Quantitation of Cell-Cell Fusion Using the Multicolor FCM Assay III

The purpose of the experiment is to quantify cell-cell fusion by flow cytometric analysis of a wt X wt and a prm1∆ X prm1∆ yeast mating. MATa and MATalpha strains are distinguished by staining each strain with ConcanavalinA- Alexa Fluor 647 (ConA-647) and with ConcanavalinA- Tetramethylrhodamine (ConA-Tet), respectively. Mating pairs are revealed as two-colored entities. Cytoplasmic mixing is measured with a GFP bi-molecular fluorescence complementation assay. Cell fusion efficiency is calculated as the percentage of fused mating pairs over the total number of pairs.
Conclusion:
Cell fusion efficiencies of a wt X wt mating and a prm1∆ X prm1∆ mating were quantified. Each mating sample was analyzed by triplicate, unstained and compensation controls are included in the experiment.