Quantitation of yeast cell-cell fusion using multicolor flow cytometry

Area of Science:

• Cell Biology
• Microbiology
• Biotechnology

Background:

• Haploid Saccharomyces cerevisiae mating is a key model for studying cell-cell fusion.
• Current gold standard methods for quantifying fusion efficiency are time-consuming and prone to bias.
• There is a need for rapid, standardized, and quantitative methods for assessing yeast mating efficiency.

Purpose of the Study:

• To develop and validate a novel flow cytometry-based approach for quantifying cell-cell fusion efficiency in yeast.
• To provide a faster and more robust alternative to traditional microscopy-based counting methods.
• To enable high-throughput analysis of cell fusion events across multiple yeast strains.

Main Methods:

• Utilized multicolor flow cytometry with Concanavalin A-fluorophore conjugates to stain yeast cell walls.
• Identified mating subpopulations as dual-colored events.
• Discriminated between fused and unfused mating pairs using green fluorescent protein bimolecular complementation.

Main Results:

• The flow cytometry method accurately identified mating pairs and quantified fusion efficiency.
• Results from flow cytometry were indistinguishable from the gold standard microscopy method.
• The developed protocol is simple, reliable, and significantly reduces analysis time.

Conclusions:

• Multicolor flow cytometry provides a fast, robust, and reliable method for quantifying cell-cell fusion in yeast.
• This approach overcomes the limitations of traditional methods, enabling efficient analysis of numerous cell populations.
• The developed technique facilitates high-throughput screening and analysis of yeast mating processes.