Quantitation of Cell-Cell Fusion of wt Sonicated Samples Using the Multicolor FCM Assay II

The purpose of the experiment is to quantify cell-cell fusion by flow cytometric analysis of a wt X wt. 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. Samples are sonicated before flow cytometry analysis. Results are compared with results obtained with non sonicated samples.
Conclusion:
Cell fusion efficiencies of sonicated and non sonicated wt X wt mating were similar. Each mating sample was analyzed by triplicate, sonicated unstained and compensation controls are included in the experiment