Cells clump after fixation

Priti priti.chugh at gmail.com
Mon Oct 25 10:52:10 EST 2004


I have done porpidium iodide staining for cell cycle analysis several
times but lately i am facing a strange problem with 293t cells. After
fixation when i wash them  they just aggregate and form a mass and
theres no way you can run them on flow cytometer anymore.  Heres an
outline of my experiment. Wshan the cells with PBS, scrape them off
the plate wash again .. resuspend in 70%alcohol while vortexing.. o/n
cold room or -20 or 1hr ... subseqently wash them twice with pbs and
theres an aggregate right there... if not here when they are staining
with PI after the RNase treatment the clump appears.. Please advice.

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