We digested the brain tissue with Trypsin( 1x) for 15 min (all steps were performed in cold conditions) and suspend the cells by pipetting up and down (3-4 times) to the single-cell suspension. We evaluated the percent of apoptosis, necrosis and live cells using flow cytometry. The result of flow cytometry was found that the percentage of cells in the necrotic phase was enhanced. how can I decrease the necrotic cells?
Jean-Pierre Dangy
What marker did you use for necrosis and apoptosis ? PI and Annexin V ?
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Vahideh Montazeri
yes, the cells were trypsinized and re-suspended in 100 µl binding buffer containing 10 µl Annexin-V-FITC and 5 µl PI .
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