Re: 5 Color Analysis
RCherv (RCherv@mail-sh.lsumc.edu)
Wed, 13 Sep 95 20:32:37 cst
Hi...
We're in the process of working out strategies to do 5 color
analysis on a BD FACS Vantage with an Enterprise laser and and HeNe
laser. Thus far, we have settled on FITC, PE, Red613 (excited by
488nm from the Enterprise) and APC (excited by the HeNe). The problem
we have is comming up with a 5th color to use with this laser
combination. We have tried Cascade Blue (in the form of a
streptavidin conjugate used with biotinylated mAbs), but we get a VERY
WEAK signal from this dye (using a 424nm band pass filter for
detection), even with the UV from the Enterprise cranked all the way
up.
Does anyone have suggestions for a 5th color dye we can use with
the other 4? Failing that, does anyone have any suggestions as to
another combination of dyes and filters we can use for 5 color
analysis with this cytometer/laser setup. Any and all suggestions and
comments would be appreciated.
Thanks in advance,
Rob Chervenak
Director, LSU Core Facility for Flow Cytometry
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