From: Stetler-Stevenson, Maryalice (NIH/NCI) (stetler@mail.nih.gov)
Date: Mon Nov 18 2002 - 18:01:52 EST
I have a colleague who does some flow on the side but doesn't know much about the technique. He asked me about a strange artifact he observed in staining with a FITC antibody and PI (transfected cells- looking at expression of a protein and cell cycle). I thought it was a compensation problem and I suggested he try compensation to get data he could analyze by a cell cycle program. The next day he told me he decided it was FRET and therefore it couldn't be compensated (not that he tried it of course- he just decided that the protein was in the nucleus and close to the DNA). I think he has much to do to prove FRET but since he has published bad flow data before in non-flow journals I don't imagine he will have any incentive to do so. I wondered- could you compensate out FRET to be able to get cell cycle plots that you could analyze? Maryalice Stetler-Stevenson Chief, Flow Cytometry Unit Laboratory of Pathology, NCI, NIH Sometimes you're the windshield, sometimes you're the bug.
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