PMC · Crossref · checked 15 Sept 2026
Capturing complete internal anatomies of plant organs and tissues within their relevant morphological context remains a key challenge in plant science. While plant growth and development are inherently multiscale, conventional light, fluorescence, and electron microscopy platforms are typically limited to imaging of plant microstructure from small flat samples that lack a direct spatial context to, and represent only a small portion of, the relevant plant macrostructures. We demonstrate technical advances with a lab-based X-ray microscope (XRM) that bridge the imaging gap by providing multiscale high-resolution three-dimensional (3D) volumes of intact plant samples from the cell to the whole plant level. Serial imaging of a single sample is shown to provide sub-micron 3D volumes co-registered with lower magnification scans for explicit contextual reference. High-quality 3D volume data from our enhanced methods facilitate sophisticated and effective computational segmentation. Advances in sample preparation make multimodal correlative imaging workflows possible, where a single resin-embedded plant sample is scanned via XRM to generate a 3D cell-level map, and then used to identify and zoom in on sub-cellular regions of interest for high-resolution scanning electron microscopy. In total, we present the methodologies for use of XRM in the multiscale and multimodal analysis of 3D plant features using numerous economically and scientifically important plant systems.
Why it matches plant phenotyping methods植物の細胞から個体レベルの3D形態を取得するX線顕微鏡法と試料調製・計算セグメンテーションを中心に開発・提示しており、植物表現型取得手法が明確に主題である。
abstractWe demonstrate technical advances with a lab-based X-ray microscope (XRM) that bridge the imaging gap by providing multiscale high-resolution three-dimensional (3D) volumes of intact plant samples from the cell to the whole plant level.
Reproduction assets foundThe authors deposited fly-through animations of 2D image stacks and 3D volume rendering animations of the XRM scans shown in the paper's figures on figshare, directly reproducing this paper's plant phenotyping imaging data. No author analysis code or trained model checkpoints were explicitly deposited.Dataset · publicCanada) was used for data integration, visualization, and animation of the scan data, and to export image data as 2D 16-bit Tag Image File Format (TIFF) stacks. Fly-through animations of 2D image stacks for scans shown in all Figures, as well as 3D volume rendering animations of selected scans, are available for download from ( https://figshare.com/s/944efc8832e47fd4f203 ).
Image analysis and segmentation
Data from XRM scans were segmented using Amira software and with the assistance of a Wacom tablet for manual segmentation, in addition to ORS Dragonfly Deep Learning Module 2021.1.0.977 which is free for noncommercial use. Segmentation for Figure 1D combined automated and manual methods in Open asset ↗figsharelines:87-114Code / dataset availability confirmedOpenAlex · Europe PMC · checked 8 Sept 2026
Abstract Background Stalk lodging (mechanical failure of plant stems during windstorms) leads to global yield losses in cereal crops estimated to range from 5% to 25% annually. The cross-sectional morphology of plant stalks is a key determinant of stalk lodging resistance. However, previously developed techniques for quantifying cross-sectional morphology of plant stalks are relatively low-throughput, expensive and often require specialized equipment and expertise. There is need for a simple and cost-effective technique to quantify plant traits related to stalk lodging resistance in a high-throughput manner. Results A new phenotyping methodology was developed and applied to a range of plant samples including, maize ( Zea mays ), sorghum ( Sorghum bicolor ), wheat ( Triticum aestivum ), poison hemlock ( Conium maculatum ), and Arabidopsis (Arabis thaliana). The major diameter, minor diameter, rind thickness and number of vascular bundles were quantified for each of these plant types. Linear correlation analyses demonstrated strong agreement between the newly developed method and more time-consuming manual techniques (R 2 > 0.9). In addition, the new method was used to generate several specimen-specific finite element models of plant stalks. All the models compiled without issue and were successfully imported into finite element software for analysis. All the models demonstrated reasonable and stable solutions when subjected to realistic applied loads. Conclusions A rapid, low-cost, and user-friendly phenotyping methodology was developed to quantify two-dimensional plant cross-sections. The methodology offers reduced sample preparation time and cost as compared to previously developed techniques. The new methodology employs a stereoscope and a semi-automated image processing algorithm. The algorithm can be used to produce specimen-specific, dimensionally accurate computational models (including finite element models) of plant stalks.
Why it matches plant phenotyping methods植物茎の横断面形態を高スループットに定量する画像ベースの表現型計測法を開発し、手作業法との一致性検証と有限要素モデルへの応用を行っており、方法が研究の中心である。
abstractA new phenotyping methodology was developed and applied to a range of plant samples
Reproduction assets foundThe paper's MATLAB image-processing algorithm (authors' analysis code) and sample cross-sectional images are publicly available as supplementary files (Additional files 2 and 3) attached to this open-access article, along with standard operating protocols (Additional file 1). These directly reproduce the paper's phenotCode · publicThe code for the image-processing algorithm is also provided as Additional file 2 . Sample images and instructions are provided as Additional file 3 .Open asset ↗lines:110-119Code / dataset availability confirmedEurope PMC · checked 8 Sept 2026
Bicontinuous membranes in cell organelles epitomize nature's ability to create complex functional nanostructures. Like their synthetic counterparts, these membranes are characterized by continuous membrane sheets draped onto topologically complex saddle-shaped surfaces with a periodic network-like structure. Their structure sizes, (around 50-500 nm), and fluid nature make transmission electron microscopy (TEM) the analysis method of choice to decipher their nanostructural features. Here we present a tool, Surface Projection Image Recognition Environment (SPIRE), to identify bicontinuous structures from TEM sections through interactive identification by comparison to mathematical "nodal surface" models. The prolamellar body (PLB) of plant etioplasts is a bicontinuous membrane structure with a key physiological role in chloroplast biogenesis. However, the determination of its spatial structural features has been held back by the lack of tools enabling the identification and quantitative analysis of symmetric membrane conformations. Using our SPIRE tool, we achieved a robust identification of the bicontinuous diamond surface as the dominant PLB geometry in angiosperm etioplasts in contrast to earlier long-standing assertions in the literature. Our data also provide insights into membrane storage capacities of PLBs with different volume proportions and hint at the limited role of a plastid ribosome localization directly inside the PLB grid for its proper functioning. This represents an important step in understanding their as yet elusive structure-function relationship.
Why it matches plant phenotyping methods植物エチオプラストの膜構造をTEM画像から同定・定量解析するソフトウェアを開発し、植物器官の構造形質を抽出しているため、フェノタイピング手法が中心である。
abstractHere we present a tool, Surface Projection Image Recognition Environment (SPIRE), to identify bicontinuous structures from TEM sections through interactive identification by comparison to mathematical "nodal surface" models.
Reproduction assets foundThe paper's computational analysis tool SPIRE (used to identify and quantify prolamellar body cubic membrane structures from TEM micrographs) is explicitly released as open-source code with public URLs (SourceForge project and GitHub source repository), plus a video tutorial hosted at chloroplast.pl. No public deposit,Code · publicThe tool ( https://sourceforge.net/projects/spire-tool/ ) as well as the source code ( https://github.com/tohain/SPIRE ) and all dependencies are open source and thus freely and openly available.Open asset ↗spire-toollines:134-144Code · publicThe tool ( https://sourceforge.net/projects/spire-tool/ ) as well as the source code ( https://github.com/tohain/SPIRE ) and all dependencies are open source and thus freely and openly available.Open asset ↗SPIRElines:134-144Code / dataset availability confirmedEurope PMC · checked 15 Sept 2026
In plants, secretion of cell wall components and membrane proteins plays a fundamental role in growth and development as well as survival in diverse environments. Exocytosis, as the last step of the secretory trafficking pathway, is a highly ordered and precisely controlled process involving tethering, docking, and fusion of vesicles at the plasma membrane (PM) for cargo delivery. Although the exocytic process and machinery are well characterized in yeast and animal models, the molecular players and specific molecular events that underpin late stages of exocytosis in plant cells remain largely unknown. Here, by using the delivery of functional, fluorescent-tagged cellulose synthase (CESA) complexes (CSCs) to the PM as a model system for secretion, as well as single-particle tracking in living cells, we describe a quantitative approach for measuring the frequency of vesicle tethering events. Genetic and pharmacological inhibition of cytoskeletal function, reveal that the initial vesicle tethering step of exocytosis is dependent on actin and myosin XI. In contrast, treatments with the microtubule inhibitor, oryzalin, did not significantly affect vesicle tethering or fusion during CSC exocytosis but caused a minor increase in transient or aborted tethering events. With data from this new quantitative approach and improved spatiotemporal resolution of single particle events during secretion, we generate a revised model for the role of the cortical cytoskeleton in CSC trafficking.
Why it matches plant phenotyping methods植物細胞内の小胞テザリング頻度を単一粒子追跡で定量する新しい測定手法を開発・適用しており、植物状態の取得・定量が研究の中心である。
abstractsingle-particle tracking in living cells, we describe a quantitative approach for measuring the frequency of vesicle tethering events.
Reproduction assets foundThe paper's supplementary materials include Video S1, a live-cell imaging movie of a CSC particle insertion event next to a cortical microtubule, which directly reproduces the paper's plant phenotyping (single-particle CSC trafficking) measurements. No author analysis code or datasets with explicit deposit language areSupplement · publicnt care and maintenance of plant materials and to all members of the Staiger laboratory for helpful discussions and input.
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Supplementary Materials
The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/ijms23010317/s1 , Video S1: A CSC particle is inserted next to a cortical microtubule and translocates on the microtubule during the steady movement phase.
Click here for additional data file.
Author Contributions
W.Z. and C.J.S. designed the research. W.Z. performed the experiments and analyzed the data. W.Z. and C.J.S. wrote the Open asset ↗lines:64-115Code / dataset availability confirmedEurope PMC · OpenAlex · checked 15 Sept 2026
The three-dimensional (3D) arrangement of cells in tissues provides an anatomical basis for analyzing physiological and biochemical aspects of plant and animal cellular development and function. In this study, we established a protocol for tissue clearing and 3D imaging in rice. Our protocol is based on three improvements: clearing with iTOMEI (clearing solution suitable for plants), developing microscopic conditions in which the Z step is optimized for 3D reconstruction, and optimizing cell-wall staining. Our protocol successfully 3D imaged rice shoot apical meristems, florets, and root apical meristems at cellular resolution throughout whole tissues. Using fluorescent reporters of auxin signaling in rice root tips, we also revealed the 3D distribution of auxin signaling events that are activated in the columella, quiescent center, and multiple rows of cells in the stele of the root apical meristem. Examination of cells with higher levels of auxin signaling revealed that only the central row of cells was connected to the quiescent center. Our method provides opportunities to observe the 3D arrangement of cells in rice tissues.
Why it matches plant phenotyping methodsイネ組織を対象に、組織透明化・最適化した3D顕微鏡撮像・細胞壁染色による細胞配置の取得法を開発しており、植物表現型の画像取得が中心的な技術貢献である。
abstractIn this study, we established a protocol for tissue clearing and 3D imaging in rice.
Reproduction assets foundThe paper's 3D imaging datasets (supplementary videos S1–S6 of rice SAMs, florets, anthers, and root tips, plus figure data) are publicly available via the MDPI supplementary materials link. No separate analysis code repository is mentioned.Dataset · publiccquisition, which took approximately 2 h for 150 μm in depth, the images were processed using LASX software (Leica Microsystems, Tokyo, Japan).
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Materials
The following are available online at https://www.mdpi.com/article/10.3390/ijms23010040/s1 .
Click here for additional data file.
Author Contributions
M.S. and H.T. designed the research; M.S., H.A., Y.S. and S.M. performed the research; M.S. and H.T. analyzed the data; M.S. and H.T. wrote the paper. All authors have read and agreed to the published version of the manuscript.
Funding
This study was supporOpen asset ↗lines:70-202Code / dataset availability confirmedEurope PMC · checked 15 Sept 2026
Stomatal observation and automatic stomatal detection are useful analyses of stomata for taxonomic, biological, physiological, and eco-physiological studies. We present a new clearing method for improved microscopic imaging of stomata in soybean followed by automated stomatal detection by deep learning. We tested eight clearing agent formulations based upon different ethanol and sodium hypochlorite (NaOCl) concentrations in order to improve the transparency in leaves. An optimal formulation-a 1:1 ( v / v ) mixture of 95% ethanol and NaOCl (6-14%)-produced better quality images of soybean stomata. Additionally, we evaluated fixatives and dehydrating agents and selected absolute ethanol for both fixation and dehydration. This is a good substitute for formaldehyde, which is more toxic to handle. Using imaging data from this clearing method, we developed an automatic stomatal detector using deep learning and improved a deep-learning algorithm that automatically analyzes stomata through an object detection model using YOLO. The YOLO deep-learning model successfully recognized stomata with high mAP (~0.99). A web-based interface is provided to apply the model of stomatal detection for any soybean data that makes use of the new clearing protocol.
Why it matches plant phenotyping methodsダイズ葉の気孔画像取得法と、気孔を自動検出・解析する深層学習手法を開発・評価しており、植物形質取得が研究の中心である。
abstractWe present a new clearing method for improved microscopic imaging of stomata in soybean followed by automated stomatal detection by deep learning.
Reproduction assets foundThe paper's soybean stomatal phenotype dataset (Table S2, manual vs. automatic stomatal density for 386 accessions) is publicly available via the MDPI supplement, and the trained YOLOv5 stomata-detection model is publicly served through the authors' web application. The 183-image training dataset and analysis code haveSupplement · publicnowledge the personnel from the Plant Genetics and Breeding lab at the Kyungpook National University for their time and work at the greenhouse.
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Materials
The following are available online at https://www.mdpi.com/article/10.3390/plants10122714/s1 , Table S1: The representative methods for detecting stomata in different species, Table S2: Comparison stomata density between counting by manual and develped program.
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Author Contributions
Conceptualization, J.-D.L. and Y.J.K.; methodology, S.N.S.; formal analysis, H.P., S.H.COpen asset ↗lines:69-162Code / dataset availability confirmedCrossref · checked 9 Sept 2026
Significance The lack of suitable approaches for studying root–microbe interactions, live and in situ, has severely limited our ability to understand the rhizosphere. In this study, we overcome this major limitation with an imaging system that combines transparent soils with cutting edge light sheet microscopy. The study revealed that the root cap is a point of first contact for microbes before establishment and reveals how the pore structure influences the patterns of interactions between the microbe and the plant. With the combined use of light sheet microscopy and transparent soils, we shed light on previously unseen interaction phenomena and accelerate the understanding of how rhizospheres are formed.
Why it matches plant phenotyping methods透明土壌とライトシート顕微鏡を組み合わせたライブ・インサイチュ画像システムの開発が研究の中心で、根と微生物の相互作用という植物状態を可視化している。
abstractwe overcome this major limitation with an imaging system that combines transparent soils with cutting edge light sheet microscopy.
Reproduction assets foundThe paper deposits its phenotyping data (light-sheet microscopy volumes of root–soil–bacteria interactions) on Zenodo, makes its image analysis software (MATLAB/MeVisLab segmentation and quantification pipeline) publicly available on GitHub, and hosts supplementary materials on PNAS.Dataset · publicThe data in this study is available at https://doi.org/10.5281/zenodo.5650962 .Open asset ↗zenodo · 10.5281/zenodo.5650962lines:90-123Code · publicImage processing methods were programmed using MATLAB using the Image Processing Toolbox (MathWorks). Segmentation and extraction of geometrical features were performed using MeVisLab (MeVis Medical Solutions AG). All software is freely available from https://github.com/LionelDupuy/SENSOIL .Open asset ↗github · LionelDupuy/SENSOILlines:80-89Code / dataset availability confirmedEurope PMC · checked 15 Sept 2026
To measure stomatal traits automatically and nondestructively, a new method for detecting stomata and extracting stomatal traits was proposed. Two portable microscopes with different resolutions (TipScope with a 40× lens attached to a smartphone and ProScope HR2 with a 400× lens) are used to acquire images of living stomata in maize leaves. FPN model was used to detect stomata in the TipScope images and measure the stomata number and stomatal density. Faster RCNN model was used to detect opening and closing stomata in the ProScope HR2 images, and the number of opening and closing stomata was measured. An improved CV model was used to segment pores of opening stomata, and a total of 6 pore traits were measured. Compared to manual measurements, the square of the correlation coefficient (R 2 ) of the 6 pore traits was higher than 0.85, and the mean absolute percentage error (MAPE) of these traits was 0.02%-6.34%. The dynamic stomata changes between wild-type B73 and mutant Zmfab1a were explored under drought and re-watering condition. The results showed that Zmfab1a had a higher resilience than B73 on leaf stomata. In addition, the proposed method was tested to measure the leaf stomatal traits of other nine species. In conclusion, a portable and low-cost stomata phenotyping method that could accurately and dynamically measure the characteristic parameters of living stomata was developed. An open-access and user-friendly web portal was also developed which has the potential to be used in the stomata phenotyping of large populations in the future.
Why it matches plant phenotyping methods生きた葉の気孔形質を画像取得・深層学習・セグメンテーションで自動抽出する手法を開発し、手動測定との比較検証とWebポータル提供まで行っており、植物フェノタイピング手法が中心である。
abstracta new method for detecting stomata and extracting stomatal traits was proposed.
Reproduction assets foundThe paper's Data Availability Statement deposits the trained stomata detection/segmentation models and all labelled leaf stomata images at a public Huazhong Agricultural University plant phenomics download portal, which directly supports this paper's phenotyping measurements. The analysis source codes are only 'availabDataset · publical document of web portal.
File S2 Technical document of EXE software.
Data Availability Statement
The operating procedure for stomatal trait extraction is shown in Video S1 . The detailed technical documentation is given in Note S1 . The trained model, user guideline and all the labelled images of leaf stomata are available at http://plantphenomics.hzau.edu.cn/download_checkiflogin_en.action . The source codes are available from the first/corresponding author. The web portal of extracting stomatal traits was available at http://x40833180q.zicp.vip .Open asset ↗plantphenomics.hzau.edu.cnlines:638-643Code / dataset availability confirmedOpenAlex · Europe PMC · checked 14 Sept 2026
BACKGROUND: Cellular components are controlled by genetic and physiological factors that define their shape and size. However, quantitively capturing the morphological characteristics and movement of cellular organelles from micrograph images is challenging, because the analysis deals with complexities of images that frequently lead to inaccuracy in the estimation of the features. Here we show a unique quantitative method to overcome biases and inaccuracy of biological samples from confocal micrographs. RESULTS: We generated 2D images of cell walls and spindle-shaped cellular organelles, namely ER bodies, with a maximum contrast projection of 3D confocal fluorescent microscope images. The projected images were further processed and segmented by adaptive thresholding of the fluorescent levels in the cell walls. Micrographs are composed of pixels, which have information on position and intensity. From the pixel information we calculated three types of features (spatial, intensity and Haralick) in ER bodies corresponding to segmented cells. The spatial features include basic information on shape, e.g., surface area and perimeter. The intensity features include information on mean, standard deviation and quantile of fluorescence intensities within an ER body. Haralick features describe the texture features, which can be calculated mathematically from the interrelationship between the pixel information. Together these parameters were subjected to multivariate analysis to estimate the morphological diversity. Additionally, we calculated the displacement of the ER bodies using the positional information in time-lapse images. We captured similar morphological diversity and movement within ER body phenotypes in several microscopy experiments performed in different settings and scanned under different objectives. We then described differences in morphology and movement of ER bodies between A. thaliana wild type and mutants deficient in ER body-related genes. CONCLUSIONS: The findings unexpectedly revealed multiple genetic factors that are involved in the shape and size of ER bodies in A. thaliana. This is the first report showing morphological characteristics in addition to the movement of cellular components and it quantitatively summarises plant phenotypic differences even in plants that show similar cellular components. The estimation of morphological diversity was independent of the cell staining method and the objective lens used in the microscopy. Hence, our study enables a robust estimation of plant phenotypes by recognizing small differences in complex cell organelle shapes and their movement, which is beneficial in a comprehensive analysis of the molecular mechanism for cell organelle formation that is independent of technical variations.
