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We have used AMCA with good results. It is a dimly fluorescent dye, so it
should be used with a "strong" antibody (i.e., CD8 > CD4 > CD3). We used
AMCA in a 3 color setup (with FITC and PE) back before the 488-excitable
red dyes were available. Much nicer than working with a dye laser. :)
An interesting viability dye that we have tried from Molecular Probes is
calcafluor white. In at least one system we noticed the CFW binding to debris
as well as dead cells. CFW is UV-excitable, so you can run 3 colors from
your 488 simultaneously. CFW is *very* bright (turn down your PMTs a few
100V).
(I just re-read your note.) I don't think I'd recommend AMCA with an air-
cooled UV source. We typically used 200 mW UV for AMCA excitation.
/\/\/\_ Eric Van Buren, vanburen%flovax.dnet@rocdec.roc.wayne.edu
\ \ \ Immunology & Microbiology, 313 577 1009
\_^_/ Wayne State University, 313 577 1155 fax
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