I want to measure cell cycle after a chemotherapy treatment of a B lymphocyte cell line. It may cause cell death.
According to https://www.rockefeller.edu/fcrc/cellsorting/, I should use DPBS (Ca/Mg++ free) and DNAse when working with a sample with a lot of dead cells.
1. a PI stain relies on intercalating in DNA. If we use DNAse, we won't be able to visualize with PI. How do you optimize the buffer for single cell suspension when staining for DNA?
2. Should I use HBSS with Ca/Mg, or DPBS (Ca/Mg++ free), since I am working with cell death in lymphocyte cell line? In addition, an Annexin V kit (https://www.biolegend.com/en-gb/products/fitc-annexin-v-apoptosis-detection-kit-with-pi-8230) requires presence of calcium. Does using a Calcium free- PBS affect my staining?
Thank you!