Why it matches plant phenotyping methods植物細胞小器官の形態・移動を画像から抽出する特徴量計算法を開発し、異なる撮像条件で頑健性を検証しているため、表現型取得法が研究の中心です。
abstractHere we show a unique quantitative method to overcome biases and inaccuracy of biological samples from confocal micrographs.
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Code · publicThe z-stack images were merged using specific criteria for the MaxContrastProjection package ( https://github.com/arpankbasak/ERB_DynaMo ).Open asset ↗arpankbasak/ERB_DynaMolines:96-99Code / dataset availability confirmedEurope PMC · checked 15 Sept 2026
Plant stomata are essential structures (pores) that control the exchange of gases between plant leaves and the atmosphere, and also they influence plant adaptation to climate through photosynthesis and transpiration stream. Many works in literature aim for a better understanding of these structures and their role in the evolution process and the behavior of plants. Although stomata studies in dicots species have advanced considerably in the past years, even there is not much knowledge about the stomata of cereal grasses. Due to the high morphological variation of stomata traits intra- and inter-species, detecting and classifying stomata automatically becomes challenging. For this reason, in this work, we propose a new system for automatic stomata classification and detection in microscope images for maize cultivars based on transfer learning strategy of different deep convolution neural netwoks (DCNN). Our performed experiments show that our system achieves an approximated accuracy of 97.1% in identifying stomata regions using classifiers based on deep learning features, which figures out as a nearly perfect classification system. As the stomata are responsible for several plant functionalities, this work represents an important advance for maize research, providing an accurate system in replacing the current manual task of categorizing these pores on microscope images. Furthermore, this system can also be a reference for studies using images from different cereal grasses.
Why it matches plant phenotyping methodsトウモロコシ葉の気孔を顕微鏡画像から自動検出・分類する画像解析手法を開発し、精度評価しており、植物表現型取得が中心である。
abstractwe propose a new system for automatic stomata classification and detection in microscope images for maize cultivars based on transfer learning strategy of different deep convolution neural netwoks (DCNN).
Reproduction assets foundThe paper's Data Availability statement explicitly deposits all microscope images and analysis code in a public Zenodo record, directly reproducing this paper's maize stomata classification/detection experiments.Dataset · publicour findings can significantly benefit future research. As future work, we intend to develop a computational toolkit to support specialists in the biology area in their studies.
Supporting information
S1 File
(TXT)
Click here for additional data file.
Data Availability
All images and code are available from the ZENODO database. https://zenodo.org/record/3938047#.YE_l6v7Q85k .
Funding Statement
FAF received support of the Brazilian scientific funding agency CNPq through project #408919/2016-7 and São Paulo Research Foundation FAPESP grant #2018/23908-1. JPP received support of the Brazilian scientific funding agency CNPq through project #307066/2017-7. FAF received GPUs as donation from NVIDIOpen asset ↗ZENODO · zenodo.org/record/3938047lines:250-290Code / dataset availability confirmedEurope PMC · bioRxiv · checked 15 Sept 2026
Summary Mitochondria form highly dynamic populations in the cells of plants (and all eukaryotes). The characteristics of this collective behaviour, and how it is influenced by nuclear features, remain to be fully elucidated. Here, we use a recently-developed quantitative approach to reveal and analyse the physical and collective “social” dynamics of mitochondria in an Arabidopsis msh1 mutant where organelle DNA maintenance machinery is compromised. We use a newly-created line combining the msh1 mutant with mitochondrially-targeted GFP, and characterise mitochondrial dynamics with a combination of single-cell timelapse microscopy, computational tracking and network analysis. The collective physical behaviour of msh1 mitochondria is altered from wildtype in several ways: mitochondria become less evenly spread, and networks of inter-mitochondrial encounters become more connected with greater potential efficiency for inter-organelle exchange. We find that these changes are similar to those observed in friendly , where mitochondrial dynamics are altered by a physical perturbation, suggesting that this shift to higher connectivity may reflect a general response to mitochondrial challenges.
Why it matches plant phenotyping methods単なる生物学的測定ではなく、タイムラプス顕微鏡、計算追跡、ネットワーク解析を組み合わせて植物細胞内ミトコンドリアの動態状態を定量化する手法の実質的な適用である。
abstractwe use a recently-developed quantitative approach to reveal and analyse the physical and collective “social” dynamics of mitochondria
Reproduction assets foundThe paper explicitly states that all analysis code and data are available on the authors' GitHub repository, and a supplementary time-lapse microscopy video (phenotyping input) is hosted publicly. The Arabidopsis msh1 seed stock (N3372) used for the phenotyping is also publicly available from the NASC stock centre.Code · public14
average number of shortest paths crossing each node in the network. The mean connected
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component number is the average number of disconnected subgraphs within the network.
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Accession numbers
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All analysis code and data is available from Github at
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https://github.com/StochasticBiology/plant-mito-dynamics
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Acknowledgments
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J.M.C. is supported by the BBSRC and University of Birmingham via the MIBTP doctoral
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training scheme (grant number BB/M01116X/1). This project has received funding from the
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European Research Council (ERC) under the European Union’s Horizon 2020 research and
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innovation programme (grantOpen asset ↗StochasticBiology/plant-mito-dynamicspdf-raw-page:14 lines:1-67Code / dataset availability confirmedEurope PMC · OpenAlex · Crossref · checked 14 Sept 2026
The European mistletoe ( Viscum album ) is a dioecious epiphytic evergreen hemiparasite that develops an extensive endophyte enabling the absorption of water and mineral salts from the host tree, whereas the exophytic leaves are photosynthetically active. The attachment mode and host penetration are well studied, but little information is available about the effects of mistletoe age and sex on haustorium-host interactions. We harvested 130 plants of Viscum album ssp. album growing on host branches of Aesculus flava for morphological and anatomical investigations. Morphometric analyses of the mistletoe and the (hypertrophied) host interaction site were correlated with mistletoe age and sex. We recorded the morphology of the endophytic systems of various ages by using X-ray microtomography scans and corresponding stereomicroscopic images. For detailed anatomical studies, we examined thin stained sections of the mistletoe-host interface by light microscopy. The diameter and length of the branch hypertrophy showed a positive linear correlation with the age of the mistletoe. Correlations with their sex were only found for ratios between host branch and hypertrophy size. A female bias of about 76% was found. In a 4-year-old mistletoe, several small, almost equally sized sinkers and the connected cortical strands extend over more than 5 cm within the host branch. In older mistletoes, one main sinker was predominant and occupied an increasingly large proportion of the stem cross-section. Bands of vessels ran along the axis of the wedge-shaped haustoria and sinkers and bent sideways toward the mistletoe-host interface. At the interface, the vascular elements of the host wood changed their direction and formed vortices near the haustorium.
Why it matches plant phenotyping methodsマイクロCTとステレオ画像による植物内部構造の3D可視化・形態計測が研究の中心的手法であり、ミストルの内生系や宿主との相互作用部位という植物形態形質を取得している。
titleAdvances on the Visualization of the Internal Structures of the European Mistletoe: 3D Reconstruction Using Microtomography
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Supplement · publicSupplementary Table 1
Raw data of diameters and lengths of host branch (hypertrophy) and mistletoe.Open asset ↗lines:257-351Code / dataset availability confirmedOpenAlex · Europe PMC · checked 8 Sept 2026
The stomatal index of the leaf is the ratio of the number of stomata to the total number of stomata and epidermal cells. Comparing with the stomatal density, the stomatal index is relatively constant in environmental conditions and the age of the leaf and, therefore, of diagnostic characteristics for a given genotype or species. Traditional assessment methods involve manual counting of the number of stomata and epidermal cells in microphotographs, which is labor-intensive and time-consuming. Although several automatic measurement algorithms of stomatal density have been proposed, no stomatal index pipelines are currently available. The main aim of this research is to develop an automated stomatal index measurement pipeline. The proposed method employed Faster regions with convolutional neural networks (R-CNN) and U-Net and image-processing techniques to count stomata and epidermal cells, and subsequently calculate the stomatal index. To improve the labeling speed, a semi-automatic strategy was employed for epidermal cell annotation in each micrograph. Benchmarking the pipeline on 1,000 microscopic images of leaf epidermis in the wheat dataset (Triticum aestivum L.), the average counting accuracies of 98.03 and 95.03% for stomata and epidermal cells, respectively, and the final measurement accuracy of the stomatal index of 95.35% was achieved. R2 values between automatic and manual measurement of stomata, epidermal cells, and stomatal index were 0.995, 0.983, and 0.895, respectively. The average running time (ART) for the entire pipeline could be as short as 0.32 s per microphotograph. The proposed pipeline also achieved a good transferability on the other families of the plant using transfer learning, with the mean counting accuracies of 94.36 and 91.13% for stomata and epidermal cells and the stomatal index accuracy of 89.38% in seven families of the plant. The pipeline is an automatic, rapid, and accurate tool for the stomatal index measurement, enabling high-throughput phenotyping, and facilitating further understanding of the stomatal and epidermal development for the plant physiology community. To the best of our knowledge, this is the first deep learning-based microphotograph analysis pipeline for stomatal index assessment.
Why it matches plant phenotyping methods葉の顕微鏡画像から気孔と表皮細胞を検出・計数し、気孔指数を自動推定する画像解析パイプラインの開発とベンチマーク検証が研究の中心である。
abstractThe main aim of this research is to develop an automated stomatal index measurement pipeline.
Reproduction assets foundThe authors explicitly state the stomatal index pipeline code is fully open-source on GitHub and the wheat microscopic image dataset is downloadable as a zip release from the same repository. Both are paper-specific, public, and directly actionable.Code · publicThe code is fully open-source for academic usage and can be downloaded at https://github.com/WeizhenLiuBioinform/stomatal_indexOpen asset ↗WeizhenLiuBioinform/stomatal_indexlines:361-376Dataset · publicThe wheat dataset is available for downloading at https://github.com/WeizhenLiuBioinform/stomatal_index/releases/download/wheat1.0/wheat_dataset.zipOpen asset ↗WeizhenLiuBioinform/stomatal_index · wheat1.0lines:361-376Supplement · publicSupplementary Table 2
Description of the cuticle dataset used for training and testing the stomatal index measurement model.Open asset ↗lines:651-703Code / dataset availability confirmedCrossref · Europe PMC · checked 15 Sept 2026
Recent technical advances in the computer-vision domain have facilitated the development of various methods for achieving image-based quantification of stomata-related traits. However, the installation cost of such a system and the difficulties of operating it on-site have been hurdles for experimental biologists. Here, we present a platform that allows real-time stomata detection during microscopic observation. The proposed system consists of a deep neural network model-based stomata detector and an upright microscope connected to a USB camera and a graphics processing unit (GPU)-supported single-board computer. All the hardware components are commercially available at common electronic commerce stores at a reasonable price. Moreover, the machine-learning model is prepared based on freely available cloud services. This approach allows users to set up a phenotyping platform at low cost. As a proof of concept, we trained our model to detect dumbbell-shaped stomata from wheat leaf imprints. Using this platform, we collected a comprehensive range of stomatal phenotypes from wheat leaves. We confirmed notable differences in stomatal density ( SD ) between adaxial and abaxial surfaces and in stomatal size ( SS ) between wheat-related species of different ploidy. Utilizing such a platform is expected to accelerate research that involves all aspects of stomata phenotyping.
Why it matches plant phenotyping methods低コストの顕微鏡画像と深層学習による気孔検出・形質定量化プラットフォームの開発であり、植物フェノタイピング手法が研究の中心です。
abstractHere, we present a platform that allows real-time stomata detection during microscopic observation.
Reproduction assets foundThe paper's stomata-detection GUI, trained SSD model weights, and model-training workflow are publicly available in the authors' GitHub repository (onsite_stomata_platform) with an executable Colab training notebook. The raw phenotype/image datasets are only available on request per the Data Availability Statement.Code · publicDetailed codes and instructions to reproduce the regarding system as well as the stomata detection model is described in Google Colaboratory executable notebook 7 hosted at https://github.com/totti0223/onsite_stomata_platform .Open asset ↗totti0223/onsite_stomata_platformlines:155-166Code / dataset availability confirmedCrossref · checked 9 Sept 2026
Laboratory / benchtopChlorophyll fluorescenceMicroscopyRootGrowth / time-series analysisVisualization / data management
Fabricated ecosystems (EcoFABs) offer an innovative approach to in situ examination of microbial establishment patterns around plant roots using nondestructive, high-resolution microscopy. Previously high-resolution imaging was challenging because the roots were not constrained to a fixed distance from the objective. Here, we describe a new ‘Imaging EcoFAB’ and the use of this device to image the entire root system of growing Brachypodium distachyon at high resolutions (20×, 40×) over a 3-week period. The device is capable of investigating root–microbe interactions of multimember communities. We examined nine strains of Pseudomonas simiae with different fluorescent constructs to B. distachyon and individual cells on root hairs were visible. Succession in the rhizosphere using two different strains of P. simiae was examined, where the second addition was shown to be able to establish in the root tissue. The device was suitable for imaging with different solid media at high magnification, allowing for the imaging of fungal establishment in the rhizosphere. Overall, the Imaging EcoFAB could improve our ability to investigate the spatiotemporal dynamics of the rhizosphere, including studies of fluorescently-tagged, multimember, synthetic communities.
Why it matches plant phenotyping methods植物根系全体を高解像度・経時的に撮像するための新規チャンバーを開発し、その撮像性能と用途を示しており、表現型取得法が研究の中心である。
abstractHere, we describe a new ‘Imaging EcoFAB’ and the use of this device to image the entire root system of growing Brachypodium distachyon at high resolutions (20×, 40×) over a 3-week period.
Reproduction assets foundThe paper's computational analysis (K-means clustering and segmentation/cell counting of the 40× multispectral root image) is explicitly stated to have its environment, code, and parent data file available in the Supplementary Materials, hosted at the MDPI S1 link. Additionally, the 3D-printing-ready Imaging EcoFAB 3D-Code · publicThe environment, code, and parent data file are available in the Supplementary Materials .Open asset ↗lines:65-81Code / dataset availability confirmedbioRxiv · Europe PMC · checked 15 Sept 2026
Plants respond to the surrounding environment in countless ways. One of these responses is their ability to sense and orient their root growth toward the gravity vector. Root gravitropism is studied in many laboratories as a hallmark of auxin-related phenotypes. However, manual analysis of images and microscopy data is known to be subjected to human bias. This is particularly the case for manual measurements of root bending as the selection lines to calculate the angle are set subjectively. Therefore, it is essential to develop and use automated or semi-automated image analysis to produce reproducible and unbiased data. Moreover, the increasing usage of vertical-stage microscopy in plant root biology yields gravitropic experiments with an unprecedented spatiotemporal resolution. To this day, there is no available solution to measure root bending angle over time for vertical-stage microscopy. To address these problems, we developed ACORBA (Automatic Calculation Of Root Bending Angles), a fully automated software to measure root bending angle over time from vertical-stage microscope and flatbed scanner images. Moreover, the software can be used semi-automated for camera, mobile phone or stereomicroscope images. ACORBA represents a flexible approach based on both traditional image processing and deep machine learning segmentation to measure root angle progression over time. By its automated nature, the workflow is limiting human interactions and has high reproducibility. ACORBA will support the plant biologist community by reducing time and labor and by producing quality results from various kinds of inputs. Significance statementACORBA is implementing an automated and semi-automated workflow to quantify root bending and waving angles from images acquired with a microscope, a scanner, a stereomicroscope or a camera. It will support the plant biology community by reducing time and labor and by producing trustworthy and reproducible quantitative data.
Why it matches plant phenotyping methods根の屈曲角度を画像から自動抽出するソフトウェアとワークフローの開発が研究の中心であり、植物形態表現型の定量手法に該当する。
abstractwe developed ACORBA (Automatic Calculation Of Root Bending Angles), a fully automated software to measure root bending angle over time from vertical-stage microscope and flatbed scanner images.
Reproduction assets foundThe paper explicitly releases the ACORBA software (source code, trained models, annotated training libraries, notebooks, user manual) on SourceForge and the raw microscopy/scanner image stacks used for the root-angle measurements on Zenodo (DOI 10.5281/zenodo.5105719). Both are paper-specific, public, and actionable.Code · publicand
online Python image analysis and machine learning tutorials.
Availability of data and materials
The latest versions of ACORBA software training annotated libraries, source code, examples,
image pre-processing scripts, deep machine learning model training Jupyter notebooks and user
manual maintained by NBCS are available at https://sourceforge.net/projects/acorba/. The raw
microscopy and scanner stacks used in this paper are available at ZENODO
(https://doi.org/10.5281/zenodo.5105719). The analyzed results are supplemented (Supplemental
data).
Competing interests
The authors declare that they have no competing interests.
Funding
This work was supported by the European Research Council (GOpen asset ↗sourceforge.net/projects/acorbapdf-raw-page:17 lines:1-45Dataset · publicACORBA software training annotated libraries, source code, examples,
image pre-processing scripts, deep machine learning model training Jupyter notebooks and user
manual maintained by NBCS are available at https://sourceforge.net/projects/acorba/. The raw
microscopy and scanner stacks used in this paper are available at ZENODO
(https://doi.org/10.5281/zenodo.5105719). The analyzed results are supplemented (Supplemental
data).
Competing interests
The authors declare that they have no competing interests.
Funding
This work was supported by the European Research Council (Grant No. 803048), Charles
University Primus (Grant No. PRIMUS/19/SCI/09).
Author contributions
NBCS and MF conceived the pOpen asset ↗ZENODO · 10.5281/zenodo.5105719pdf-raw-page:17 lines:1-45Code / dataset availability confirmedEurope PMC · checked 9 Sept 2026
Ratiometric time-lapse FRET analysis requires a robust and accurate processing pipeline to eliminate bias in intensity measurements on fluorescent images before further quantitative analysis can be conducted. This level of robustness can only be achieved by supplementing automated tools with built-in flexibility for manual ad-hoc adjustments. FRET-IBRA is a modular and fully parallelized configuration file-based tool written in Python. It simplifies the FRET processing pipeline to achieve accurate, registered, and unified ratio image stacks. The flexibility of this tool to handle discontinuous image frame sequences with tailored configuration parameters further streamlines the processing of outliers and time-varying effects in the original microscopy images. FRET-IBRA offers cluster-based channel background subtraction, photobleaching correction, and ratio image construction in an all-in-one solution without the need for multiple applications, image format conversions, and/or plug-ins. The package accepts a variety of input formats and outputs TIFF image stacks along with performance measures to detect both the quality and failure of the background subtraction algorithm on a per frame basis. Furthermore, FRET-IBRA outputs images with superior signal-to-noise ratio and accuracy in comparison to existing background subtraction solutions, whilst maintaining a fast runtime. The FRET-IBRA package has been extensively used in quantifying the spatial distribution of calcium ions during pollen tube growth under mechanical constraints. Benchmarks against existing tools clearly demonstrate the need for FRET-IBRA in extracting reliable insights from FRET microscopy images of dynamic physiological processes at high spatial and temporal resolution. The source code for Linux and Mac operating systems is released under the BSD license and, along with installation instructions, test images, example configuration files, and a step-by-step tutorial, is freely available at github.com/gmunglani/fret-ibra . Author Summary FRET is a fundamental imaging technique used to generate fluorescence signals sensitive to molecular conformations and interactions. Despite its wide use and the large body of literature on the theoretical steps required to process images generated from this procedure, we were unable to locate a tool that contained the entire processing workflow, whilst allowing the user the flexibility to adjust parameters for maximum accuracy and runtime efficiency. FRET-IBRA was thus created to be an all-in-one, open-source, parallel solution to process FRET images, while eliminating complications arising from repeated image format conversions. Besides enhancing the background subtraction algorithm for FRET images, several additional options were implemented for the user to extract the cleanest signal possible for their specific use case. FRET-IBRA is primarily built for flexibility when handling large image stacks by supporting sequences of image frames to be treated independently, greatly reducing time spent on splitting and concatenating image stacks. In accuracy and speed benchmarks against more general background subtraction packages, FRET-IBRA was able to provide the cleanest results with a fast runtime, leading to reliable analysis without additional tuning.
