Spectral algal fingerprinting and long sequencing in synthetic algal-microbial communities

Abstract

O_LISynthetic biology has made progress in creating artificial microbial and algal communities, but technical and evolutionary complexities still pose significant challenges. C_LIO_LITraditional methods for studying microbial and algal communities, such as microscopy and pigment analysis, are limited in throughput and resolution. In contrast, advancements in full-spectrum cytometry enabled high-throughput, multidimensional analysis of single cells based on their size, complexity, and spectral fingerprints, offering more precise and comprehensive analysis than conventional flow cytometry. C_LIO_LIThis study demonstrates the use of full-spectrum cytometry for analyzing synthetic algal-microbial communities, facilitating rapid species identification and enumeration. The workflow involves recording individual spectral signatures from monocultures, utilizing autofluorescence to distinguish them from noise, and subsequent creation of a spectral library for further analysis. The obtained library is used then to analyze mixtures of unicellular cyanobacteria and synthetic phytoplankton communities, revealing differences in spectral signatures. The synthetic consortium experiment monitored algal growth, comparing results from different instruments and highlighting the advantages of the spectral virtual filter system for precise population separation and abundance tracking. This approach demonstrated higher flexibility and accuracy in analyzing multi-component algal-microbial assemblages and tracking temporal changes in community composition. C_LIO_LIBy capturing the complete emission spectrum of each cell, this method enhances the understanding of algal-microbial community dynamics and responses to environmental stressors. With development of standardized spectral libraries, our work demonstrates an improved characterization of algal communities, advancing research in synthetic biology and phytoplankton ecology. C_LI

Plant phenotyping relevance

藻類の個体スペクトル計測とスペクトルライブラリを用いて、群集の構成・個体数・増殖を高スループットに測定する技術が研究の中心であり、植物状態の取得法として実質的です。

Code and data availability

The paper's spectral cytometry data (algal spectral signatures, gating/VF analyses) are paper-specific but not publicly deposited; they are available only from the corresponding author on request. The only public deposit is the 16S rRNA sequencing data (NCBI SRA PRJNA1131605), which is a molecular omics deposit and not

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