Hello Adrian,
I have used the UV channel quite a bit for live/dead cell
discrimination, using Ho33258 and 33342. It works
quite well, and compares nicely with PI.
DAPI works well also. The Ho33258 works
better than the 33342 on ice for this application. I've done this
on a Partec, MoFlo, and Vantage systems with similar results.
I have not done this on the LSR, but I see no reason why it
wouldn't work there also.
Check out the following:
Lopez, P.A., Ottenberg, M.O. Rapid cellular viability quantification using
an arc-lamp flow cytometer.
Cytometry 1998; Suppl. 9:143
It's essentially the same Ho staining protocol as here:
Hardin,J.A., Sherr,D.H., DeMaria,M., Lopez,P.A. A simple fluorescence
method for surface antigen phenotyping of lymphocytes undergoing DNA
fragmentation. J. Immun. Methods 1992; 154:99-107
Peter Lopez
The Aaron Diamond AIDS Research Center
212.448.5188 (office)
212.448.5159 (fax)
212.448.5190 or 5110 (lab)
Adrian Smith
<A.Smith@centenary.us To: Cytometry Mailing List
yd.edu.AU> <cytometry@flowcyt.cyto.purdue.edu>
cc:
02/22/2001 11:15 PM Subject: DAPI vs PI for dead cell
discrimination
Hi all,
We are currently evaluating a B-D LSR in order to decide
whether it meets our requirements. One possible configuration we are
considering would use DAPI excited by the UV laser (325nm) for
live-dead discrimination in place of PI. I know we don't need to
devote a channel to PI but some users would prefer to have a separate
live/dead channel and in most experiment we wouldn't be using the UV
laser for anything else.
My question is has anyone used DAPI for live-dead discrimination? How
does it compare to PI? Are there any particular issues we need to be
aware of?
Thanks,
Adrian Smith
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