Why it matches plant phenotyping methodsFRET画像の背景補正・補正処理・比画像構築を一体化したソフトウェアを開発し、既存ツールとのベンチマークで検証している。花粉管成長中のカルシウム分布という植物生理状態の抽出にも適用され、方法が中心である。
abstractFRET-IBRA is a modular and fully parallelized configuration file-based tool written in Python.
Reproduction assets foundThe paper's authors publicly release the FRET-IBRA analysis toolkit (source code, test images, example configuration files, and tutorial) on GitHub, directly supporting the paper's FRET image processing and ratiometric analysis of pollen tube calcium imaging.Code · publicThe source code for Linux and Mac operating systems is released under the
BSD license and, along with installation instructions, test images, example configuration
files, and a step-by-step tutorial, is freely available at github.com/gmunglani/fret-ibra.Open asset ↗github.com/gmunglani/fret-ibrapdf-page:1 lines:1-66Code / dataset availability confirmedbioRxiv · Europe PMC · checked 15 Sept 2026
Soil fungi establish mutualistic interactions with the roots of most vascular land plants. Arbuscular mycorrhizal (AM) fungi are among the most extensively characterised mycobionts to date. Current approaches to quantifying the extent of root colonisation and the abundance of hyphal structures in mutant roots rely on staining and human scoring involving simple, yet repetitive tasks prone to variations between experimenters. We developed AMFinder which allows for automatic computer vision-based identification and quantification of AM fungal colonisation and intraradical hyphal structures on ink-stained root images using convolutional neural networks. AMFinder delivered high-confidence predictions on image datasets of roots of multiple plant hosts (Nicotiana benthamiana, Medicago truncatula, Lotus japonicus, Oryza sativa) and captured the altered colonisation in ram1-1, str and smax1 mutants. A streamlined protocol for sample preparation and imaging allowed us to quantify mycobionts from the genera Rhizophagus, Claroideoglomus, Rhizoglomus and Funneliformis via flatbed scanning or digital microscopy including dynamic increases in colonisation in whole root systems over time. AMFinder adapts to a wide array of experimental conditions. It enables accurate, reproducible analyses of plant root systems and will support better documentation of AM fungal colonisation analyses. AMFinder can be accessed here: https://github.com/SchornacklabSLCU/amfinder.git
Why it matches plant phenotyping methods植物根の菌根菌感染状態を画像から自動識別・定量する手法とソフトウェアを開発しており、表現型取得・抽出が研究の中心です。
abstractWe developed AMFinder which allows for automatic computer vision-based identification and quantification of AM fungal colonisation and intraradical hyphal structures on ink-stained root images using convolutional neural networks.
Reproduction assets foundThe paper's AMFinder analysis software (amf/amfbrowser) and pre-trained CNN models are publicly available on the authors' GitHub repository under the MIT license. The training image datasets are not public and must be requested from the authors.Code · publicmanuscript. All authors
have read and approved the manuscript.
Data Availability
AMFinder is released under the terms of the open-source MIT license
(https://opensource.org/licenses/MIT) allowing unrestricted usage. Source code, pre-trained models
and detailed installation instructions are available on AMFinder GitHub webpage
(https://github.com/SchornacklabSLCU/amfinder.git). Training datasets are available upon request.
References
Abadi M, Agarwal A, Barham P, Brevdo E, Chen Z, Citro C, Corrado GS, Davis A, Dean J,
Devin M, et al. 2016. TensorFlow: Large-Scale Machine Learning on Heterogeneous Distributed
Systems. arXiv.
Bally J, Jung H, Mortimer C, Naim F, Philips JG, Hellens R, BombarelyOpen asset ↗SchornacklabSLCU/amfinderpdf-raw-page:21 lines:1-72Code / dataset availability confirmedOpenAlex · Europe PMC · checked 15 Sept 2026
Abstract Background: Cellular components are controlled by genetic and physiological factors that define their shape and size. However, quantitively capturing the morphological characteristics and movement of cellular organelles from micrograph images is challenging, because the analysis deals with complexities of images that frequently lead to inaccuracy in the estimation of the features. Here we show a unique quantitative method to overcome biases and inaccuracy of biological samples from confocal micrographs. Results: We generated 2D images of cell walls and spindle-shaped cellular organelles, namely ER bodies, with a maximum contrast projection of 3D confocal fluorescent microscope images. The projected images were further processed and segmented by adaptive thresholding of the fluorescent levels in the cell walls. Micrographs are composed of pixels, which have information on position and intensity. From the pixel information we calculated three types of features (spatial, intensity and Haralick) in ER bodies corresponding to segmented cells. The spatial features include basic information on shape, e.g., surface area and perimeter. The intensity features include information on mean, standard deviation and quantile of fluorescence intensities within an ER body. Haralick features describe the texture features, which can be calculated mathematically from the interrelationship between the pixel information. Together these parameters were subjected to multivariate analysis to estimate the morphological diversity. Additionally, we calculated the displacement of the ER bodies using the positional information in a time-lapse image. We captured similar morphological diversity and movement within ER body phenotypes on several microscopy experiments performed in different settings and scanned under different objectives. We then described differences in morphology and movement of ER bodies between A. thaliana wild type and mutants deficient in ER body-related genes. Conclusions: The findings unexpectedly revealed multiple genetic factors that are involved in the shape and size of ER bodies in A. thaliana . This is the first report showing morphological characteristics in addition to the movement of cellular components and quantitatively summarises plant phenotypic differences even in plants that show similar cellular components. The estimation of morphological diversity was independent of the cell staining method and the objective lens used in the microscopy. Hence, our study enables a robust estimation of plant phenotypes by recognizing small differences of complex cell organelle shapes and their movement, which is beneficial in a comprehensive analysis of the molecular mechanism for cell organelle formation that is independent of technical variations.
Why it matches plant phenotyping methods顕微鏡画像からERボディの形態・テクスチャ・移動を抽出し、異なる実験条件で頑健性を検証する植物表現型解析手法が中心である。
abstractHere we show a unique quantitative method to overcome biases and inaccuracy of biological samples from confocal micrographs.
Reproduction assets foundThe paper's authors publicly release their phenotyping analysis scripts (segmentation, feature extraction, dynamics, clustering) on GitHub and the conda analysis environment on Anaconda Cloud, both with explicit availability statements. Microscope images are said to be in a 'Bioimage database' but no URL is given, so aCode · publicell as the institutional core support by Małopolska Centre of Biotechnology, Jagiellonian University.
Availability of data and materials
The scripts used can be found in the GitHub repository, images can be found in the Bioimage database. Scripts used for pre-processing and analysis along with supplementary data can be found in http://www.github.com/arpankbasak/ERB_DynaMo The analysis environment erb_dynamo with all necessary dependencies have been uploaded to https://anaconda.org/arpankbasak/erb_dynamo/files . The plant materials are available from the Arabidopsis Biological Resource Center (ABRC) and Nottingham Arabidopsis Stock Center (NASC).
Ethics approval and consent to participate
NotOpen asset ↗arpankbasak/ERB_DynaMolines:342-369Code · publics
The scripts used can be found in the GitHub repository, images can be found in the Bioimage database. Scripts used for pre-processing and analysis along with supplementary data can be found in http://www.github.com/arpankbasak/ERB_DynaMo The analysis environment erb_dynamo with all necessary dependencies have been uploaded to https://anaconda.org/arpankbasak/erb_dynamo/files . The plant materials are available from the Arabidopsis Biological Resource Center (ABRC) and Nottingham Arabidopsis Stock Center (NASC).
Ethics approval and consent to participate
Not applicable.
Consent for publication
Consent and approval for publication from all the authors was obtained.
Competing Interests
The auOpen asset ↗arpankbasak/erb_dynamolines:342-369Code / dataset availability confirmedEurope PMC · checked 15 Sept 2026
Motivation Arbuscular mycorrhizas are the most widespread plant symbioses and involve the majority of crop plants. The beneficial interaction between plant roots and a group of soil fungi (Glomeromycotina) grants the green host a preferential access to soil mineral nutrients and water, supporting plant health, biomass production and resistance to both abiotic and biotic stresses. The nutritional exchanges at the core of this symbiosis take place inside the living root cells, which are diffusely colonized by specialized fungal structures called arbuscules. For this reason, the vast majority of studies investigating arbuscular mycorrhizas and their applications in agriculture require a precise quantification of the intensity of root colonization. To this aim, several manual methods have been used for decades to estimate the extension of intraradical fungal structures, mostly based on optical microscopy observations and individual assessment of fungal abundance in the root tissues. Results: Here we propose a novel semi-automated approach to quantify AM colonization based on digital image analysis and compare two methods based on image thresholding and machine learning. Our results indicate in machine learning a very promising tool for accelerating, simplifying and standardizing this critical type of analysis, with a direct potential interest for applicative and basic research. Contact ivan.sciascia@unito.it; andrea.genre@unito.it
Why it matches plant phenotyping methods植物根の菌根コロニー形成という植物状態を、画像解析・画像しきい値処理・機械学習で定量する手法の提案と比較検証が中心である。
abstractHere we propose a novel semi-automated approach to quantify AM colonization based on digital image analysis and compare two methods based on image thresholding and machine learning.
Reproduction assets foundThe authors deposited the microscopy image datasets used for binary, thresholding, and machine-learning segmentation of mycorrhizal Medicago truncatula roots in three public Figshare repositories, explicitly listed under 'Availability of data and material'. These are paper-specific phenotype image datasets directly支撑本.Dataset · public22
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Thresholding and machine learning segmentation - mycorrhized roots DOI
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Thresholding and machine learning segmentation – non mycorrhized roots DOI
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Thresholding and machine learning segmentation - mycorrhized roots DOI
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Thresholding and machine learning segmentation – non mycorrhized roots DOI
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Ministero dell’Istruzione, dell’Università e della Ricerca: PhD fellowship to AC UniveOpen asset ↗Figshare · 10.6084/m9.figshare.14679729pdf-raw-page:23 lines:1-85Dataset · publicavailable in the Figshare
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https://doi.org/10.6084/m9.figshare.14679729
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https://doi.org/10.6084/m9.figshare.14679684
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Authors' contributions
357
IS designed the image analysis approach, performed image anOpen asset ↗Figshare · 10.6084/m9.figshare.14679684pdf-raw-page:23 lines:1-85Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
Patchy stomata are a common and characteristic phenomenon in plants. Understanding and studying the regulation mechanism of patchy stomata are of great significance to further supplement and improve the stomatal theory. Currently, the common methods for stomatal behavior observation are based on static images, which makes it difficult to reflect dynamic changes of stomata. With the rapid development of portable microscopes and computer vision algorithms, it brings new chances for stomatal movement observation. In this study, a stomatal behavior observation system (SBOS) was proposed for real-time observation and automatic analysis of each single stoma in wheat leaf using object tracking and semantic segmentation methods. The SBOS includes two modules: the real-time observation module and the automatic analysis module. The real-time observation module can shoot videos of stomatal dynamic changes. In the automatic analysis module, object tracking locates every single stoma accurately to obtain stomatal pictures arranged in time-series; semantic segmentation can precisely quantify the stomatal opening area (SOA), with a mean pixel accuracy (MPA) of 0.8305 and a mean intersection over union (MIoU) of 0.5590 in the testing set. Moreover, we designed a graphical user interface (GUI) so that researchers could use this automatic analysis module smoothly. To verify the performance of the SBOS, the dynamic changes of stomata were observed and analyzed under chilling. Finally, we analyzed the correlation between gas exchange and SOA under drought stress, and the correlation coefficients between mean SOA and net photosynthetic rate (Pn), intercellular CO 2 concentration (Ci), stomatal conductance (Gs), and transpiration rate (Tr) are 0.93, 0.96, 0.96, and 0.97.
Why it matches plant phenotyping methods顕微鏡動画から個々の気孔を追跡し、セグメンテーションで気孔開口面積を定量化する観測・解析システムを開発しており、植物表現型取得が研究の中心です。
abstracta stomatal behavior observation system (SBOS) was proposed for real-time observation and automatic analysis of each single stoma in wheat leaf using object tracking and semantic segmentation methods.
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Code · public(2) The module is easy to install with the aid of an executable program (EXE) ( https://github.com/shem123456/Stomata-segmentation-with-GUI ).Open asset ↗https://github.com/shem123456/Stomata-segmentation-with-GUIlines:61-68Code / dataset availability confirmedEurope PMC · Crossref · OpenAlex · checked 14 Sept 2026
Parasitic plants infect other plants by forming haustoria, specialized multicellular organs consisting of several cell types, each of which has unique morphological features and physiological roles associated with parasitism. Understanding the spatial organization of cell types is, therefore, of great importance in elucidating the functions of haustoria. Here, we report a three-dimensional (3-D) reconstruction of haustoria from two Orobanchaceae species, the obligate parasite Striga hermonthica infecting rice (Oryza sativa) and the facultative parasite Phtheirospermum japonicum infecting Arabidopsis (Arabidopsis thaliana). In addition, field-emission scanning electron microscopy observation revealed the presence of various cell types in haustoria. Our images reveal the spatial arrangements of multiple cell types inside haustoria and their interaction with host roots. The 3-D internal structures of haustoria highlight differences between the two parasites, particularly at the xylem connection site with the host. Our study provides cellular and structural insights into haustoria of S. hermonthica and P. japonicum and lays the foundation for understanding haustorium function.
Why it matches plant phenotyping methods植物器官の3次元画像再構成を中心に、ハウストリアの内部構造と細胞配置を可視化しており、形態状態の取得・抽出が研究の主要部分である。
titleThree-dimensional reconstructions of haustoria in two parasitic plant species in the Orobanchaceae
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Code · publicAfter automated alignment adjustment, section alignment was manually checked and misaligned sections were re-registered by changing the registration parameters. The tools are available at https://github.com/yk-szk/ssrvtools .Open asset ↗yk-szk/ssrvtoolslines:86-98Code / dataset availability confirmedEurope PMC · checked 9 Sept 2026
Background Stomata analysis using microscope imagery provides important insight into plant physiology, health and the surrounding environmental conditions. Plant scientists are now able to conduct automated high-throughput analysis of stomata in microscope data, however, existing detection methods are sensitive to the appearance of stomata in the training images, thereby limiting general applicability. In addition, existing methods only generate bounding-boxes around detected stomata, which require users to implement additional image processing steps to study stomata morphology. In this paper, we develop a fully automated, robust stomata detection algorithm which can also identify individual stomata boundaries regardless of the plant species, sample collection method, imaging technique and magnification level. Results The proposed solution consists of three stages. First, the input image is pre-processed to remove any colour space biases occurring from different sample collection and imaging techniques. Then, a Mask R-CNN is applied to estimate individual stomata boundaries. The feature pyramid network embedded in the Mask R-CNN is utilised to identify stomata at different scales. Finally, a statistical filter is implemented at the Mask R-CNN output to reduce the number of false positive generated by the network. The algorithm was tested using 16 datasets from 12 sources, containing over 60,000 stomata. For the first time in this domain, the proposed solution was tested against 7 microscope datasets never seen by the algorithm to show the generalisability of the solution. Results indicated that the proposed approach can detect stomata with a precision, recall, and F-score of 95.10%, 83.34%, and 88.61%, respectively. A separate test conducted by comparing estimated stomata boundary values with manually measured data showed that the proposed method has an IoU score of 0.70; a 7% improvement over the bounding-box approach. Conclusions The proposed method shows robust performance across multiple microscope image datasets of different quality and scale. This generalised stomata detection algorithm allows plant scientists to conduct stomata analysis whilst eliminating the need to re-label and re-train for each new dataset. The open-source code shared with this project can be directly deployed in Google Colab or any other Tensorflow environment.
Why it matches plant phenotyping methods植物の気孔を顕微鏡画像から自動検出・個別境界推定し、形態解析に利用する画像ベース表現型解析手法を開発・検証しているため。
abstractIn this paper, we develop a fully automated, robust stomata detection algorithm which can also identify individual stomata boundaries
Reproduction assets foundThe paper's authors publicly release their complete Mask R-CNN stomata segmentation analysis code at the Smart-Robotic-Viticulture/MaskStomata GitHub repository, with explicit availability statements in multiple blocks. The microscope image datasets themselves are only available from the corresponding author on requestCode · publicThe complete code for the project can be accessed at: https://github.com/Smart-Robotic-Viticulture/MaskStomata .Open asset ↗Smart-Robotic-Viticulture/MaskStomatalines:136-157Code / dataset availability confirmedEurope PMC · checked 9 Sept 2026
The seeds of Arabidopsis thaliana become encapsulated by a layer of mucilage when imbibed. This polysaccharide-rich hydrogel is constituted of two layers, an outer layer that can be easily extracted with water and an inner layer that must be examined in situ in order to study its properties and structure in a non-destructive manner or disintegrated through hydrolysis or physical means in order to analyze its constituents. Mucilage production is an adaptive trait and we have exploited 19 natural accessions previously found to have atypical and varied outer mucilage characteristics. A detailed study using biochemical, histological and Time-Domain NMR analyses has been used to generate three related datasets covering 33 traits measured in four biological replicates. This data will be a rich resource for genetic, biochemical, structural and functional analyses investigating mucilage constituent polysaccharides or their role as adaptive traits.
Why it matches plant phenotyping methodsアラビドプシス種子の粘液形質を対象に、33形質・4反復の再利用可能なデータセットを生成した研究であり、植物形質データセットの構築が中心です。
abstractA detailed study using biochemical, histological and Time-Domain NMR analyses has been used to generate three related datasets covering 33 traits measured in four biological replicates.
Reproduction assets foundThe paper deposits its plant-phenotyping measurements in three Data INRAE datasets. Two of them (dataset 1: 33 mucilage/seed traits; dataset 3: individual microscopy measurements of mucilage and seed width) have DOIs matching allowed_urls entries and are directly citable public assets. Dataset 2's DOI (10.15454/EYABB2)Dataset · publicCambert, M. et al. Seed mucilage traits for Arabidopsis thaliana natural accessions with atypical outer mucilage - dataset 1. Portail
Data INRAE https://doi.org/10.15454/1MZ1ZC (2021).Open asset ↗10.15454/1MZ1ZCpdf-page:9 lines:1-70Dataset · publicBerger, A., Sallé, C. & North, H. M. Measurements of inner mucilage and seed width for Arabidopsis natural accessions - dataset 3.
