Re: 5 Color Analysis

From: RCherv (RCherv@mail-sh.lsumc.edu)
Date: Wed Sep 13 1995 - 21:32:37 EST


     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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