Achieving Micrometer-Scale 4D X-ray tomography of Living Leaf Tissue in the Laboratory

Abstract

A methodology for achieving micrometer-scale 4D X-ray lab microscopy of living leaf tissue was developed to overcome challenges associated with delicate tissues, radiation damage, and motion artifacts during in vivo imaging. The study focused on optimizing laboratory based X-ray micro-computed tomography (microCT) parameters to balance high-resolution imaging with minimized physiological stress and radiation dose quantification. Assessing the dose-safe imaging window required comparing vertical and horizontal leaf mounting setups. Results demonstrated that the horizontal setup provided greater stability, preventing tissue degradation and maintaining sample viability during continuous acquisitions lasting up to 22 hours ([~]15600 Gy). MicroCT capacities were clearly able to resolve microstructures at the cellular level, achieving a pixel size down to 1 {micro}m. Furthermore, this optimized methodology confirmed the ability to track the spatiotemporal dynamics of applied compounds such as iohexol and aggregated nanoparticles within the leaf tissue. This work establishes that accessible laboratory based microCT enables the in vivo 4D monitoring of anatomical and physiological changes in living plants.

Plant phenotyping relevance

生葉を対象とした高解像度4D X線マイクロCTの撮像条件・線量・試料配置を開発し、生体内の解剖学的・生理学的変化を追跡する方法が研究の中心である。

Code and data availability

The supplied blocks describe lab-based microCT phenotyping of living barley/grass leaves (dose quantification, stomatal imaging, NP/contrast-agent tracking) but contain no public phenotype dataset, image deposit, author code, or trained model. The only URLs are citations, commercial software (Dragonfly, Excillum), and

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