These AML cell numbers are highest in G1 but move to G2/M with one of the drugs. Another of the drugs appears to induce a minor peak on the lower end of G2/M, and possibly into S-phase. When the two drugs are combined, there is a large peak in this unclear cell cycle phase.
Do you have advice for alternative techniques to clarify this cell cycle stage?
Hello Dawn,
How did you fix your cells? Can you provide one of your said weird profiles? The reason why I am asking is due to the fact most people use ice cold ethanol to fix their cells, which should be added slowly; when added with a higher pace, it ofeten leads to weird profiles.
As an alternative, you can try fixing your cells by heating them up (80ºC should do it) and add Pi afterwards or you can add dyecycle violet, which can stain live cells, thus you don't need to fix them (beware of the lasers and detectors you have available for your cytometer).
Hope it helps!
João Nina
Thank you for your reply. Yes, I am fixing the AML cells by adding EtOH slowly, while continually vortexing to avoid clumping. This peak is not present in all of my flow curves, just when administering particular drugs.
I have attached two example curves from different cell lines. I used to have the cutoff for G2/M at a different location. It is more difficult to determine where the cutoff is when the peak drifts leftward with the addition of a different drug.
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