PKH26 for RBC survival study

From: Jeff Louie (jlouie@hsc.usc.edu)
Date: Thu Mar 15 2001 - 14:42:40 EST 

I have a researcher who would like to utilize PKH26 as a tracking dye
for rabbit rbc's survival studies. I have gotten reasonable separation
of positives and negatives using a "PE" filter setup (555DLP beam
splitter and 580/30 band pass) with 488 excitation on a MoFlo. The
researcher is worried about cell toxicity of the PKH26 and wants to use
as little as possible in his staining procedure. In an attempt to
maximize fluorescence intensity, I've tried to "tune" the dye head laser
(INNOVA 70) on the MoFlo to maximum excitation for PKH26 (551 nm) but
seem to overwhelm the detection PMT with its 580/30 band pass. Any
suggestions or advice on filter setup or use of PKH26 in this scenario
is much appreciated.

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