Re: Mouse Splenocytes

From: Oughton, Julie (julie.oughton@orst.edu)
Date: Mon Apr 23 2001 - 18:02:33 EST


I, too, concur with MAK that one of the populations probably represents dead
cells. However, I just want to add that, in our hands, smooshing a mouse
spleen between the frosted ends of two microscope slides is much gentler
than teasing it apart against a stainless steel mesh. In fact, we routinely
prepare mouse spleen cells using slides, with no more than 2-5% dead cells.

Julie Oughton
Oregon State University




Anjli,
Most likely . . . the population to the left (smaller) is dead.  Confirm
this by adding
PI to your sample, and look for PI staining.  We looked at "smashing"
(assumed, between
slides) vs. "teasing" apart against a wire mesh . . . we found that
viability was far
lower in the smashed organ.
MAK.



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