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From: Murali Ramanathan (murali@ACSU.Buffalo.EDU)
Date: Fri Jan 17 1997 - 10:35:03 EST

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Hi Everybody,

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

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