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

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. Conclusion: Cell fusion efficiency of a wt X wt non sonicated mating was similar to sonicated samples of experiment titled Quantitation of Cell-Cell Fusion of wt Sonicated Samples Using the Multicolor FCM Assay I. Unstained and compensation controls were included in the experiment, matings were analyzed by triplicate.