Portail Data INRAE https://doi.org/10.15454/LBUN4X (2021).Open asset ↗Portail Data INRAE · 10.15454/LBUN4Xpdf-page:9 lines:1-70Code / dataset availability confirmedEurope PMC · Crossref · bioRxiv · checked 15 Sept 2026
MicroscopyCell / cellular structureTissueSegmentationGrowth / development / phenology
Background and Objective A variety of genetic mutations are known to affect cell proliferation and apoptosis during organism development, leading to structural birth defects such as facial clefting. Yet, the mechanisms how these alterations influence the development of the face remain unclear. Cell proliferation and its relation to shape variation can be studied in high detail using Light-Sheet Microscopy (LSM) imaging across a range of developmental time points. However, the large number of LSM images captured at cellular resolution precludes manual analysis. Thus, the aim of this work was to develop and evaluate automatic methods to segment tissues and proliferating cells in these images in an accurate and efficient way. Methods We developed, trained, and evaluated convolutional neural networks (CNNs) for segmenting tissues, cells, and specifically proliferating cells in LSM datasets. We compared the automatically extracted tissue and cell annotations to corresponding manual segmentations for three specific applications: (i) tissue segmentation (neural ectoderm and mesenchyme) in nuclear-stained LSM images, (ii) cell segmentation in nuclear-stained LSM images, and (iii) segmentation of proliferating cells in Phospho-Histone H3 (PHH3)-stained LSM images. Results The automatic CNN-based tissue segmentation method achieved a macro-average F-score of 0.84 compared to a macro-average F-score of 0.89 comparing corresponding manual segmentations from two observers. The automatic cell segmentation method in nuclear-stained LSM images achieved an F-score of 0.57, while comparing the manual segmentations resulted in an F-score of 0.39. Finally, the automatic segmentation method of proliferating cells in the PHH3-stained LSM datasets achieved an F-score of 0.56 for the automated method, while comparing the manual segmentations resulted in an F-score of 0.45. Conclusions The proposed automatic CNN-based framework for tissue and cell segmentation leads to results comparable to the inter-observer agreement, accelerating the LSM image analysis. The trained CNN models can also be applied for shape or morphological analysis of embryos, and more generally in other areas of cell biology.
Why it matches plant phenotyping methods発生中の胚の組織・細胞を対象に、ライトシート画像から形態関連の構造を自動抽出するCNN分割法を開発・評価しており、植物ではないため対象範囲外です。
abstractThus, the aim of this work was to develop and evaluate automatic methods to segment tissues and proliferating cells in these images in an accurate and efficient way.
Reproduction assets foundThe paper's authors explicitly state that their source code, software, and annotated LSM image datasets (DAPI-Tissue, DAPI-Cells, PHH3-Cells) are publicly available in their GitHub repositories, which directly reproduce this paper's segmentation models and analysis.Code · publicion. For segmentation of
proliferating cells, the U-net was trained using PHH3-stained images with
corresponding manual segmentations. Finally, the three segmentations are
combined to create maps of relative proliferation in the mesenchyme. The
source code, software, and annotated datasets have been made publicly avail-
able at https://github.com/lucaslovercio/LSMprocessing.2. Materials and Methods
2.1. Image acquisition
Five E9.5 and five E10.5 mice embryos were harvested and fixed overnight
in 4% paraformaldehyde. After fixation, they were processed for clearing
and staining. The clearing step followed the CUBIC protocol [23]. Briefly
described, embryos were incubated overnight in Cubic1/HOpen asset ↗lucaslovercio/LSMprocessing.2pdf-raw-page:5 lines:1-47Code · publicof proliferating cells, tissues, and total cells. One
CNN model was trained for each segmentation problem, and the quantita-
tive evaluation suggests that all three models lead to segmentation results
within the range of the inter-observer agreement. The source code, soft-
ware, and annotated datasets are publicly available at https://github.com/lucaslovercio/LSMprocessing. The methods developed in this work
are integral to the larger goal of improving the understanding of development
and morphogenesis and how perturbations to development result in diseases.
22
.
CC-BY-NC-ND 4.0 International license
available under a
(which was not certified by peer review) is the author/funder, who has graOpen asset ↗lucaslovercio/LSMprocessingpdf-raw-page:22 lines:1-45Code / dataset availability confirmedEurope PMC · checked 15 Sept 2026
The importance of fluorescence light microscopy for understanding cellular and sub-cellular structures and functions is undeniable. However, the resolution is limited by light diffraction (~200-250 nm laterally, ~500-700 nm axially). Meanwhile, super-resolution microscopy, such as structured illumination microscopy (SIM), is being applied more and more to overcome this restriction. Instead, super-resolution by stimulated emission depletion (STED) microscopy achieving a resolution of ~50 nm laterally and ~130 nm axially has not yet frequently been applied in plant cell research due to the required specific sample preparation and stable dye staining. Single-molecule localization microscopy (SMLM) including photoactivated localization microscopy (PALM) has not yet been widely used, although this nanoscopic technique allows even the detection of single molecules. In this study, we compared protein imaging within metaphase chromosomes of barley via conventional wide-field and confocal microscopy, and the sub-diffraction methods SIM, STED, and SMLM. The chromosomes were labeled by DAPI (4',6-diamidino-2-phenylindol), a DNA-specific dye, and with antibodies against topoisomerase IIα (Topo II), a protein important for correct chromatin condensation. Compared to the diffraction-limited methods, the combination of the three different super-resolution imaging techniques delivered tremendous additional insights into the plant chromosome architecture through the achieved increased resolution.
Why it matches plant phenotyping methods植物染色体の構造を対象に、複数の顕微鏡法を比較評価し、解像度向上による表現型（染色体アーキテクチャ）取得を中心課題としている。
abstractIn this study, we compared protein imaging within metaphase chromosomes of barley via conventional wide-field and confocal microscopy, and the sub-diffraction methods SIM, STED, and SMLM.
Reproduction assets foundThe paper's super-resolution microscopy measurements (SIM/STED/PALM imaging of barley metaphase chromosomes) are supported by supplementary materials, including PALM movies (Movies S1–S4) and supplementary figures, publicly available at the MDPI supplement URL. The main datasets themselves are only 'available from the Supplement · publicphotoactivated localization microscopy
rb
rabbit
RT
room temperature
SIM
structured illumination microscopy
SMLM
single molecule localization microscopy
STED
stimulated emission depletion
STORM
stochastic optical reconstruction microscopy
Topo II
topoisomerase IIα
Supplementary Materials
Supplementary Materials can be found at https://www.mdpi.com/1422-0067/22/4/1903/s1 .
Click here for additional data file.
Author Contributions
V.S. conceived the project. I.K., A.N., V.S. and K.W. conducted the study and processed the data. I.K., A.N. and V.S. wrote the manuscript. I.K., A.N., K.W., E.H. and V.S. discussed the results and contributed to manuscript writing. All authors have read and agreed tOpen asset ↗lines:237-298Code / dataset availability confirmedEurope PMC · checked 9 Sept 2026
Published1 Feb 2021Proceedings of the National Academy of Sciences of the United States of AmericaCited by 35 · OpenAlex ↗
Artificial mechanical perturbations affect chromatin in animal cells in culture. Whether this is also relevant to growing tissues in living organisms remains debated. In plants, aerial organ emergence occurs through localized outgrowth at the periphery of the shoot apical meristem, which also contains a stem cell niche. Interestingly, organ outgrowth has been proposed to generate compression in the saddle-shaped organ-meristem boundary domain. Yet whether such growth-induced mechanical stress affects chromatin in plant tissues is unknown. Here, by imaging the nuclear envelope in vivo over time and quantifying nucleus deformation, we demonstrate the presence of active nuclear compression in that domain. We developed a quantitative pipeline amenable to identifying a subset of very deformed nuclei deep in the boundary and in which nuclei become gradually narrower and more elongated as the cell contracts transversely. In this domain, we find that the number of chromocenters is reduced, as shown by chromatin staining and labeling, and that the expression of linker histone H1.3 is induced. As further evidence of the role of forces on chromatin changes, artificial compression with a MicroVice could induce the ectopic expression of H1.3 in the rest of the meristem. Furthermore, while the methylation status of chromatin was correlated with nucleus deformation at the meristem boundary, such correlation was lost in the h1.3 mutant. Altogether, we reveal that organogenesis in plants generates compression that is able to have global effects on chromatin in individual cells.
Why it matches plant phenotyping methods植物組織内の核変形を経時イメージングで定量化する解析パイプラインを開発し、核の形態状態を抽出しているため、表現型取得法が研究上実質的に中心である。
abstractHere, by imaging the nuclear envelope in vivo over time and quantifying nucleus deformation, we demonstrate the presence of active nuclear compression in that domain.
Reproduction assets foundThe paper's Data Availability statement deposits original confocal phenotyping data (meristem/nucleus imaging) in the Cambridge repository and provides the authors' segmentation/quantification analysis pipeline scripts on the Sainsbury Laboratory GitLab. Both are paper-specific, public, and actionable.Dataset · publicOriginal confocal data are available via the University of Cambridge Data Repository ( https://doi.org/10.17863/CAM.64310 ).Open asset ↗University of Cambridge Data Repository · 10.17863/CAM.64310lines:76-106Code · publicScripts for the analysis pipeline are available via the Sainsbury Laboratory GitLab repository ( https://gitlab.com/slcu/teamHJ/publications/fal_etal_2020 ).Open asset ↗Sainsbury Laboratory GitLab · slcu/teamHJ/publications/fal_etal_2020lines:76-106Code · publicScripts required to do the segmentation and quantitative analysis are provided via the Sainsbury Laboratory GitLab repository ( https://gitlab.com/slcu/teamhj/publications/fal_et_al_2021 ), where also a more detailed protocol for executing the steps of the pipeline is provided.Open asset ↗Sainsbury Laboratory GitLab · slcu/teamhj/publications/fal_et_al_2021lines:76-106Code / dataset availability confirmedCrossref · Europe PMC · checked 9 Sept 2026
Abstract Arbuscular mycorrhiza fungi (AMF) are beneficial soil fungi that can promote the growth of their host plants. Accurate quantification of AMF in plant roots is important because the level of colonization is often indicative of the activity of these fungi. Root colonization is traditionally measured with microscopy methods which visualize fungal structures inside roots. Microscopy methods are labor-intensive, and results depend on the observer. In this study, we present a relative qPCR method to quantify AMF in which we normalized the AMF qPCR signal relative to a plant gene. First, we validated the primer pair AMG1F and AM1 in silico, and we show that these primers cover most AMF species present in plant roots without amplifying host DNA. Next, we compared the relative qPCR method with traditional microscopy based on a greenhouse experiment with Petunia plants that ranged from very high to very low levels of AMF root colonization. Finally, by sequencing the qPCR amplicons with MiSeq, we experimentally confirmed that the primer pair excludes plant DNA while amplifying mostly AMF. Most importantly, our relative qPCR approach was capable of discriminating quantitative differences in AMF root colonization and it strongly correlated (Spearman Rho = 0.875) with quantifications by traditional microscopy. Finally, we provide a balanced discussion about the strengths and weaknesses of microscopy and qPCR methods. In conclusion, the tested approach of relative qPCR presents a reliable alternative method to quantify AMF root colonization that is less operator-dependent than traditional microscopy and offers scalability to high-throughput analyses.
Why it matches plant phenotyping methods植物根のAMF菌根 colonization を定量する相対qPCR法を開発・検証し、顕微鏡法との比較で性能を評価しているため、植物状態の取得手法が中心である。
abstractIn this study, we present a relative qPCR method to quantify AMF in which we normalized the AMF qPCR signal relative to a plant gene.
Reproduction assets foundThe paper provides two paper-specific public assets: the authors' analysis code repository on GitHub (R/DADA2/qPCR analysis workflow) and raw amplicon sequencing data deposited in the European Nucleotide Archive under study accession PRJEB20127 (sample SAMEA103939171), which contains the qPCR amplicon sequences used toCode · publicAll code is available under https://github.com/PMI-Basel/Bodenhausen_et_al_AMF_qPCR .Open asset ↗PMI-Basel/Bodenhausen_et_al_AMF_qPCRlines:90-98Code / dataset availability confirmedEurope PMC · Crossref · checked 15 Sept 2026
Tissue clearing methods combined with confocal microscopy have been widely used for studying developmental biology. In plants, ClearSee is a reliable clearing method that is applicable to a wide range of tissues and is suitable for gene expression analysis using fluorescent reporters, but its application to the Arabidopsis thaliana embryo, a model system to study morphogenesis and pattern formation, has not been described in the original literature. Here, we describe a ClearSee-based clearing protocol which is suitable for obtaining 3D images of Arabidopsis thaliana embryos. The method consists of embryo dissection, fixation, washing, clearing, and cell wall staining and enables high-quality 3D imaging of embryo morphology and expression of fluorescent reporters with the cellular resolution. Our protocol provides a reliable method that is applicable to the analysis of morphogenesis and gene expression patterns in Arabidopsis thaliana embryos.
Why it matches plant phenotyping methodsArabidopsis胚の形態を細胞解像度で3D取得するClearSeeベースのクリアリング・画像化プロトコルが研究の中心であり、植物表現型の取得方法を開発している。
abstractHere, we describe a ClearSee-based clearing protocol which is suitable for obtaining 3D images of Arabidopsis thaliana embryos.
Reproduction assets foundThe paper's supplementary materials include Movie S1, the Z-stack confocal image data (157 serial optical sections) used for the paper's 3D embryo visualization analysis, publicly available at the MDPI supplementary URL. No author analysis code or trained models are reported.Supplement · publicThe following are available online at https://www.mdpi.com/2223-7747/10/2/190/s1 , Movie S1: Z-stack images of 157 serial optical sections used for Figure 2 ; Table S1: Primers used in this study.
Click here for additional data file.
Author Contributions
Conceptualization, M.A.; methodology, M.A.; validation, A.I. and M.A.; formal analysis, A.I.; investigation, A.I., M.Y., T.S., A.O., and M.A.; resources, TOpen asset ↗lines:58-85Code / dataset availability confirmedEurope PMC · checked 9 Sept 2026
Abstract Background: Stomata analysis using microscope imagery provides important insight into plant physiology, health and the surrounding environmental conditions. Plant scientists are now able to conduct automated high-throughput analysis of stomata in microscope data, however, existing detection methods are sensitive to the appearance of stomata in the training images, thereby limiting general applicability. In addition, existing methods only generate bounding-boxes around detected stomata, which require users to implement additional image processing steps to study stomata morphology. In this paper, we develop a fully automated, robust stomata detection algorithm which can also identify individual stomata boundaries regardless of the plant species, sample collection method, imaging technique and magnification level. Results: The proposed solution consists of three stages. First, the input image is pre-processed to remove any colour space biases occurring from different sample collection and imaging techniques. Then, a Mask R-CNN is applied to estimate individual stomata boundaries. The feature pyramid network embedded in the Mask R-CNN is utilised to identify stomata at different scales. Finally, a statistical filter is implemented at the Mask R-CNN output to reduce the number of false positive generated by the network. The algorithm was tested using 16 datasets from 12 sources, containing over 60,000 stomata. For the first time in this domain, the proposed solution was tested against 7 microscope datasets never seen by the algorithm to show the generalisability of the solution. Results indicated that the proposed approach can detect stomata with a precision, recall, and F-score of 95.10\%, 83.34\%, and 88.61\%, respectively. A separate test conducted by comparing estimated stomata boundary values with manually measured data showed that the proposed method has an IoU score of 0.70; a 7\% improvement over the bounding-box approach. Conclusions: The proposed method shows robust performance across multiple microscope image datasets of different quality and scale. This generalised stomata detection algorithm allows plant scientists to conduct stomata analysis whilst eliminating the need to re-label and re-train for each new dataset. The open-source code shared with this project can be directly deployed in Google Colab or any other Tensorflow environment.
Why it matches plant phenotyping methods気孔の画像から個体境界を自動抽出する汎用的な画像解析手法を開発し、多数のデータセットで汎化性能と手動測定との一致を検証しており、植物表現型取得が中心である。
abstractIn this paper, we develop a fully automated, robust stomata detection algorithm which can also identify individual stomata boundaries regardless of the plant species, sample collection method, imaging technique and magnification level.
Reproduction assets foundThe paper's complete analysis code (Mask R-CNN stomata segmentation pipeline, training setup, fine-tuning instructions) is publicly available on the authors' GitHub repository. The microscope image datasets themselves are only available on request from the corresponding author, so they do not qualify as public assets.Code · publicThe complete code for the project can be accessed at: :
https://github.com/Smart-Robotic-Viticulture/MaskStomata.Open asset ↗Smart-Robotic-Viticulture/MaskStomatapdf-page:19 lines:1-68Code / dataset availability confirmedEurope PMC · Crossref · bioRxiv · checked 15 Sept 2026
Capturing complete internal anatomies of plant organs and tissues within their relevant morphological context remains a key challenge in plant science. While plant growth and development are inherently multiscale, conventional light, fluorescence, and electron microscopy platforms are typically limited to imaging of plant microstructure from small flat samples that lack direct spatial context to, and represent only a small portion of, the relevant plant macrostructures. We demonstrate technical advances with a lab-based X-ray microscope (XRM) that bridge the imaging gap by providing multiscale high-resolution 3D volumes of intact plant samples from the cell to whole plant level. Serial imaging of a single sample is shown to provide sub-micron 3D volumes co-registered with lower magnification scans for explicit contextual reference. High quality 3D volume data from our enhanced methods facilitate more sophisticated and effective computational segmentation and analyses than have previously been employed for X-ray based imaging. Advances in sample preparation make multimodal correlative imaging workflows possible, where a single resin-embedded plant sample is scanned via XRM to generate a 3D cell-level map, and then used to identify and zoom in on sub-cellular regions of interest for high resolution scanning electron microscopy. In total, we present the methodologies for use of XRM in the multiscale and multimodal analysis of 3D plant features using numerous economically and scientifically important plant systems.
Why it matches plant phenotyping methods植物試料の細胞から個体までを対象に、X線顕微鏡によるマルチスケール3D画像取得、試料調製、計算セグメンテーション、相関イメージングの方法論を中心に提示しており、植物形態の取得・解析法が明確に中心です。
abstractWe demonstrate technical advances with a lab-based X-ray microscope (XRM) that bridge the imaging gap by providing multiscale high-resolution 3D volumes of intact plant samples from the cell to whole plant level.
Reproduction assets foundThe preprint points to a public figshare collection containing the paper's high-resolution XRM image stacks ('flythroughs') and videos of the 3D plant datasets, which directly reproduce the paper's phenotyping imaging measurements. No author analysis code or trained model checkpoint is explicitly deposited; the deep-seDataset · publicof these improved techniques will
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make a significant contribution to plant biology, expanding the reach of XRM as a
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routine tool for 3D imaging for plant scientists.
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RESULTS1
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Meristem Biology
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high-resolution image stacks (“flythroughs”) and videos portraying the 3D data sets can be found here:
https://figshare.com/s/944efc8832e47fd4f203
.
CC-BY-NC 4.0 International license
available under a
(which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made
The copyright holder for this preprint
this version posted December 22, 2020.
;
https://doi.org/10.1101/2020.12.18.423480
doiOpen asset ↗figsharepdf-raw-page:4 lines:1-64Code / dataset availability confirmedEurope PMC · checked 9 Sept 2026
NucleusJ 1.0, an ImageJ plugin, is a useful tool to analyze nuclear morphology and chromatin organization in plant and animal cells. NucleusJ 2.0 is a new release of NucleusJ, in which image processing is achieved more quickly using a command-lineuser interface. Starting with large collection of 3D nuclei, segmentation can be performed by the previously developed Otsu-modified method or by a new 3D gift-wrapping method, taking better account of nuclear indentations and unstained nucleoli. These two complementary methods are compared for their accuracy by using three types of datasets available to the community at https://www.brookes.ac.uk/indepth/images/ . Finally, NucleusJ 2.0 was evaluated using original plant genetic material by assessing its efficiency on nuclei stained with DNA dyes or after 3D-DNA Fluorescence in situ hybridization. With these improvements, NucleusJ 2.0 permits the generation of large user-curated datasets that will be useful for software benchmarking or to train convolution neural networks.
