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High-throughput quantitative analysis of mitotic defects in fission yeast using Imaging Flow Cytometry
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  • Martin Prevorovsky,
  • Viacheslav Zemlianski,
  • Jarmila Princová,
  • Martin Báječný
Martin Prevorovsky
Univerzita Karlova Prirodovedecka fakulta
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Viacheslav Zemlianski
Univerzita Karlova Prirodovedecka fakulta

Corresponding Author:[email protected]

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Jarmila Princová
Univerzita Karlova Prirodovedecka fakulta
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Martin Báječný
Univerzita Karlova 1 lekarska fakulta
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Abstract

In this study, we introduce a novel approach for the high-throughput quantitative analysis of mitotic defects in the fission yeast Schizosaccharomyces pombe using Imaging Flow Cytometry (IFC). Fission yeast is a valuable model organism for cell cycle research, and the ‘cut’ phenotype, indicative of mitotic catastrophe, has been instrumental in discovering cell cycle regulators. Traditional fluorescence microscopy methods for quantifying ‘cut’ events suffer from subjectivity and limited throughput. Our IFC pipeline overcomes these limitations by automating the detection of ‘cut’ cells based on the unique characteristic of daughter cell nuclei becoming trapped in the cell wall during aberrant mitosis. We demonstrate the pipeline’s effectiveness using wild-type and mutant strains, with results validated against manual scoring. Our study establishes IFC as a powerful tool for investigating mitotic fidelity in fission yeast, with implications for advancing cell biology research.
16 Oct 2023Submitted to Yeast
16 Oct 2023Submission Checks Completed
16 Oct 2023Assigned to Editor
16 Oct 2023Review(s) Completed, Editorial Evaluation Pending
26 Oct 2023Reviewer(s) Assigned