||Murine embryonic stem cells
||Used accutase to detach the monolayer and generate a single cell suspension. Washed with protein-free PBS after centrifugation. Resuspended in annexin V binding buffer (actually, any buffer works here as long as it doesn't contain BSA or something). Generated positive control by treating cells with 0.1% Triton. Pre-diluted Zombie Green 1:10 in PBS and then treated cell suspension 1:100 with the pre-dilution, so ultimately a 1:1000 dilution of ZG was sufficient (therefore, the kit is quite economical). Incubated in the dark, then added annexin V, incubated another 15 mins, centrifuged and resuspended in buffer followed by flow cytometry.
||Gating revealed distinct early apoptotic vs. necrotic populations. Very little background/noise (early apoptotic sample showed fluorescence almost equivalent to the unstained control, for example, as expected).
||Keep everything protected in foil and work in low light, since this is a fluorophore, as usual.