Why it matches plant phenotyping methods植物細胞核の3D形態・クロマチン構造を画像から抽出するソフトウェアの開発、セグメンテーション手法の比較検証、植物試料での評価が中心であり、植物フェノタイピング手法に該当する。
abstractNucleusJ 2.0 is a new release of NucleusJ, in which image processing is achieved more quickly using a command-lineuser interface.
Reproduction assets foundThe paper's 3D nuclear bio-imaging datasets (7,313 images across six dataset types: digitized spheres, fluorescent microspheres, nuclear morphology, chromatin organization, and DNA FISH) are stored in the public OMERO-FSU repository under accession IDP3006_Dubos-Desset_2020, accessible via the INDEPTH website. The NuceDataset · publicDatasets were stored at OMERO-Florida State University (OMERO-FSU), a public repository under the accession number IDP3006 Dubos–Desset Nucleus 2020 that can be accessed through the INDEPTH COST-Action (CA16212) website at https://www.brookes.ac.uk/indepth/images/ . The INDEPTH image webpage provides a guideline to access and download the datasets that are freely available for research purposes.Open asset ↗OMERO-FSUlines:180-188Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
Published1 Dec 2020Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of CanadaCited by 16 · OpenAlex ↗
Phytoliths can be an important source of information related to environmental and climatic change, as well as to ancient plant use by humans, particularly within the disciplines of paleoecology and archaeology. Currently, phytolith identification and categorization is performed manually by researchers, a time-consuming task liable to misclassifications. The automated classification of phytoliths would allow the standardization of identification processes, avoiding possible biases related to the classification capability of researchers. This paper presents a comparative analysis of six classification methods, using digitized microscopic images to examine the efficacy of different quantitative approaches for characterizing phytoliths. A comprehensive experiment performed on images of 429 phytoliths demonstrated that the automatic phytolith classification is a promising area of research that will help researchers to invest time more efficiently and improve their recognition accuracy rate.
Why it matches plant phenotyping methods植物由来の植物珪酸体を対象に、顕微鏡画像からの自動識別・分類手法を比較評価しており、画像ベースの形態的特徴抽出が研究の中心です。
abstractThis paper presents a comparative analysis of six classification methods, using digitized microscopic images to examine the efficacy of different quantitative approaches for characterizing phytoliths.
Reproduction assets foundThe paper's phytolith photomicrograph dataset (429 images across 8 morphotypes) is explicitly stated to be publicly available at the UPF repository, and the authors' analysis code is shared on GitHub with explicit availability language. Both are paper-specific, public, and actionable.Dataset · publiccaptured from the side view.
The total number of photomicrographs obtained for each mor-
photype is shown in Table 1. Only nonarticulated (not attached to
any other phytoliths) were considered and just one photomicro-
graph per phytolith was recorded. The total number of samples
was 429. All the images are publicly available at https://reposi-tori.upf.edu/handle/10230/44939, all the morphotypes have at
least 50 samples, and the dataset is fairly balanced (i.e., there is
a similar number of samples per class).
The image of each phytolith was digitized, using an open-
source web annotation tool called VGG Image Annotator.1
This
tool allows a researcher to create a control-points based contour
Open asset ↗reposi-tori.upf.edu · 10230/44939pdf-raw-page:3 lines:1-92Code · publicclassification process. Even though several researchers have
attempted to create automatic tools for the identification of
archaeobotanical remains, none of the attempts has produced a
tool that is accessible online or as a downloadable app. We are
sharing the code used in our research (which is accessible at
https://github.com/alvarag/AutomaticPhytolithClassification) to
stimulate other researchers to join in the effort to build a real
and functional tool that can be trained online, increasing its
accuracy.
Future Research Lines
The development of new features and the application of feature
selection techniques are some of the research avenues we are plan-
ning to explore. It would be imporOpen asset ↗github.com/alvarag/AutomaticPhytolithClassificationpdf-raw-page:9 lines:1-85Code / dataset availability confirmedEurope PMC · checked 9 Sept 2026
The CO 2 and water vapor exchange between leaf and atmosphere are relevant for plant physiology. This process is done through the stomata. These structures are fundamental in the study of plants since their properties are linked to the evolutionary process of the plant, as well as its environmental and phytohormonal conditions. Stomatal detection is a complex task due to the noise and morphology of the microscopic images. Although in recent years segmentation algorithms have been developed that automate this process, they all use techniques that explore chromatic characteristics. This research explores a unique feature in plants, which corresponds to the stomatal spatial distribution within the leaf structure. Unlike segmentation techniques based on deep learning tools, we emphasize the search for an optimal threshold level, so that a high percentage of stomata can be detected, independent of the size and shape of the stomata. This last feature has not been reported in the literature, except for those results of geometric structure formation in the salt formation and other biological formations.
Why it matches plant phenotyping methods葉の顕微鏡画像から気孔を自動セグメンテーションする手法の開発が中心であり、植物の形態的形質取得に直接関わる。
titleAutomatic Stomatal Segmentation Based on Delaunay-Rayleigh Frequency Distance.
Reproduction assets foundThe paper's Supplementary Materials section explicitly states that the DRTB solution (authors' analysis code) is available online at https://github.com/mlacarrasco/drtb and that the images database (stomatal microscopy images used for phenotyping) is available at https://github.com/mlacarrasco/drtb/tree/main/database. Dataset · publicOur solution can be accessed online at https://github.com/mlacarrasco/drtb , and images database are available online at https://github.com/mlacarrasco/drtb/tree/main/database .Open asset ↗mlacarrasco/drtblines:61-134Code / dataset availability confirmedCrossref · Europe PMC · checked 9 Sept 2026
Abstract We present a new large-scale three-fold annotated microscopy image dataset, aiming to advance the plant cell biology research by exploring different cell microstructures including cell size and shape, cell wall thickness, intercellular space, etc. in deep learning (DL) framework. This dataset includes 9,811 unstained and 6,127 stained (safranin-o, toluidine blue-o, and lugol’s-iodine) images with three-fold annotation including physical, morphological, and tissue grading based on weight, different section area, and tissue zone respectively. In addition, we prepared ground truth segmentation labels for three different tuber weights. We have validated the pertinence of annotations by performing multi-label cell classification, employing convolutional neural network (CNN), VGG16, for unstained and stained images. The accuracy has been achieved up to 0.94, while, F2-score reaches to 0.92. Furthermore, the ground truth labels have been verified by semantic segmentation algorithm using UNet architecture which presents the mean intersection of union up to 0.70. Hence, the overall results show that the data are very much efficient and could enrich the domain of microscopy plant cell analysis for DL-framework.
Why it matches plant phenotyping methodsジャガイモ塊茎の細胞形態・組織特性を対象とする大規模画像データセットを構築し、分類・セグメンテーションで検証しており、植物フェノタイピング用データ資源が中心である。
titleA large-scale optical microscopy image dataset of potato tuber for deep learning based plant cell assessment
Reproduction assets foundThe paper's potato tuber microscopy image dataset (raw stained/unstained images plus ground truth segmentation labels) is publicly deposited on figshare by the authors.Dataset · publicThis dataset is publicly available on figshare47 (https://doi.org/10.6084/m9.figshare.c.4955669) which can be
downloaded as a zip file.Open asset ↗figshare · 10.6084/m9.figshare.c.4955669pdf-page:5 lines:1-35Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
The plant cell wall (PCW) is a pecto-cellulosic extracellular matrix that envelopes the plant cell. By integrating extra-and intra-cellular cues, PCW mediates a plethora of essential physiological functions. Notably, it permits controlled and oriented tissue growth by tuning its local mechano-chemical properties. To refine our knowledge of these essential properties of PCW, we need an appropriate tool for the accurate observation of the native ( in muro ) structure of the cell wall components. The label-free techniques, such as AFM, EM, FTIR, and Raman microscopy, are used; however, they either do not have the chemical or spatial resolution. Immunolabeling with electron microscopy allows observation of the cell wall nanostructure, however, it is mostly limited to single and, less frequently, multiple labeling. Immunohistochemistry (IHC) is a versatile tool to analyze the distribution and localization of multiple biomolecules in the tissue. The subcellular resolution of chemical changes in the cell wall component can be observed with standard diffraction-limited optical microscopy. Furthermore, novel chemical imaging tools such as multicolor 3D dSTORM (Three-dimensional, direct Stochastic Optical Reconstruction Microscopy) nanoscopy makes it possible to resolve the native structure of the cell wall polymers with nanometer precision and in three dimensions. Here we present a protocol for preparing multi-target immunostaining of the PCW components taking as example Arabidopsis thaliana , Star fruit ( Averrhoa carambola) , and Maize thin tissue sections. This protocol is compatible with the standard confocal microscope, dSTORM nanoscope, and can also be implemented for other optical nanoscopy such as STED (Stimulated Emission Depletion Microscopy). The protocol can be adapted for any other subcellular compartments, plasma membrane, cytoplasmic, and intracellular organelles.
Why it matches plant phenotyping methods植物細胞壁成分の多重免疫染色と共焦点・超解像イメージングのプロトコル開発が中心であり、細胞壁構造・局在という植物状態の取得法を扱う。
abstractHere we present a protocol for preparing multi-target immunostaining of the PCW components
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Code · publicGrafeo (Custom made software for dSTORM data analysis and visualization, https://github.com/inatamara/Grafeo-dSTORM-analysis- (Open asset ↗inatamara/Grafeo-dSTORM-analysis-lines:208-269Code / dataset availability confirmedEurope PMC · checked 9 Sept 2026
Arbuscular mycorrhizal fungi function as conduits for underground nutrient transport. While the fungal partner is dependent on the plant host for its carbon (C) needs, the amount of nutrients that the fungus allocates to hosts can vary with context. Because fungal allocation patterns to hosts can change over time, they have historically been difficult to quantify accurately. We developed a technique to tag rock phosphorus (P) apatite with fluorescent quantum-dot (QD) nanoparticles of three different colors, allowing us to study nutrient transfer in an in vitro fungal network formed between two host roots of different ages and different P demands over a 3-week period. Using confocal microscopy and raster image correlation spectroscopy, we could distinguish between P transfer from the hyphae to the roots and P retention in the hyphae. By tracking QD-apatite from its point of origin, we found that the P demands of the younger root influenced both: (1) how the fungus distributed nutrients among different root hosts and (2) the storage patterns in the fungus itself. Our work highlights that fungal trade strategies are highly dynamic over time to local conditions, and stresses the need for precise measurements of symbiotic nutrient transfer across both space and time.
Why it matches plant phenotyping methods量子ドット標識と共焦点画像解析を開発し、植物根へのリン移行および菌根内保持を時空間的に定量する手法が研究の中心であるため、植物の栄養生理状態を測定するフェノタイピング手法として含める。
abstractWe developed a technique to tag rock phosphorus (P) apatite with fluorescent quantum-dot (QD) nanoparticles of three different colors
Reproduction assets foundThe paper's authors publicly deposited all data, scripts, and analysis for this study in a GitHub repository, explicitly stated in the Methods. This is a paper-specific, publicly actionable code/data asset reproducing the paper's QD-apatite phenotyping measurements and statistical analysis.Code · publicWe performed all statistical analysis in R version 3.6.1 [ 48 ]. All data, scripts, and analysis are available at: https://github.com/anoukvantpadje/Two_roots .Open asset ↗anoukvantpadje/Two_rootslines:57-192Code / dataset availability confirmedCrossref · checked 9 Sept 2026
Stomata are microscopic pores on the plant epidermis that regulate the water content and CO2 levels in leaves. Thus, they play an important role in plant growth and development. Currently, most of the common methods for the measurement of pore anatomy parameters involve manual measurement or semi-automatic analysis technology, which makes it difficult to achieve high-throughput and automated processing. This paper presents a method for the automatic segmentation and parameter calculation of stomatal pores in microscope images of plant leaves based on deep convolutional neural networks. The proposed method uses a type of convolutional neural network model (Mask R-CNN (region-based convolutional neural network)) to obtain the contour coordinates of the pore regions in microscope images of leaves. The anatomy parameters of pores are then obtained by ellipse fitting technology, and the quantitative analysis of pore parameters is implemented. Stomatal microscope image datasets for black poplar leaves were obtained using a large depth-of-field microscope observation system, the VHX-2000, from Keyence Corporation. The images used in the training, validation, and test sets were taken randomly from the datasets (562, 188, and 188 images, respectively). After 10-fold cross validation, the 188 test images were found to contain an average of 2278 pores (pore widths smaller than 0.34 μm (1.65 pixels) were considered to be closed stomata), and an average of 2201 pores were detected by our network with a detection accuracy of 96.6%, and the intersection of union (IoU) of the pores was 0.82. The segmentation results of 2201 stomatal pores of black poplar leaves showed that the average measurement accuracies of the (a) pore length, (b) pore width, (c) area, (d) eccentricity, and (e) degree of stomatal opening, with a ratio of width-to-maximum length of a stomatal pore, were (a) 94.66%, (b) 93.54%, (c) 90.73%, (d) 99.09%, and (e) 92.95%, respectively. The proposed stomatal pore detection and measurement method based on the Mask R-CNN can automatically measure the anatomy parameters of pores in plants, thus helping researchers to obtain accurate stomatal pore information for leaves in an efficient and simple way.
Why it matches plant phenotyping methods葉の気孔画像から形態・開口状態を自動抽出・定量する画像解析手法を開発し、精度検証しており、植物フェノタイピング手法が研究の中心です。
abstractThis paper presents a method for the automatic segmentation and parameter calculation of stomatal pores in microscope images of plant leaves based on deep convolutional neural networks.
Reproduction assets foundThe paper's authors explicitly state that the complete project code for the Mask R-CNN-based stomatal pore detection and measurement method is publicly available on GitHub. The image datasets themselves are not stated as deposited by the authors (the generalization datasets are cited prior work, Stomatacounter [39]).Code · publicThe complete code for the project can be accessed at
https://github.com/lijunyu159/stomatal_pore_measurement-MaskRCNN (accessed on 15 July 2020).Open asset ↗lijunyu159/stomatal_pore_measurement-MaskRCNNpdf-page:9 lines:1-57Code / dataset availability confirmedOpenAlex · Europe PMC · checked 15 Sept 2026
Understanding plant growth processes is important for many aspects of biology and food security. Automating the observations of plant development-a process referred to as plant phenotyping-is increasingly important in the plant sciences, and is often a bottleneck. Automated tools are required to analyze the data in microscopy images depicting plant growth, either locating or counting regions of cellular features in images. In this paper, we present to the plant community an introduction to and exploration of two machine learning approaches to address the problem of marker localization in confocal microscopy. First, a comparative study is conducted on the classification accuracy of common conventional machine learning algorithms, as a means to highlight challenges with these methods. Second, a 3D (volumetric) deep learning approach is developed and presented, including consideration of appropriate loss functions and training data. A qualitative and quantitative analysis of all the results produced is performed. Evaluation of all approaches is performed on an unseen time-series sequence comprising several individual 3D volumes, capturing plant growth. The comparative analysis shows that the deep learning approach produces more accurate and robust results than traditional machine learning. To accompany the paper, we are releasing the 4D point annotation tool used to generate the annotations, in the form of a plugin for the popular ImageJ (FIJI) software. Network models and example datasets will also be available online.
Why it matches plant phenotyping methods植物の共焦点画像から細胞周期マーカーを自動検出・分割する機械学習手法を開発し、定量評価・比較検証しているため、植物表現型取得が中心である。
abstractAutomating the observations of plant development-a process referred to as plant phenotyping-is increasingly important in the plant sciences
Reproduction assets foundThe paper's Data Availability Statement explicitly deposits the confocal image datasets, annotations, and the annotation plugin in a public GitLab repository, matching the allowed URL.Code · publicample. As a pixel-wise segmentation is already produced by the network, but refined in post-processing to a single location in space, the network is already partially capable of generating meaningful 3D shape labels.
Data Availability Statement
The datasets and plugin used for this study can be found in the GitLab repository at https://gitlab.com/faraz.khan1/volumetric-segmentation-of-confocal-images .
Author Contributions
FK designed and implemented the computational algorithms, models and experiments. MP wrote the annotation tool and provided guidance. UV performed biological experiments and annotation. AF managed the project and helped design the approaches, with MP and FK. All authors coOpen asset ↗gitlab.com/faraz.khan1/volumetric-segmentation-of-confocal-imageslines:320-348Code / dataset availability confirmedEurope PMC · checked 14 Sept 2026
Plant leaf stomata are the gatekeepers of the atmosphere-plant interface and are essential building blocks of land surface models as they control transpiration and photosynthesis. Although more stomatal trait data are needed to significantly reduce the error in these model predictions, recording these traits is time-consuming, and no standardized protocol is currently available. Some attempts were made to automate stomatal detection from photomicrographs; however, these approaches have the disadvantage of using classic image processing or targeting a narrow taxonomic entity which makes these technologies less robust and generalizable to other plant species. We propose an easy-to-use and adaptable workflow from leaf to label. A methodology for automatic stomata detection was developed using deep neural networks according to the state of the art and its applicability demonstrated across the phylogeny of the angiosperms.We used a patch-based approach for training/tuning three different deep learning architectures. For training, we used 431 micrographs taken from leaf prints made according to the nail polish method from herbarium specimens of 19 species. The best-performing architecture was tested on 595 images of 16 additional species spread across the angiosperm phylogeny.The nail polish method was successfully applied in 78% of the species sampled here. The VGG19 architecture slightly outperformed the basic shallow and deep architectures, with a confidence threshold equal to 0.7 resulting in an optimal trade-off between precision and recall. Applying this threshold, the VGG19 architecture obtained an average F -score of 0.87, 0.89, and 0.67 on the training, validation, and unseen test set, respectively. The average accuracy was very high (94%) for computed stomatal counts on unseen images of species used for training.The leaf-to-label pipeline is an easy-to-use workflow for researchers of different areas of expertise interested in detecting stomata more efficiently. The described methodology was based on multiple species and well-established methods so that it can serve as a reference for future work.
Why it matches plant phenotyping methods葉の顕微画像から気孔を自動検出・計数する画像解析ワークフローを開発し、多様な植物種で性能検証しており、植物フェノタイピング手法が研究の中心です。
abstractA methodology for automatic stomata detection was developed using deep neural networks
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産2件を確認しました。Dataset · publicAll light microscope images used in this study are made freely accessible on Zenodo under the CC‐by license ( http://doi.org/10.5281/zenodo.3579227 ).Open asset ↗Zenodo · 10.5281/zenodo.3579227lines:186-229Dataset · publicThe example image set can also be downloaded here: http://doi.org/10.5281/zenodo.3902280 .Open asset ↗Zenodo · 10.5281/zenodo.3902280lines:186-229Code / dataset availability confirmedCrossref · checked 14 Sept 2026
Lignocellulosic biomass (LB) is recalcitrant to enzymatic hydrolysis due to its compact and complex cell wall structure. To identify the parameters behind LB recalcitrance, experimental data over hydrolysis time must be collected. Here, we describe a novel method to collect time-lapse images during cell wall deconstruction by enzymatic hydrolysis. The protocol includes instructions for sample preparation, layout of a custom designed incubation chamber and instructions for confocal time lapse acquisition. The protocol sets out a detailed plan where cross-sections of untreated and pretreated poplar samples are mounted in a sealed frame containing a buffer and an enzymatic cocktail. The sealed frame is then placed into an incubator to maintain the sample at a constant temperature of 50 °C, which is optimal for enzymatic reaction while avoiding enzymatic cocktail evaporation. Using lignin natural autofluorescence, confocal z-stacks of untreated and pretreated samples were acquired at regular time intervals during enzymatic hydrolysis for 24 h. Acquisition parameters were optimized to compromise between image resolution and reduced photo-bleaching. The acquired image might then be processed by further development of algorithms to extract precise quantitative information on cell wall deconstruction. This protocol is an important first step towards elucidating the underlying parameters of LB recalcitrance by allowing the acquisition of high-quality images of LB hydrolysis for extracting quantitative data on LB deconstruction.
