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Background <p>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.</p> Results <p>We applied impedance flow cytometry to assess the viability, cell size, and early division of freshly isolated protoplasts from <i>Arabidopsis thaliana</i>,<i> Brassica napus</i>, and <i>Beta vulgaris</i>. 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.</p> Conclusions <p>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.</p>

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Impedance flow cytometry for rapid quality assessment of protoplast cultures

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年12月9日

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  • Tiago Rodrigues,
  • Robin Lardon,
  • Mária Šimášková,
  • Hilde Van Houtte,
  • Shivegowda Thammannagowda,
  • Grit Schade,
  • Steffen Vanneste,
  • Danny Geelen

摘要

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.