We sort daily using Hoechst 33342 with sperm sexing and we have very good
viability and fertility. As well, we have just completed an exhaustive
experiment with laser power up to 375mW and it showed no decrease in sperm
post thaw motility as compared to 60mW.
We use a 20x160 BSO special design for SX and routinely use 150mW. Sperm
may be more robust than other cells.
My two peaks worth...
Mike
-----Original Message-----
From: ckuszyns@UNMC.EDU [mailto:ckuszyns@UNMC.EDU]
Sent: Thursday, March 15, 2001 2:47 PM
To: cyto-inbox
Subject: Re: DAPI as a viability exclusion dye: Hoechst experience
We have been sorting cells stained with Hoechst dye and followed for 5
days. Cells grow after 5 days of Hoechst. We do not know if there is any
mutagenic effects yet.
Joseph Webster
<J.Webster@centenary.us To: Cytometry Mailing
List
<cytometry@flowcyt.cyto.purdue.edu>
yd.edu.AU> cc:
Subject: Re: DAPI as
a viability
exclusion dye
03/14/2001 05:45 PM
05:34 15/3/2001, Mark Kukuruga wrote:
>Hi all . . .
>Considering the use of DAPI as an exclusion dye, I'm wondering effects of
>about the (albeit brief) UV irradiation on the cells that we sort
>"viably." Should we worry, or is it negligible?
>MAK.
The same question would apply to sorting viable cells stained for DNA
with Hoescht, I've heard of this being done for a long time; anyone
got experience to relate?
After-effects could either result from UV exposure or from the dye
binding direct to DNA; if there are effects, how would you decide
which cause?
Joseph.
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