Why it matches plant phenotyping methodsポプラ細胞壁の分解状態を時系列の共焦点3D画像で取得するプロトコル自体が中心であり、植物組織状態の定量的表現型抽出を可能にするため。
abstractHere, we describe a novel method to collect time-lapse images during cell wall deconstruction by enzymatic hydrolysis.
Reproduction assets foundThe paper's authors state that the scripts for computing photobleaching signal loss and image registration/analysis are publicly available in the FARE Laboratory GitLab repository, with an explicit URL matching an allowed URL.Code · publicels’ intensity
reduction in confocal image) between successive z-stacks. The signal loss was computed by subtracting
the voxels’ intensities between the registered floating image, It ◦ T It ←It+∆t , and the reference image It+∆t
and summing up the subtracted values (Scripts are available at the FARE Laboratory Gitlab Repository
https://gitlab.com/farelab/teamyr/publications/zoghlami_et_al_sus_chem_2020).
4. Results
Using the protocol, we acquired confocal images of pretreated poplar samples during hydrolysis
(Figure 7). We could visually observe that the cell walls gradually degraded over time. To illustrate
the advantages offered by using this protocol to achieve a quantitative characterizaOpen asset ↗gitlab.com/farelab/teamyr/publications/zoghlami_et_al_sus_chem_2020pdf-layout-page:8 lines:1-42Code / dataset availability confirmedCrossref · Europe PMC · checked 9 Sept 2026
Quantitative analysis of plant and animal morphogenesis requires accurate segmentation of individual cells in volumetric images of growing organs. In the last years, deep learning has provided robust automated algorithms that approach human performance, with applications to bio-image analysis now starting to emerge. Here, we present PlantSeg, a pipeline for volumetric segmentation of plant tissues into cells. PlantSeg employs a convolutional neural network to predict cell boundaries and graph partitioning to segment cells based on the neural network predictions. PlantSeg was trained on fixed and live plant organs imaged with confocal and light sheet microscopes. PlantSeg delivers accurate results and generalizes well across different tissues, scales, acquisition settings even on non plant samples. We present results of PlantSeg applications in diverse developmental contexts. PlantSeg is free and open-source, with both a command line and a user-friendly graphical interface.
Why it matches plant phenotyping methods植物組織を細胞単位で抽出する画像解析パイプラインを開発し、異なる組織・スケール・撮像条件で性能を示しているため、植物フェノタイピング手法が中心である。
abstractHere, we present PlantSeg, a pipeline for volumetric segmentation of plant tissues into cells.
Reproduction assets foundThe paper publicly deposits all plant phenotyping image/ground-truth datasets on OSF (https://osf.io/uzq3w), including ovule, lateral root, meristem, and leaf confocal/light-sheet volumes with hand-curated segmentations, and releases the PlantSeg analysis code and pre-trained 3D U-Net models on GitHub.Dataset · publicAll datasets used to support the findings of this study have been deposited in https://osf.io/uzq3w .Open asset ↗osf.io/uzq3wlines:38-47Code · publicThe code used for training and inference can be found at Wolny, 2020b
https://github.com/wolny/pytorch-3dunet copy archived at https://github.com/elifesciences-publications/pytorch-3dunet .Open asset ↗GitHub · wolny/pytorch-3dunetlines:212-223Code / dataset availability confirmedEurope PMC · checked 9 Sept 2026
Optical Projection Tomography (OPT) is a powerful three-dimensional imaging technique used for the observation of millimeter-scaled biological samples, compatible with bright-field and fluorescence contrast. OPT is affected by spatially variant artifacts caused by the fact that light diffraction is not taken into account by the straight-light propagation models used for reconstruction. These artifacts hinder high-resolution imaging with OPT. In this work we show that, by using a multiview imaging approach, a 3D reconstruction of the bright-field contrast can be obtained without the diffraction artifacts typical of OPT, drastically reducing the amount of acquired data, compared to previously reported approaches. The method, purely based on bright-field contrast of the unstained sample, provides a comprehensive picture of the sample anatomy, as demonstrated in vivo on Arabidopsis thaliana and zebrafish embryos. Furthermore, this bright-field reconstruction can be implemented on practically any multi-view light-sheet fluorescence microscope without complex hardware modifications or calibrations, complementing the fluorescence information with tissue anatomy.
Why it matches plant phenotyping methods多視点取得と画像融合による3D明視野再構成法を開発し、Arabidopsisの解剖学的形態を実証しているため、植物形態の取得手法が中心です。
abstractIn this work we show that, by using a multiview imaging approach, a 3D reconstruction of the bright-field contrast can be obtained without the diffraction artifacts typical of OPT
Reproduction assets foundThe paper's authors state that the Python sample code implementing their bright-field multi-view reconstruction (used for the Arabidopsis thaliana and zebrafish phenotyping/imaging analysis) is publicly available on GitHub under the authors' account. The GitHub URL in the text contains formatting artifacts and does notCode · publicData processing was performed in Python; a sample code is available on GitHubOpen asset ↗pdf-page:7 lines:1-55Code / dataset availability confirmedEurope PMC · checked 9 Sept 2026
Plasmodesmata are small channels that connect plant cells. While recent technological advances have facilitated analysis of the ultrastructure of these channels, there are limitations to efficiently addressing their presence over an entire cellular interface. Here, we highlight the value of serial block electron microscopy for this purpose. We developed a computational pipeline to study plasmodesmata distributions and detect the presence/absence of plasmodesmata clusters, or pit fields, at the phloem unloading interfaces of Arabidopsis ( Arabidopsis thaliana ) roots. Pit fields were visualized and quantified. As the wall environment of plasmodesmata is highly specialized, we also designed a tool to extract the thickness of the extracellular matrix at and outside of plasmodesmata positions. We detected and quantified clear wall thinning around plasmodesmata with differences between genotypes, including the recently published plm-2 sphingolipid mutant. Our tools open avenues for quantitative approaches in the analysis of symplastic trafficking.
Why it matches plant phenotyping methods植物組織の電子顕微鏡画像から原形質連絡の分布、クラスター、細胞壁厚を定量化する計算パイプラインとツールを開発しており、植物形態・構造形質の取得が中心です。
abstractWe developed a computational pipeline to study plasmodesmata distributions and detect the presence/absence of plasmodesmata clusters, or pit fields
Reproduction assets foundThe paper's authors publicly released their Matlab plugins for plasmodesmata distribution and cell-wall thickness analysis on GitHub, a guided R analysis pipeline tutorial, and the Col-0 SB-EM data sets with segmented wall models and PD annotations on Figshare. Generic tools (MIB, matGeom, CRAN packages) and the EMPIARCode · publicA guided tutorial with all the necessary code for this analysis is available at https://andreapaterlini.github.io/Plasmodesmata_dist_wall/ (last accessed March 2020).Open asset ↗lines:148-159Dataset · publicThe Col-0 data sets used in this article, with corresponding models and annotations, are available on Figshare ( https://doi.org/10.6084/m9.figshare.12488702.v1 ). They can be used as example data sets to test our pipeline.Open asset ↗figshare · 10.6084/m9.figshare.12488702.v1lines:148-159Code / dataset availability confirmedbioRxiv · checked 15 Sept 2026
Recent years have witnessed a convergence of data and methods that allow us to approximate the shape, size, and functional attributes of biological organisms. This is not only limited to traditional model species: given the ability to culture and visualize a specific organism, we can capture both its structural and functional attributes. We present a quantitative model for the colonial diatom Bacillaria paradoxa, an organism that presents a number of unique attributes in terms of form and function. To acquire a digital model of B. paradoxa, we extract a series of quantitative parameters from microscopy videos from both primary and secondary sources. These data are then analyzed using a variety of techniques, including two rival deep learning approaches. We provide an overview of neural networks for non-specialists as well as present a series of analysis on Bacillaria phenotype data. The application of deep learning networks allows for two analytical purposes. Application of the DeepLabv3 pre-trained model extracts phenotypic parameters describing the shape of cells constituting Bacillaria colonies. Application of a semantic model trained on nematode embryogenesis data (OpenDevoCell) provides a means to analyze masked images of potential intracellular features. We also advance the analysis of Bacillaria colony movement dynamics by using templating techniques and biomechanical analysis to better understand the movement of individual cells relative to an entire colony. The broader implications of these results are presented, with an eye towards future applications to both hypothesis-driven studies and theoretical advancements in understanding the dynamic morphology of Bacillaria.
Why it matches plant phenotyping methods珪藻の顕微鏡動画から形態・細胞内特徴・群体運動を抽出する画像処理および深層学習手法が研究の中心であり、植物表現型解析手法の開発に該当する。
abstractTo acquire a digital model of B. paradoxa, we extract a series of quantitative parameters from microscopy videos from both primary and secondary sources.
Reproduction assets foundThe paper's Bacillaria phenotyping data and analysis assets are publicly available: raw data, processed numeric/image data, and code in the authors' Digital-Bacillaria GitHub repository; skeleton-creation scripts in the Image-Skeletons subrepository; the OpenDevoCell segmentation platform (GitHub and web app); and a GfDataset · publicstrains contained in our primary data (videos) have been harvested from the Neckar
river in Germany (49°04'41.8"N 9°09'17.9"E). Samples were collected on September 14, 2019.
The average size of each cell (filament) is approximately 81µm.
Data Availability
Select unprocessed (raw) data are available at our Github repository
(https://github.com/devoworm/Digital-Bacillaria), processed numeric and image data (numeric
tables and skeletonized images), and select video files are available on the Open Science
Framework (DOI 10.17605/OSF.IO/AR8C3).
17Open asset ↗devoworm/Digital-Bacillariapdf-layout-page:17 lines:1-49Code · publicckground
color (select the background by color) to RGB value 0,0,0. To create a thick skeleton from a thin
skeleton, select the thin skeleton by color and then select the border function. The border width
should be set to 4, hard border, and filled with RGB value 0,217,0. The pseudo-code for GIMP
script-fu is located on Github (https://github.com/devoworm/Digital-Bacillaria/tree/master/Image-Skeletons).Image Tracking for Movement. We also employ image tracking for the primary microscopy
data. The tracking of a partial image (template) of a diatom can be used under certain conditions
to obtain its trajectory. In particular, a movement of the diatoms in a plane perpendicular to the
optical axiOpen asset ↗devoworm/Digital-Bacillariapdf-raw-page:10 lines:1-44Code · publicn-source software with a web interface called OpenDevoCell (based on DeepLearning 4J).
DeepLabv3 (Google, MountainView, California, USA) is a package for TensorFlow, and Deep
Learning 4J (Eclipse Foundation, Ottawa, Canada), a Java-based library that works with
TensorFlow. OpenDevoCell is open-source software located on Github
(https://github.com/devoworm/GSOC-2019/tree/master/OpenDevoCell) and as a web-based
application (https://open-devo-cell.herokuapp.com).11
.
CC-BY 4.0 International license
available under a
was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made
The copyright holder for this preprint (wOpen asset ↗devoworm/GSOC-2019pdf-raw-page:11 lines:1-35Code / dataset availability confirmedEurope PMC · checked 9 Sept 2026
Premise Physiological processes may vary within leaf laminae; however, the accompanying heterogeneity in leaf venation is rarely investigated because its quantification can be time consuming. Here we introduce accelerated protocols using existing software to increase sample throughput and ask whether laminae venation varies among three crop types and four subspecies of Brassica rapa . Methods FAA (formaldehyde, glacial acetic acid, and ethanol)-fixed samples were stored in ethanol. Without performing any additional clearing or staining, we tested two methods of image acquisition at three locations along the proximal-distal axis of the laminae and estimated the patterns of venation using the program phenoVein. We developed and made available an R script to handle the phenoVein output and then analyzed our data using linear mixed-effects models. Results Beyond fixation and storage, staining and clearing are not necessary to estimate leaf venation using phenoVein if the images are acquired using a stereomicroscope. All estimates of venation required some manual adjustment. We found a significant effect of location within the laminae for all aspects of venation. Discussion By removing the clearing and staining steps and utilizing the semi-automated program phenoVein, we quickly and cheaply acquired leaf venation data. Venation may be an important target for crop breeding efforts, particularly if intralaminar variation correlates with variation in physiological processes, which remains an open question.
Why it matches plant phenotyping methods葉脈という植物形態形質の画像取得・半自動定量法を開発・評価し、解析用Rスクリプトも提供しており、フェノタイピング手法が研究の中心である。
abstractHere we introduce accelerated protocols using existing software to increase sample throughput
Reproduction assets foundThe authors explicitly state that their custom R script for processing phenoVein output and all analyzed venation data are freely available on their public GitHub repository, making this a paper-specific, public, actionable asset.Code · publicWe developed an R script that (1) compiles data from multiple phenoVein .csv output files, (2) reformats the phenoVein output into a rectangular dataframe to facilitate the downstream data analysis, and (3) saves this new dataframe as a separate .csv file that can be easily imported into and analyzed using any number of statistical software environments (available at https://github.com/rlbaker5/AppsInPlantSci_phenoVein )Open asset ↗rlbaker5/AppsInPlantSci_phenoVeinlines:113-120Dataset · publicAll the analyzed data are available at https://github.com/rlbaker5/AppsInPlantSci_phenoVein .Open asset ↗rlbaker5/AppsInPlantSci_phenoVeinlines:113-120Code / dataset availability confirmedEurope PMC · checked 9 Sept 2026
Plasmodesmata are small channels that connect plant cells. While recent technological advances have facilitated the analysis of the ultrastructure of these channels, there are limitations to efficiently addressing their presence over an entire cellular interface. Here, we highlight the value of serial block electron microscopy for this purpose. We developed a computational pipeline to study plasmodesmata distributions and we detect presence/absence of plasmodesmata clusters, pit fields, at the phloem unloading interfaces of Arabidopsis thaliana roots. Pit fields can be visualised and quantified. As the wall environment of plasmodesmata is highly specialised we also designed a tool to extract the thickness of the extracellular matrix at and outside plasmodesmata positions. We show and quantify clear wall thinning around plasmodesmata with differences between genotypes, namely in the recently published plm-2 sphingolipid mutant. Our tools open new avenues for quantitative approaches in the analysis of symplastic trafficking. Sentence summary We developed computational tools for serial block electron microscopy datasets to extract information on the spatial distribution of plasmodesmata over an entire cellular interface and on the wall environment the plasmodesmata are in.
Why it matches plant phenotyping methods植物組織の電子顕微鏡画像から原形質連絡の分布や細胞壁厚を定量抽出する計算ツールとパイプラインが研究の中心であり、植物形態状態の測定法に該当する。
abstractWe developed a computational pipeline to study plasmodesmata distributions
Reproduction assets foundThe paper publicly releases its authors' MIB plugins for plasmodesmata distribution and wall-thickness analysis (GitHub), a guided R analysis tutorial/pipeline (GitHub Pages), and the Col-0 SB-EM datasets with segmented wall models and PD annotations (Google Drive), all with explicit availability statements and URLs.Code · publicA guided tutorial with all the necessary
code for this analysis is available at https://andreapaterlini.github.io/Plasmodesmata_dist_wall/Open asset ↗pdf-page:6 lines:1-49Dataset · publicThe Col-0 datasets used in this paper, with corresponding models and
annotation are available from https://drive.google.com/file/d/1g-Open asset ↗pdf-page:6 lines:1-49Code / dataset availability confirmedEurope PMC · checked 15 Sept 2026
The micro-morphology of leaf epidermises is valuable for the study of leaf development and function, as well as the classification of plant species. There have been few studies comparing different preparation and imaging methods for visualizing the leaf epidermis. Here, four specimen preparation methods were used to investigate the leaf epidermis morphology of Arabidopsis , radish, cucumber, wheat, rice, and maize, under an inverted basic light microscope (LM), a laser scanning confocal microscope (LSCM), or a scanning electron microscope (SEM). Optical microscope specimens were obtained using either the direct isolation method or the chloral hydrate-based clearing method. SEM images were obtained using a standard stage for conventional dehydrated samples or a Coolstage for fresh tissue. Different parts of epidermis peels were well focused under the LM. Investigation of samples cleared by chloral hydrate is convenient and autofluorescence of cell walls can be detected in rice. The resolution of images of conventional SEM leaf samples was generally higher than the Coolstage images at the same magnification, whereas local collapse and shrinkage were observed in leaves with high water content when using the conventional method. However, stomatal apparatuses of Arabidopsis , cucumber, radish, and maize deformed and showed poor appearance when using the Coolstage. Moreover, we usually used glutaraldehyde as an SEM fixative when using t-butanol for freeze-drying, though methanol is considered a better fixative in recent studies. In addition, fresh samples were not stable on the Coolstage. Thus, we compared four different t-butanol freeze-drying methods and two Coolstage methods. The dimension and morphology of tissues were compared using the six different methods. The results indicate that methanol fixative obviously reduced shrinkage of SEM samples compared with glutaraldehyde and formaldehyde alcohol acetic acid (FAA) fixatives. The use of methanol and a graded series of steps improved the preservation of samples. Preparing samples with optimal cutting temperature compound and observing at -30°C helped to increase the stability of Coolstage samples. In summary, our results provide an overview of the shortcomings and merits of four different methods, and might provide some information about choosing an optimal method for visualizing epidermal morphology.
Why it matches plant phenotyping methods葉表皮形態の可視化について、複数の試料調製法・顕微鏡法を比較し、組織形態の保存性や画像品質を評価しており、植物形質取得法が研究の中心である。
abstractThere have been few studies comparing different preparation and imaging methods for visualizing the leaf epidermis.
Reproduction assets foundThe paper reports LM/LSCM/SEM imaging of leaf epidermises and shrinkage/stability measurements. No author analysis code, trained models, or external repository deposit is mentioned. The only paper-specific public asset is the article's Supplementary Material, which the authors state contains all data generated or analySupplement · publicgy Project of Henan Province (182102110234).
Conflict of Interest
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Supplementary Material
The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fpls.2020.00133/full#supplementary-material
Click here for additional data file.
Abbreviations
SEM, scanning electron microscope; LSCM, laser scanning confocal microscope; LM, light microscope; DIC, differential interference contrast; CPD, critical point drying; OCT, optimum cutting temperature.
References
Bailes E. J. GlovOpen asset ↗lines:293-368Code / dataset availability confirmedCrossref · checked 15 Sept 2026
Miscanthus x giganteus (Mxg) is a promising second-generation biofuel crop with high production of energetic biomass. Our aim was to determine the level of plant stress of Mxg grown in poor quality soils using non-invasive physiological parameters and to test whether the stress could be reduced by application of plant growth regulators (PGRs). Plant fitness was quantified by measuring of leaf fluorescence using 24 indexes to select the most suitable fluorescence indicators for quantification of this type of abiotic stress. Simultaneously, visible stress signs were observed on stems and leaves and differences in variants were revealed also by microscopy of leaf sections. Leaf fluorescence analysis, visual observation and changes of leaf anatomy revealed significant stress in all studied subjects compared to those cultivated in good quality soil. Besides commonly used Fv/Fm (potential photosynthetic efficiency) and P.I. (performance index), which showed very low sensitivity, we suggest other fluorescence parameters (like dissipation, DIo/RC) for revealing finer differences. We can conclude that measurement of leaf fluorescence is a suitable method for revealing stress affecting Mxg in poor soils. However, none of investigated parameters proved significant positive effect of PGRs on stress reduction. Therefore, direct improvement of soil quality by fertilization should be considered for stress reduction and improving the biomass quality in this type of soils.
