 |
 |
 |
 |
We are trying to work up a 3 or 4-color set up, that allows some
intracellular staining, with some surface staining, and excludes cells
that were "dead" or membrane permeant prior to the fixation step. The
main problem is selection of the stain for "dead cells". Candidates are
PI or TOPRO-3, to indicate cells that are membrane permeant and label
their DNA before all other labels and fixation.
The cells are freshly isolated murine lymphoid cells (populations of B
and T cells present)
Setup: 3 color potential on 488 (FACscan)
4 color with 488 plus 630 (HeNe) or 488 plus UV - (FACStar plus)
Suggestions for other "dead" cell stains are welcome, including non-DNA.
Please tell us about washing requirements etc. I tried a simple stain
with PI previous, but noticed some loss of signal, following all the
fixation and washing. Is there a more "permeant" DNA stain to mark
"dead" cells?
Denis Snider PhD, FCACB
Associate Professor
Dept. of Pathology
McMaster University
Tel. 905-528-2010, FAX 905-521-2613
|
|
|
|