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I've been spoiled by our Elite (the non-sorting Analyzer cheapo
version) and now I've run into a sticky problem. This user has been
running thymocytes, stained FITC CD4, PE CD8, and Red613 for the
third marker. The Red 613 has a lot of overlap into PE (yes, I'm
painfully aware that this probably isn't "overlap", but poor transfer
efficiency instead), but I can easily subtract both the FITC and the
R613 overlap from the PE channel on the Elite--it allows for subtraction
of 2 signals.
Now they want to sort the high and low Red613 cells, (gated first
on FITC/PE double positives). For that I turned to our 753, and I
can only subtract one signal at a time. There go my nice orthogonal
bivariate plots! We can use an irregular double positive gate, but can
anyone suggest any alternatives? Anyone ever had any luck with this
combination of stains on the older machines? (Anyone willing to loan
me a copy of a successful "core facility improvement" grant
proposal?)
Steve--sorting away, on the thin ice of a new day
(let's see how many nail that reference :-)
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Steve G. Hilliard Cell Analysis Facility
University of Georgia
Surf the FloWeb! <http://www.rserv.uga.edu/cellan/>
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