Why it matches plant phenotyping methods葉の蛍光指標を用いた非侵襲的ストレス定量と指標選定が研究目的の中心であり、植物の生理状態を測定するフェノタイピング手法の適用・検証に該当する。
abstractOur aim was to determine the level of plant stress of Mxg grown in poor quality soils using non-invasive physiological parameters
Reproduction assets foundThe paper's supplementary materials hosted on MDPI contain the paper-specific fluorescence index measurements (Table S1 means/SDs for all PGR concentrations, boxplots, experiment photos, climate data), which directly reproduce this study's plant-phenotyping measurements. No author analysis code or trained models are deSupplement · publics established that application of PGRs Stimpo and Regoplant did not reduce the stress level of Mxg, the direct improvement of soil shall be considered for stress reduction.
Acknowledgments
We would like to thank Agrobiotech for providing us with Stimpo and Regoplant.
Supplementary Materials
The following are available online at https://www.mdpi.com/2223-7747/9/2/194/s1 , Table S1: Means and standard deviations of fluorescence indexes for all PGRs concentrations in experiment; Figure S2: Boxplots of fluorescence indexes; Figure S3: Photograph of the experiment; Figure S4: Average month temperatures, precipitation and light period in Ústí nad Labem in 2017.
Click here for additional data file.Open asset ↗lines:96-146Code / dataset availability confirmedEurope PMC · Crossref · bioRxiv · checked 9 Sept 2026
ABSTRACT Quantitative analysis of plant and animal morphogenesis requires accurate segmentation of individual cells in volumetric images of growing organs. In the last years, deep learning has provided robust automated algorithms that approach human performance, with applications to bio-image analysis now starting to emerge. Here, we present PlantSeg, a pipeline for volumetric segmentation of plant tissues into cells. PlantSeg employs a convolutional neural network to predict cell boundaries and graph partitioning to segment cells based on the neural network predictions. PlantSeg was trained on fixed and live plant organs imaged with confocal and light sheet microscopes. PlantSeg delivers accurate results and generalizes well across different tissues, scales, and acquisition settings. We present results of PlantSeg applications in diverse developmental contexts. PlantSeg is free and open-source, with both a command line and a user-friendly graphical interface.
Why it matches plant phenotyping methods植物組織を細胞単位で3Dセグメンテーションする画像解析パイプラインを開発し、異なる組織・スケール・撮像条件で検証しているため、植物フェノタイピング手法が中心である。
abstractHere, we present PlantSeg, a pipeline for volumetric segmentation of plant tissues into cells.
Reproduction assets foundThe paper (PlantSeg) publicly releases its plant phenotyping inputs and analysis: raw confocal/light-sheet images with hand-curated groundtruth segmentations on OSF, and the open-source PlantSeg pipeline including pre-trained networks and evaluation scripts on GitHub.Dataset · publicPlantSeg is open-source and publicly available https://github.com/hci-unihd/plant-seg. The repository
includes a complete user guide, the evaluation scripts used for quantitative analysis, and the employed datasets.Open asset ↗github · hci-unihd/plant-segpdf-page:7 lines:1-45Code / dataset availability confirmedEurope PMC · Crossref · OpenAlex · checked 15 Sept 2026
Bulliform cells comprise specialized cell types that develop on the adaxial (upper) surface of grass leaves, and are patterned to form linear rows along the proximodistal axis of the adult leaf blade. Bulliform cell patterning affects leaf angle and is presumed to function during leaf rolling, thereby reducing water loss during temperature extremes and drought. In this study, epidermal leaf impressions were collected from a genetically and anatomically diverse population of maize inbred lines. Subsequently, convolutional neural networks were employed to measure microscopic, bulliform cell-patterning phenotypes in high-throughput. A genome-wide association study, combined with RNAseq analyses of the bulliform cell ontogenic zone, identified candidate regulatory genes affecting bulliform cell column number and cell width. This study is the first to combine machine learning approaches, transcriptomics, and genomics to study bulliform cell patterning, and the first to utilize natural variation to investigate the genetic architecture of this microscopic trait. In addition, this study provides insight toward the improvement of macroscopic traits such as drought resistance and plant architecture in an agronomically important crop plant.
Why it matches plant phenotyping methodsCNNを用いてトウモロコシ葉の微細なブルフォーム細胞形態を高スループット測定しており、表現型取得・抽出法が研究の中心です。
abstractSubsequently, convolutional neural networks were employed to measure microscopic, bulliform cell-patterning phenotypes in high-throughput.
Reproduction assets foundThe paper's Data Availability statement explicitly deposits the leaf epidermal glue-impression images (Cyverse zip), the authors' analysis scripts (LD calculation, image processing, U-net architecture, GWAS) on GitHub, trained U-net models (Cyverse File S1 zip), and supplemental material on figshare. All are paper-phenCode · publicwere deposited at NCBI SRT with SRA accession numbers PRJNA545465 and PRJNA400334. Leaf epidermal glue-impression images can be found at https://de.cyverse.org/dl/d/8CA8D72B-24AF-4887-8899-14460021887A/resized.zip. The scripts including LD calculation, image processing, U-net architecture, and running the GWAS are deposited in https://github.com/pengfei-qiao/Bulliform-cell-deep-learning.git. Trained U-net models are deposited as File S1 under https://de.cyverse.org/dl/d/B352A862-5B08-4373-87EB-9B48356028C6/FlieS1.zip. We request that this manuscript be cited when using these data. Supplemental material available at figshare: https://doi.org/10.25387/g3.9939623.Open asset ↗GitHub · pengfei-qiao/Bulliform-cell-deep-learninghtml-lines:236-236Code / dataset availability confirmedbioRxiv · checked 9 Sept 2026
The aerial epidermis of plants plays a major role in their environment interactions, and the development of its cellular components -trichomes, stomata and pavement cells- is still not fully understood. We have performed a detailed screen of the leaf epidermis of two generations of the well-established Solanum pennellii ac. LA716 x Solanum lycopersicum cv. M82 introgression line (IL) population using a combination of scanning electron microscopy techniques. Quantification of the trichome and stomatal densities in the ILs revealed 18 genomic regions with a low trichome density and 4 ILs with a high stomatal density. We also found ILs with abnormal proportions of different trichome types and aberrant trichome morphologies. This work has led to the identification of new, unexplored genomic regions with roles in trichome and stomatal formation and provides an important dataset for further studies on tomato epidermal development that is publically available to the research community.
Why it matches plant phenotyping methods走査電子顕微鏡を用いた葉表皮の画像取得と、毛状突起・気孔密度および形態の定量が研究の中心であり、再利用可能な表現型データセットも提供しているため。
abstractWe have performed a detailed screen of the leaf epidermis of two generations of the well-established Solanum pennellii ac. LA716 x Solanum lycopersicum cv. M82 introgression line (IL) population using a combination of scanning electron microscopy techniques.
Reproduction assets foundThe paper states that all SEM micrographs used for the trichome/stomatal phenotyping screen are publicly available in the BioStudies database under accession S-BSST262. This is a paper-specific public asset (the SEM images underlying the phenotyping measurements). No author analysis code was deposited.Dataset · public321 study are available in the BioStudies database (http://www.ebi.ac.uk/biostudies) (McEntyre et al.,Open asset ↗BioStudiespdf-page:10 lines:1-44Code / dataset availability confirmedOpenAlex · Europe PMC · checked 15 Sept 2026
Plant phenotyping to date typically comprises morphological and physiological profiling in a high-throughput manner. A powerful method that allows for subcellular characterization of organelle stoichiometric/functional characteristics is still missing. Organelle abundance and crosstalk in cell dynamics and signaling plays an important role for understanding crop growth and stress adaptations. However, microscopy can not be considered a high-throughput technology. The aim of the present study was to develop an approach that enables the estimation of organelle functional stoichiometry and to determine differential subcellular dynamics within and across cultivars in a high-throughput manner. A combination of subcellular non-aqueous fractionation and liquid chromatography mass spectrometry was applied to assign membrane-marker proteins to cell compartmental abundances and functions of Pisum sativum leaves. Based on specific subcellular affiliation, proteotypic marker peptides of the chloroplast, mitochondria and vacuole membranes were selected and synthesized as heavy isotope labelled standards. The rapid and unbiased Mass Western approach for accurate stoichiometry and targeted absolute protein quantification allowed for a proportional organelle abundances measure linked to their functional properties. A 3D Confocal Laser Scanning Microscopy approach was developed to evaluate the Mass Western. Two P. sativum cultivars of varying morphology and physiology were compared. The Mass Western assay enabled a cultivar specific discrimination of the chloroplast to mitochondria to vacuole relations.
Why it matches plant phenotyping methods植物の細胞内オルガネラ量と機能的特性を高スループットに推定するフェノタイピング手法を開発し、3D共焦点顕微鏡で評価・検証しているため、方法が中心的である。
abstractThe aim of the present study was to develop an approach that enables the estimation of organelle functional stoichiometry and to determine differential subcellular dynamics within and across cultivars in a high-throughput manner.
Reproduction assets foundThe paper's plant-phenotyping measurements and analysis outputs are available as public supplementary material hosted on the Frontiers article page: Tables S1–S4 (Mass Western target peptide lists, confocal organelle volume/area abundance values, NAF LFQ peak intensities, and proteotypic peptide subcellular localizatonSupplement · publicand Thomas Joch for plant cultivation at the department-associated greenhouse facility.
Footnotes
Funding. This study was funded by the Austrian Science Fund (FWF) [ P24870 -B22] and [W 1257-820], and supported by the COST action FA1306.
Supplementary Material
The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fpls.2019.00638/full#supplementary-material
Figure S1
Morphological phenotypes of the Pisum sativum cultivars Protecta (left) and Messire (right). Length of internodes and leaf weight n = 5 biol. replicates, error bars = standard error, p < 0.05 (Kruskal Wallis). ** p < 0.01, *** p < 0.005.
Click here for additional data filOpen asset ↗lines:102-128Code / dataset availability confirmedEurope PMC · checked 15 Sept 2026
Background Many methods have been developed to quantify cell shape in 2D in tissues. For instance, the analysis of epithelial cells in Drosophila embryogenesis or jigsaw puzzle-shaped pavement cells in plant epidermis has led to the development of numerous quantification methods that are applied to 2D images. However, proper extraction of 2D cell contours from 3D confocal stacks for such analysis can be problematic. Results We developed a macro in ImageJ, SurfCut, with the goal to provide a user-friendly pipeline specifically designed to extract epidermal cell contour signals, segment cells in 2D and analyze cell shape. As a reference point, we compared our output to that obtained with MorphoGraphX (MGX). While both methods differ in the approach used to extract the layer of signal, they output comparable results for tissues with shallow curvature, such as pavement cell shape in cotyledon epidermis (as quantified with PaCeQuant). SurfCut was however not appropriate for cell or tissue samples with high curvature, as evidenced by a significant bias in shape and area quantification. Conclusion We provide a new ImageJ pipeline, SurfCut, that allows the extraction of cell contours from 3D confocal stacks. SurfCut and MGX have complementary advantages: MGX is well suited for curvy samples and more complex analyses, up to computational cell-based modeling on real templates; SurfCut is well suited for rather flat samples, is simple to use, and has the advantage to be easily automated for batch analysis of images in ImageJ. The combination of these two methods thus provides an ideal suite of tools for cell contour extraction in most biological samples, whether 3D precision or high-throughput analysis is the main priority.
Why it matches plant phenotyping methods植物表皮細胞の輪郭・形状を3D画像から抽出・定量するImageJパイプラインの開発と比較検証が中心であり、植物形態フェノタイピング手法に該当する。
abstractWe developed a macro in ImageJ, SurfCut, with the goal to provide a user-friendly pipeline specifically designed to extract epidermal cell contour signals, segment cells in 2D and analyze cell shape.
Reproduction assets foundThe paper's authors publicly released both the SurfCut analysis macro (GitHub and Zenodo DOI 10.5281/zenodo.2635737) and the confocal microscopy dataset of plant samples used for the phenotyping measurements (Zenodo DOI 10.5281/zenodo.2577053).Code · publicDevo” and ERASMUS grant (20016-1-TR01-KA103-026029).
Availability of data and materials
The datasets generated and analyzed in this study are available in the Zenodo repository ( https://zenodo.org /), DOI:10.5281/zenodo.2577053 [ 34 ].
The script of the SurfCut macro and a more detailed step-by-step user guide are available at https://github.com/sverger/SurfCut [ 35 ], Zenodo DOI:10.5281/zenodo.2635737 [ 28 ].
Authors’ contributions
OE, ML, and SV performed the experiments. SV wrote the ImageJ script “SurfCut.” OE analyzed the results. OE, ML, OH, and SV wrote the article. OH secured funding for this project. All authors read and approved the final manuscript.
Ethics approval and consOpen asset ↗sverger/SurfCutlines:84-107Dataset · publicThe datasets generated and analyzed in this study are available in the Zenodo repository ( https://zenodo.org /), DOI:10.5281/zenodo.2577053 [ 34 ].Open asset ↗Zenodo · 10.5281/zenodo.2577053lines:84-107Code / dataset availability confirmedbioRxiv · Crossref · checked 15 Sept 2026
BackgroundOne of the main features of plant cells is their strong plasticity, and their propensity to regenerate an organism from a single cell. Plant protoplasts are basic plant cells units in which the pecto-cellulosic cell wall has been removed, but the plasma membrane is intact. One of the main features of plant cells is their strong plasticity, which in some species, can be very close from what is defined as cell totipotency. Methods and differentiation protocols used in plant physiology and plant biology usually involve macroscopic vessels and containers that make difficult, for example, to follow the fate of the same protoplast all along its full development cycle, but also to perform continuous studies of the influence of various gradients in this context. These limits have hampered the precise study of regeneration processes. ResultsHerein, we present the design of a comprehensive, physiologically relevant, easy-to-use and low-cost microfluidic and microscopic setup for the monitoring of Physcomitrella patens (P. patens) growth and development on a long-term basis. The experimental solution we developed is made of two parts (i) a microfluidic chip composed of a single layer of about a hundred flow-through microfluidic traps for the immobilization of protoplasts, and (ii) a low-cost, light-controlled, custom-made microscope allowing the continuous recording of the moss development in physiological conditions. We validated the experimental setup with three proofs of concepts: (i) the kinetic monitoring of first division steps and cell wall regeneration, (ii) the influence of the photoperiod on growth of the protonemata, and (iii) finally the induction of leafy buds using a phytohormone, cytokinin. ConclusionsWe developed the design of a comprehensive, physiologically relevant, easy-to-use and low-cost experimental setup for the study of P. patens development in a microfluidic environment. This setup allows imaging of P. patens development at high resolution and over long time periods.
Why it matches plant phenotyping methods植物の発生・成長を長期間画像モニタリングするマイクロ流体チップとカスタム顕微鏡を開発しており、表現型取得系が研究の中心である。
abstractwe present the design of a comprehensive, physiologically relevant, easy-to-use and low-cost microfluidic and microscopic setup for the monitoring of Physcomitrella patens (P. patens) growth and development on a long-term basis.
Reproduction assets foundThe paper's availability statement points to a public GitHub repository (FattaccioliLab/PlantsOnChip) containing the authors' microfluidic chip design files, microscope control Matlab scripts, Micromanager configuration, Arduino connection map, and bill of materials used for the plant protoplast phenotyping/imaging. NoCode · publicfile of the 35 mm Petri dish adapter to the SM1 threading of the xy manual stage
• Matlab programming script of the microscope and Micromanager configuration file
• Connection map of the Arduino Due board
• Bill of materials of the custom-made microscope (references, manufacturers, suppliers, prices)
Documents are available on https://github.com/FattaccioliLab/PlantsOnChip
Supplementary movies.
• Division of a protoplast and cell wall regeneration kinetics
• Chloronemata growth under continuous illumination
Competing interests. No financial competing interests are to be declared.
Funding. This work has received support of “Institut Pierre-Gilles de Gennes” (Laboratoire
d’excellence : ANR-10-Open asset ↗FattaccioliLab/PlantsOnChippdf-raw-page:11 lines:1-29Code / dataset availability confirmedEurope PMC · checked 10 Sept 2026
Stomata control gas exchanges between the plant and the atmosphere. How natural variation in stomata size and density contributes to resolve trade-offs between carbon uptake and water loss in response to local climatic variation is not yet understood. We developed an automated confocal microscopy approach to characterize natural genetic variation in stomatal patterning in 330 fully sequenced Arabidopsis thaliana accessions collected throughout the European range of the species. We compared this to variation in water-use efficiency, measured as carbon isotope discrimination (δ 13 C). We detect substantial genetic variation for stomata size and density segregating within Arabidopsis thaliana. A positive correlation between stomata size and δ 13 C further suggests that this variation has consequences on water-use efficiency. Genome wide association analyses indicate a complex genetic architecture underlying not only variation in stomatal patterning but also to its covariation with carbon uptake parameters. Yet, we report two novel QTL affecting δ 13 C independently of stomatal patterning. This suggests that, in A. thaliana, both morphological and physiological variants contribute to genetic variance in water-use efficiency. Patterns of regional differentiation and covariation with climatic parameters indicate that natural selection has contributed to shape some of this variation, especially in Southern Sweden, where water availability is more limited in spring relative to summer. These conditions are expected to favour the evolution of drought avoidance mechanisms over drought escape strategies.
Why it matches plant phenotyping methods自動化共焦点顕微鏡による気孔サイズ・密度の表現型取得法を開発し、330系統へ大規模適用しているため、植物表現型計測が中心である。
abstractWe developed an automated confocal microscopy approach to characterize natural genetic variation in stomatal patterning in 330 fully sequenced Arabidopsis thaliana accessions collected throughout the European range of the species.
Reproduction assets foundThe paper's Data Accessibility statement explicitly deposits raw confocal image data, image analysis scripts, and phenotypic data in a Dryad repository, uploads genotypic phenotype means to AraPheno, and provides authors' GWAS and MTMM analysis scripts on GitHub. All are paper-specific, public, and actionable.Dataset · publicRaw image data and image analysis scripts are stored in a Dryad repository ( https://doi.org/10.5061/dryad.n068q74 ). Phenotypic data are provided as supplemental material and included in the Dryad repository.Open asset ↗Dryad · 10.5061/dryad.n068q74lines:153-213Code · publicGWAS scripts are available at https://github.com/arthurkorte/GWAS .Open asset ↗GitHub · arthurkorte/GWASlines:153-213Code · publicMTMM scripts are available at https://github.com/Gregor-Mendel-Institute/mtmm .Open asset ↗GitHub · Gregor-Mendel-Institute/mtmmlines:153-213Code / dataset availability confirmedbioRxiv · Europe PMC · checked 15 Sept 2026
Stomata control gas exchanges between the plant and the atmosphere. How natural variation in stomata size and density contributes to resolve trade-offs between carbon uptake and water-loss in response to local climatic variation is not yet understood. We developed an automated confocal microscopy approach to characterize natural genetic variation in stomatal patterning in 330 fully-sequenced Arabidopsis thaliana accessions collected throughout the European range of the species. We compared this to variation in water-use efficiency, measured as carbon isotope discrimination ({delta}13C). We detect substantial genetic variation for stomata size and density segregating within Arabidopsis thaliana. A positive correlation between stomata size and {delta}13C further suggests that this variation has consequences on water-use efficiency. Genome-wide association analyses indicate a complex genetic architecture underlying not only variation in stomata patterning but also to its co-variation with carbon uptake parameters. Yet, we report two novel QTL affecting {delta}13C independently of stomata patterning. This suggests that, in A. thaliana, both morphological and physiological variants contribute to genetic variance in water-use efficiency. Patterns of regional differentiation and co-variation with climatic parameters indicate that natural selection has contributed to shape some of this variation, especially in Southern Sweden, where water availability is more limited in spring relative to summer. These conditions are expected to favor the evolution of drought avoidance mechanisms over drought escape strategies.
