Hi all- I have never responded to the flow list before so I have no idea if this will even make it there, but here goes.... I've been labeling cells, mostly human lymphocytes, with CSFE for nearly a year. These have been used in mixed lymphocytes culture, other proliferation assays and as targets for CTL assays. One thing I've learned through trial and error that hasn't been mentioned yet is letting cells "rest" in 20% FCS/RPMI in an incubator for 2-4 hours after thawing. This seems to yield better staining and more functional cells. I also have found if I turn down the Fl-2 PMT substantially, I only need about 30% compensation between Fl-1 and Fl-2. I tend to set my primary CSFE (un-proliferated) peak around 200 on a 256 channel scale. Mary Gallagher Research Associate Puget Sound Blood Center Seattle, WA 98104 -- End --
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