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Flow Cytometry: How to gate Apoptosis? - FSC / SSC Dotplot (May/24/2005 )

I use an Annexin-V / 7-AAD staining for detection of apoptosis. And I wonder how I have to set the gate in the FSC / SSC dotplot for the FITC / 7-AAD diagram...
in the atachmend there is a jpeg with the two regions of cells i usually gate. the upper one is for the living cells and the down one is for the apoptotic and necrotic cells.
Maybe I can expand the gate to the lower left because of the apoptotic bodies beeing released and the apoptotic cell shrinking?
But I think I cannot go too much down, so I detect every apoptotoc body.. or are they too small to be detected whith FACS..??
or shouldn't I gate at all?

I'm happy for every response.



i'm not familiar with your stainning, i stain with PE.
I usually dont have to gate my cells when i'm FACSing because it is quite distinctive.
I would get a distinct dead population in the bottom left corner
while my activated marker cells are anywhere from the middle to the upper right corner of the grid.
there should be a distinct seperation between dead and live cells on the FSC/SCC dot plot.
how many events are you running this thru?