effect of viability dyes on single cell culture of human lymphocytes

Guy Hermans Guy.Hermans at ablynx.com
Fri Jun 22 04:12:37 EDT 2007


Hi J.,

Since the dyes only enter dead cells, short-term toxicity probably isn't
the problem. If it were, all your cells would light up before the end of
the sort ;-)

A single sorted droplet sorted in cell culture medium in a normal
96-well plate-type volume gets diluted extremely far - and since most of
that volume is sheath, not sample, fluid anyway there shouldn't end up
much free dye in the destination well/tube. This assumes they don't
stick to the cell membrane of something, of course.

That being said, I routinely use TOPRO3 and that doesn't seem to inhibit
outgrowth significantly. I never really compared with and without dye
head-to-head, since I always wanted to exclude dead cells anyway.

Guy

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-----Original Message-----
From: James Crowe [mailto:james.crowe at Vanderbilt.Edu] 
Sent: Wednesday, June 20, 2007 9:45 PM
To: cyto-inbox
Subject: effect of viability dyes on single cell culture of human
lymphocytes


I am interested in whether anyone has or has seen data (ie not just  
opinion) on:

Whether addition of one or multiple viability dyes to human  
lymphocytes being sorted as single cells for later culture and	
proliferation affects the ability to proliferate those single cells.

Certainly, sorting more live cells and excluding dead cells is	
desirable, but also it is generally thought PI and some other dyes  
exert a toxic effect on prolonged exposure.

J. Crowe



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