Impedance flow cytometry for rapid quality assessment of protoplast cultures.

Abstract

Background Protoplasts, which are plant cells devoid of cell walls, are valuable tools in plant biotechnology. However, they are highly sensitive to mechanical and osmotic stress during isolation and early culture, often leading to significant loss of viability. Reliable and efficient methods for monitoring protoplast quality are essential for downstream applications. Results We applied impedance flow cytometry to assess the viability, cell size, and early division of freshly isolated protoplasts from Arabidopsis thaliana, Brassica napus, and Beta vulgaris. This label-free technique enables fast, objective, and high-throughput assessment of individual protoplasts, allowing reliable monitoring of viability and early division in large populations. Importantly, IFC-derived viability metrics strongly correlated with microcallus formation, demonstrating their predictive value for culture competence. Conclusions Impedance flow cytometry provides a robust, efficient and reproducible method for characterizing protoplast cultures. It enables rapid assessment of viability and growth potential, supporting quality control and optimization in plant cell culture workflows.

Code and data availability

The supplied blocks describe IFC measurements, microscopy images, and R-based analysis, but contain no data availability statement, no public repository deposit, and no author code URL. The only supplementary item is a DOCX with no stated contents matching qualifying assets.

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