Fluorescence exclusion - a rapid, accurate and powerful method for measuring yeast cell volume

Area of Science:

• Cell Biology
• Microbiology
• Biophysics

Background:

• Cell volume varies significantly across species and within species.
• Accurate cell volume determination is challenging, limiting understanding of cell size control.
• Current methods often rely on indirect measurements, especially in yeast models.

Purpose of the Study:

• To assess the applicability of the fluorescence exclusion method (FXm) for measuring the volume of smaller cells, specifically fission yeast.
• To evaluate FXm's ability to detect subtle volume differences and dynamic changes in yeast cells.
• To enable new insights into cell growth biology through the combination of FXm and yeast genetics.

Main Methods:

• Development of specialized microfluidic chips for yeast cell analysis.
• Implementation of an improved data analysis pipeline for FXm.
• Application of FXm to fission yeast cells, including those with altered morphology.

Main Results:

• FXm accurately detects subtle differences in fission yeast cell volume, irrespective of cell shape.
• FXm enables high-time-resolution monitoring of dynamic volume changes at the single-cell level.
• The method proves reliable for quantifying cell volume in a small, morphologically diverse model organism.

Conclusions:

• The fluorescence exclusion method (FXm) is a validated technique for precise cell volume measurement in fission yeast.
• FXm overcomes limitations of previous methods, offering dynamic and accurate cell size assessment.
• Integrating FXm with yeast genetics opens new avenues for investigating the fundamental mechanisms of cell growth.