The cell cycle of my MCF10a cells sometimes looks "compressed" with no clearly distinct phases. Other times I get a cell cycle that looks perfectly fine with clear phases and I can´t find a reason why I get such different results. I use DAPI to stain the cells, but I already checked the stain and the protocol and they are alright. So i think it might not be an issue with the staining itself but propably with the cells or the culture conditions or something else?
Has anyone had similar problems or has an idea what the problem could be?
Do you use the same flowcytometer always? Since, detection can vary between different machines and different settings.
Do you specifically have to use DAPI for any reason since, most use PI or 7AAD for this purpose. DAPI could be a problem as machine's LASER power can greatly influence the outcome.
It might help better If you can attach a histogram of good and bad results.
Thanks for your suggestions.
Yes I always use the same flowcytometer with the same settings.
I also tried out staining with PI but the cell cycle looked the same (bad) as with DAPI.
It´s a good point, I´ll attach two histograms with one "good" and one "bad" example. Maype that will clarify what my problem is.
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