Dear Fellow Flowers: I'd like to ask my more learned flowlosopher colleagues a question on gating for the purpose of quantitating sub G1 content for apoptosis studies. I'm hoping Powerpoint illustrations can be attached to these mass mailings. We all know that we should gate out doublets and triplets in order to restrict a cell cycle analysis to single cells and one of the ways of doing this is to plot Area vs Width in order to draw a region. The Powerpoint attachment is a dot plot of such an experiment. Notice the long trail of sub G1 events that are big, fat and wide. Should these events be gated out as in the first histogram or left in as in the second histogram? It seems to me that if they are two or more sub G1 events are stuck together they're still apoptotic and should be left in the gate otherwise it's cheating the apoptotic population of its due representation. However, I've never seen a discussion of this. So, if some of you have any insight into this, I'd like to hear it. Sincerely, Dan Rosson Dan Rosson Ph.D. Lankenau Institute of Medical Research 100 Lancaster Ave. Wynnewood, PA 19096 www.limr.org.
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