Dear colleagues, I have a B-D FACScan and routinely do PI staining for cell cycle analysis. I gate on width and area (doublet discrimination). Does this guarantee exclusion of doublets? I can clearly gate out doublets that appear to be 4N DNA content with a large pulse width signal, but I also often see greater than 4N events that do not have a large width pulse. Are these truly >4N cells? The cell lines I use are adherent human tumor and fibroblast cell lines. Any input would be appreciated. Diane Sharp diane.m.sharp@dupontpharma.com
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