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Several have discussed the importance of the proper filter selection
(ie. making sure the UV SSC can get to the SSC PMT), but its also
important to be sure that the "side-scatter PMT" is _only_ looking at
SSC. I ran quite a bit of DNA content w/ DAPI about a year ago, and
although we weren't really concerned with SSC, I collected it. I was
confused by the histos for a while, cause I could differentiate
G1-G2M based on SSC. I was using a coverslip as a "dichroic", but I
had no bandpass in front of the SSC PMT, so what I was seeing was DAPI
fluorescence as well as UV SSC! Once I finally realized this, a UV
bandpass in the path cleared up the "mystery" quickly.
This is why I love flow, it keeps you on your toes.
Steve
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Steve G. Hilliard Life's too short
Cell Analysis Facility to drink bad beer!
University of Georgia
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