re: PI gating on FL3
KUKURUGA@medmail.med.umich.edu
Thu, 31 Jul 1997 14:19:16 -0400 (EDT)
Assuming you're talking about a three-color analyzer . . . looking at PI
in the "FL3" (>650nm) channel will work OKAY. Keep in mind that you will
see PI in "FL2" (~575BP) as well, but it's not a problem because you're
excluding the cells that are PI positive, so they don't factor in. There
can be an increase in noise to the PE channel, though, so it's probably good
to compensate for it with single color controls.
Anyway . . . yes, your approach is sound. I've even done the same looking at
the PI signal in the PE channel, and eliminating the very brightest cells
as the "dead" (PI+) cells. This can work on a machine with only two
fluorescence PMTs.
Alternatively, one can use 7-AAD as an exclusiond dye, and not worry so
much about crossover, since the 7-AAD emits well into your "FL3" parameter's
band range.
MAK.
Mark A. KuKuruga
University of Michigan Flow Cytometry
kukuruga@medmail.med.umich.edu