We recently started doing intracellular
staining for cytokines on a FACScan with CellQuest
and are trying to resolve three issues.
1) We see substantial IL-2 and IFN-g positive
cells. Is there a rational, operator-independent
way of setting the markers so that the number of
positives can be identified? Is there software
that will do this?
2) There are a few IL-4 positives and I am not sure that
I want to trust those low percentages? Is there a
better way to detect Th2 cells?
3) Does anybody have experience staining for TNF-beta (or alpha)
or IL-10?
Thanks in advance for any feedback.
Murali Ramanathan
Pharmaceutics