Why it matches plant phenotyping methods自動共焦点顕微鏡法を開発し、330系統で気孔サイズ・密度という植物形質を大規模に測定しており、表現型取得法が研究の中心です。
abstractWe developed an automated confocal microscopy approach to characterize natural genetic variation in stomatal patterning in 330 fully-sequenced Arabidopsis thaliana accessions collected throughout the European range of the species.
Reproduction assets foundThe data accessibility statement lists public, paper-specific assets: phenotypic (stomata/δ13C) data to be deposited in AraPheno with a public URL, and the authors' GWAS and MTMM analysis scripts on GitHub. Raw images and image-analysis scripts are only available upon request (Dryad deposit pending acceptance), so theyCode · public1001genomes.org, (Seren et
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al., 2017) and stored in a Dryad repository upon acceptance. Additionally, we provide an R
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Markdown file, which contains all figures (except GWAS and MTMM) and the
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corresponding R code used to create the figures and statistics in the supplemental material.
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GWAS scripts are available at https://github.com/arthurkorte/GWAS. MTMM scripts are
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available at https://github.com/Gregor-Mendel-Institute/mtmm.925
Genomic data used is publicly available in the 1001 genomes database (Alonso-Blanco et al.,
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2016)
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JdM, AK, and HD conceived the study. HD conducted the experiment and produced
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phenotypic data for stomaOpen asset ↗arthurkorte/GWASpdf-raw-page:35 lines:1-46Code · publicory upon acceptance. Additionally, we provide an R
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Markdown file, which contains all figures (except GWAS and MTMM) and the
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corresponding R code used to create the figures and statistics in the supplemental material.
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GWAS scripts are available at https://github.com/arthurkorte/GWAS. MTMM scripts are
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available at https://github.com/Gregor-Mendel-Institute/mtmm.925
Genomic data used is publicly available in the 1001 genomes database (Alonso-Blanco et al.,
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2016)
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JdM, AK, and HD conceived the study. HD conducted the experiment and produced
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phenotypic data for stomata traits. TM and AW were responsible for 13
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provideOpen asset ↗Gregor-Mendel-Institute/mtmm.925pdf-raw-page:35 lines:1-46Code / dataset availability confirmedbioRxiv · Europe PMC · checked 14 Sept 2026
O_LIStomata fulfill an important physiological role and are often phenotyped by researchers in many fields. Currently, no fully automated method exists to perform this task. Researchers typically rely on manual counts of stomata, which is an error-prone method and difficult to reproduce.\nC_LIO_LIWe introduce StomataCounter, an automated stomata counting system using a deep convolutional neural network to identify pores in a variety of different microscopic images. We used a human-in-the-loop approach to train and refine a neural network on a large variety of microscopic images, which helps us achieve robust detection among a number of datasets.\nC_LIO_LIOur network achieves 98.1% identification accuracy on Ginkgo SEM micrographs, and 94.2% transfer accuracy when tested on untrained species.\nC_LIO_LITo facilitate adoption of the method, we make a web tool available under http://www.stomata.science/\nC_LI
Why it matches plant phenotyping methods気孔という植物形質の画像ベース自動同定・計数法を開発し、異なる画像・種で精度検証した研究であり、方法自体が中心です。
abstractWe introduce StomataCounter, an automated stomata counting system using a deep convolutional neural network to identify pores in a variety of different microscopic images.
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Code · public125 codes for network training, as well as the webserver are available at http://stomata.science/source. To useOpen asset ↗pdf-page:5 lines:1-57Code / dataset availability confirmedCrossref · checked 15 Sept 2026
Most effective nematicides for the control of root-knot nematodes are banned, which demands a better understanding of the plant-nematode interaction. Understanding how gene expression in the nematode-feeding sites relates to morphological features may assist a better characterization of the interaction. However, nematode-induced galls resulting from cell-proliferation and hypertrophy hinders such observation, which would require tissue sectioning or clearing. We demonstrate that a method based on the green auto-fluorescence produced by glutaraldehyde and the tissue-clearing properties of benzyl-alcohol/benzyl-benzoate preserves the structure of the nematode-feeding sites and the plant-nematode interface with unprecedented resolution quality. This allowed us to obtain detailed measurements of the giant cells’ area in an Arabidopsis line overexpressing CHITINASE-LIKE-1 (CTL1) from optical sections by confocal microscopy, assigning a role for CTL1 and adding essential data to the scarce information of the role of gene repression in giant cells. Furthermore, subcellular structures and features of the nematodes body and tissues from thick organs formed after different biotic interactions, i.e., galls, syncytia, and nodules, were clearly distinguished without embedding or sectioning in different plant species (Arabidopsis, cucumber or Medicago). The combination of this method with molecular studies will be valuable for a better understanding of the plant-biotic interactions.
Why it matches plant phenotyping methods根こぶ線虫摂食部位の構造を共焦点画像から高解像度に取得し、巨大細胞面積を測定する植物フェノタイピング法の開発が中心である。
titleA Phenotyping Method of Giant Cells from Root-Knot Nematode Feeding Sites by Confocal Microscopy Highlights a Role for CHITINASE-LIKE 1 in Arabidopsis
Reproduction assets foundThe paper describes a confocal-microscopy phenotyping method for nematode-induced giant cells. The only paper-specific public asset referenced is the authors' supplementary material (hosted at MDPI), which per the text contains Table S1 (gene filtering results) and Videos S6–S9 of the confocal optical sections used forSupplement · public(PEII-2014-020-P to Carmen Fenoll). Javier Cabrera is supported by a Cytema-Santander contract from Universidad de Castilla-La Mancha. Christian Hermans is a research associate from Fonds de la Recherche Scientifique—National Fund for Scientific Research (Belgium).
Supplementary Materials
Supplementary materials can be found at http://www.mdpi.com/1422-0067/19/2/429/s1 and www.mdpi.com/1422-0067/19/2/429/s2 .
Click here for additional data file.
Click here for additional data file.
Author Contributions
Javier Cabrera, Rocio Olmo, Virginia Ruiz-Ferrer, and Christian Hermans conceived and designed the experiments; Javier Cabrera, Rocio Olmo, Virginia Ruiz-Ferrer, Christian Hermans, and Isabel Open asset ↗lines:51-66Code / dataset availability confirmedCrossref · checked 14 Sept 2026
Wheat (Triticum aestivum L.) is an important food crop, and biotic and abiotic stresses significantly impact grain yield. Wheat leaf and stem surface waxes are associated with traits of biological importance, including stress resistance. Past studies have characterized the composition of wheat cuticular waxes, however protocols can be relatively low-throughput and narrow in the range of metabolites detected. Here, gas chromatography-mass spectrometry (GC-MS) metabolomics methods were utilized to provide a comprehensive characterization of the chemical composition of cuticular waxes in wheat leaves and stems. Further, waxes from four wheat cultivars were assayed to evaluate the potential for GC-MS metabolomics to describe wax composition attributed to differences in wheat genotype. A total of 263 putative compounds were detected and included 58 wax compounds that can be classified (e.g., alkanes and fatty acids). Many of the detected wax metabolites have known associations to important biological functions. Principal component analysis and ANOVA were used to evaluate metabolite distribution, which was attributed to both tissue type (leaf, stem) and cultivar differences. Leaves contained more primary alcohols than stems such as 6-methylheptacosan-1-ol and octacosan-1-ol. The metabolite data were validated using scanning electron microscopy of epicuticular wax crystals which detected wax tubules and platelets. Conan was the only cultivar to display alcohol-associated platelet-shaped crystals on its abaxial leaf surface. Taken together, application of GC-MS metabolomics enabled the characterization of cuticular wax content in wheat tissues and provided relative quantitative comparisons among sample types, thus contributing to the understanding of wax composition associated with important phenotypic traits in a major crop.
Why it matches plant phenotyping methodsGC-MSメタボロミクスを用いた植物表面ワックス組成の包括的な取得・比較を主題とし、SEMによる検証も行っているため、化学的な植物形質の測定法として中心的です。
abstractHere, gas chromatography-mass spectrometry (GC-MS) metabolomics methods were utilized to provide a comprehensive characterization of the chemical composition of cuticular waxes in wheat leaves and stems.
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Supplement · publicThe following are available online at http://www.mdpi.com/1422-0067/19/2/249/s1 . Figure S1. Wax density.docx provides a semi-quantitative analysis of wheat epicuticular wax density using image processing tools on SEM micrographs; Table S1. Wax metabolite annotations.txt provides detailed information on detected metabolites.Open asset ↗lines:522-564Code / dataset availability confirmedEurope PMC · OpenAlex · checked 10 Sept 2026
Background Crop species are of increasing interest both for cattle feeding and for bioethanol production. The degradability of the plant material largely depends on the lignification of the tissues, but it also depends on histological features such as the cellular morphology or the relative amount of each tissue fraction. There is therefore a need for high-throughput phenotyping systems that quantify the histology of plant sections. Results We developed custom image processing and an analysis procedure for quantifying the histology of maize stem sections coloured with FASGA staining and digitalised with whole microscopy slide scanners. The procedure results in an automated segmentation of the input images into distinct tissue regions. The size and the fraction area of each tissue region can be quantified, as well as the average coloration within each region. The measured features can discriminate contrasted genotypes and identify changes in histology induced by environmental factors such as water deficit. Conclusions The simplicity and the availability of the software will facilitate the elucidation of the relationships between the chemical composition of the tissues and changes in plant histology. The tool is expected to be useful for the study of large genetic populations, and to better understand the impact of environmental factors on plant histology.
Why it matches plant phenotyping methodsトウモロコシ茎切片の組織形態を画像処理で自動分割・定量する手法を開発しており、植物表現型の取得・抽出が研究の中心である。
abstractWe developed custom image processing and an analysis procedure for quantifying the histology of maize stem sections coloured with FASGA staining and digitalised with whole microscopy slide scanners.
Reproduction assets foundThe paper's image segmentation/quantification workflow is publicly released as an ImageJ/Fiji plugin (QuantifFasga) on GitHub, and the authors' in-house Matlab statistical analysis library (MatStats) is also publicly available on GitHub. The phenotype measurement data themselves are only available upon request.Code · publicThe code for the segmentation of tissue regions and the quantification of histology is freely available on the Internet through the GitHub platform at http://github.com/ijpb/fasga-quantif/releases (last accessed: August 8, 2017).Open asset ↗ijpb/fasga-quantiflines:651-651Code / dataset availability confirmedEurope PMC · checked 10 Sept 2026
Vibrational spectroscopy provides non-destructively the molecular fingerprint of plant cells in the native state. In combination with microscopy, the chemical composition can be followed in context with the microstructure, and due to the non-destructive application, in-situ studies of changes during, e.g., degradation or mechanical load are possible. The two complementary vibrational microspectroscopic approaches, Fourier-Transform Infrared (FT-IR) Microspectroscopy and Confocal Raman spectroscopy, are based on different physical principles and the resulting different drawbacks and advantages in plant applications are reviewed. Examples for FT-IR and Raman microscopy applications on plant cell walls, including imaging as well as in-situ studies, are shown to have high potential to get a deeper understanding of structure-function relationships as well as biological processes and technical treatments. Both probe numerous different molecular vibrations of all components at once and thus result in spectra with many overlapping bands, a challenge for assignment and interpretation. With the help of multivariate unmixing methods (e.g., vertex components analysis), the most pure components can be revealed and their distribution mapped, even tiny layers and structures (250 nm). Instrumental as well as data analysis progresses make both microspectroscopic methods more and more promising tools in plant cell wall research.
Why it matches plant phenotyping methods植物細胞壁の構造・化学組成を対象とする振動顕微分光法（FT-IRおよび共焦点ラマン）の植物への応用、画像化、データ解析を方法論としてレビューしており、植物状態の取得・抽出法が中心である。
abstractThe two complementary vibrational microspectroscopic approaches, Fourier-Transform Infrared (FT-IR) Microspectroscopy and Confocal Raman spectroscopy, are based on different physical principles and the resulting different drawbacks and advantages in plant applications are reviewed.
Reproduction assets foundThe review mentions an author-established public spectral database of plant cell wall reference components and spectra, hosted at bionami.at/spectra.html, which directly supports the paper's vibrational microspectroscopy measurements and band-assignment analysis. No code, models, or image datasets with explicit public-Dataset · publica spectral database, including reference components as well as different plant cell walls is currently established and made available to the scientific community ( http://bionami.at/spectra.html ).Open asset ↗bionami.atlines:98-107Code / dataset availability confirmedEurope PMC · checked 10 Sept 2026
Pavement cells (PCs) are the most frequently occurring cell type in the leaf epidermis and play important roles in leaf growth and function. In many plant species, PCs form highly complex jigsaw-puzzle-shaped cells with interlocking lobes. Understanding of their development is of high interest for plant science research because of their importance for leaf growth and hence for plant fitness and crop yield. Studies of PC development, however, are limited, because robust methods are lacking that enable automatic segmentation and quantification of PC shape parameters suitable to reflect their cellular complexity. Here, we present our new ImageJ-based tool, PaCeQuant, which provides a fully automatic image analysis workflow for PC shape quantification. PaCeQuant automatically detects cell boundaries of PCs from confocal input images and enables manual correction of automatic segmentation results or direct import of manually segmented cells. PaCeQuant simultaneously extracts 27 shape features that include global, contour-based, skeleton-based, and PC-specific object descriptors. In addition, we included a method for classification and analysis of lobes at two-cell junctions and three-cell junctions, respectively. We provide an R script for graphical visualization and statistical analysis. We validated PaCeQuant by extensive comparative analysis to manual segmentation and existing quantification tools and demonstrated its usability to analyze PC shape characteristics during development and between different genotypes. PaCeQuant thus provides a platform for robust, efficient, and reproducible quantitative analysis of PC shape characteristics that can easily be applied to study PC development in large data sets.
Why it matches plant phenotyping methods葉表皮の舗装細胞形状を画像から自動抽出・定量するImageJツールを開発し、手動セグメンテーション等との比較検証も行っており、植物フェノタイピング手法が中心である。
abstractHere, we present our new ImageJ-based tool, PaCeQuant, which provides a fully automatic image analysis workflow for PC shape quantification.
Reproduction assets foundThe paper's PaCeQuant image analysis tool (ImageJ/MiToBo plugin) and its source code are publicly available under GPL v3.0 via the MiToBo website and GitHub, directly implementing the paper's pavement cell segmentation and 27-feature quantification workflow. The R analysis script is only described as supplemental, and Code · publicThe source code of MiToBo and the PaCeQuant plugin is available from the MiToBo website or on Github ( https://github.com/mitobo-hub/mitobo )Open asset ↗mitobo-hub/mitobolines:95-99Code / dataset availability confirmedbioRxiv · Europe PMC · checked 14 Sept 2026
How complex developmental-genetic networks are translated into organs with specific 3D shapes remains an open question. This question is particularly challenging because the elaboration of specific shapes is in essence a question of mechanics. In plants, this means how the genetic circuitry affects the cell wall. The mechanical properties of the wall and their spatial variation are the key factors controlling morphogenesis in plants. However, these properties are difficult to measure and investigating their relation to genetic regulation is particularly challenging. To measure spatial variation of mechanical properties, one must determine the deformation of a tissue in response to a known force with cellular resolution. Here we present an automated confocal micro-extensometer (ACME), which greatly expands the scope of existing methods for measuring mechanical properties. Unlike classical extensometers, ACME is mounted on a confocal microscope and utilizes confocal images to compute the deformation of the tissue directly from biological markers, thus providing cellular scale information and improved accuracy. ACME is suitable for measuring the mechanical responses in live tissue. As a proof of concept we demonstrate that the plant hormone gibberellic acid induces a spatial gradient in mechanical properties along the length of the Arabidopsis hypocotyl.\n\nTerms
Why it matches plant phenotyping methods植物組織の力学的性質を細胞解像度で定量する自動共焦点マイクロ伸展計を開発し、画像から変形を抽出する方法を中心に実証しているため。
abstractHere we present an automated confocal micro-extensometer (ACME), which greatly expands the scope of existing methods for measuring mechanical properties.
Reproduction assets found保存済みの本文根拠を更新済みルールで再検証し、公開資産1件を確認しました。Code · publicThe positioners are controlled by a SmarAct MCS3D (SmarAct GmbH) controller (Figure 1C,
label 15) accompanied by its software library, which in turn is controlled by custom-made software
(available here: https://github.com/ACME-Robinson/InstallPackage)Open asset ↗ACME-Robinson/InstallPackagepdf-page:14 lines:1-54Code / dataset availability confirmedEurope PMC · checked 15 Sept 2026
Root-knot nematodes induce galls that contain giant-feeding cells harboring multiple enlarged nuclei within the roots of host plants. It is recognized that the cell cycle plays an essential role in the set-up of a peculiar nuclear organization that seemingly steers nematode feeding site induction and development. Functional studies of a large set of cell cycle genes in transgenic lines of the model host Arabidopsis thaliana have contributed to better understand the role of the cell cycle components and their implication in the establishment of functional galls. Mitotic activity mainly occurs during the initial stages of gall development and is followed by an intense endoreduplication phase imperative to produce giant-feeding cells, essential to form vigorous galls. Transgenic lines overexpressing particular cell cycle genes can provoke severe nuclei phenotype changes mainly at later stages of feeding site development. This can result in chaotic nuclear phenotypes affecting their volume. These aberrant nuclear organizations are hampering gall development and nematode maturation. Herein we report on two nuclear volume assessment methods which provide information on the complex changes occurring in nuclei during giant cell development. Although we observed that the data obtained with AMIRA tend to be more detailed than Volumest (Image J), both approaches proved to be highly versatile, allowing to access 3D morphological changes in nuclei of complex tissues and organs. The protocol presented here is based on standard confocal optical sectioning and 3-D image analysis and can be applied to study any volume and shape of cellular organelles in various complex biological specimens. Our results suggest that an increase in giant cell nuclear volume is not solely linked to increasing ploidy levels, but might result from the accumulation of mitotic defects.
Why it matches plant phenotyping methods根こぶ線虫誘導巨大細胞の核体積・3D形態を取得する画像解析手法を提示し、AMIRAとVolumestを比較検証しているため、植物表現型計測法が中心です。
abstractHerein we report on two nuclear volume assessment methods which provide information on the complex changes occurring in nuclei during giant cell development.
Reproduction assets foundThe paper's nuclear volume measurements (individual GC and NGC nuclear volumes for Col-0, KRP3 OE, and KRP5 OE lines) are deposited in the article's public Supplementary File 1, available via the Frontiers supplementary-material URL. The Volumest plugin URL is a generic third-party tool, not a paper-specific asset, andSupplement · publicS-COFECUB (n°. sv 683/10 2011) project. RC has been supported by a doctoral scholarship in Brazil from CNPq (process number: 143030/2009-4) and in France from CAPES (process number: 6585-11-6).
1
http://lepo.it.da.ut.ee/~markkom/volumest/
Supplementary Material
The Supplementary Material for this article can be found online at: http://journal.frontiersin.org/article/10.3389/fpls.2017.00961/full#supplementary-material
Click here for additional data file.
References
Banora M. Y. Rodiuc N. Baldacci-Cresp F. Smertenko A. Bleve-Zacheo T. Mellilo M. T.
( 2011 ).
Feeding cells induced by phytoparasitic nematodes require gamma-tubulin ring complex for microtubule reorganization.
PLoS Pathog.
7 : e10Open asset ↗lines:88-182