Hi Mark,
That's a good question, and although I've never heard
people complain about viability after a sort with this technique,
I never did the actual post-sort viability study. My experience is
using Hoechst, not DAPI, but in either case I wouldn't be worried.
The sperm sorting folks have looked into this. They do UV excited
Hoechst 33342 sorting daily with good results.
The live/dead app gives a bright staining so you don't need much
UV power. I've had good resolution using a sick argon 90-5 which
only put out 30mw of UV . I bet you could even use half that power.
The viable cells are negative, so they don't really pick up much dye, if
that is the concern.
Peter Lopez
The Aaron Diamond AIDS Research Center
212.448.5188 (office)
212.448.5159 (fax)
212.448.5190 or 5110 (lab)
"Mark
Kukuruga" To: Cytometry Mailing List
<kukuru@med.u <cytometry@flowcyt.cyto.purdue.edu>
mich.edu> cc:
Subject: DAPI as a viability exclusion
03/14/2001 dye
01:34 PM
Hi all . . .
Considering the use of DAPI as an exclusion dye, I'm wondering about the
effects of
(albeit brief) UV irradiation on the cells that we sort "viably." Should
we worry,
or is it negligible?
MAK.
--
Mark A. KuKuruga, Managing Director
University of Michigan Flow Core
7416 CCGC 0946
(734) 647-3216, fax (734) 936-7376
kukuru@umich.edu
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