From: vanburen%flovax.dnet@rocdec.roc.wayne.edu
Date: Mon Oct 31 1994 - 15:03:28 EST
Dr. Fumio Ike had some questions about these dyes. 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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