Epithelial cells and apoptosis

From: Steve G. Hilliard (steve@habanero.cb.uga.edu)
Date: Wed Feb 04 1998 - 08:56:45 EST


Greetings from soggy Georgia!

A quick question--does anyone have any special tips or precautions for
preparing epithelial cells (a kidney cell line) for any type of flow
cytometric apoptosis assay?  One of our labs has had good success with
avian bursal lymphs using PI, tunel, MC540 and CMXRos, but a new user is
having no success applying these protocols to the aforementioned
epithelial cells.  Scatter gate usually shows a very diffuse population of
cells and a significant percentage that I would consider debris, but
we get no tunel labelling, and using PI we get 50% with extremely low 
fluorescence (which back-gates to the "debris") and a mere bump
instead of a sharply defined diploid peak.  I'd love to hear from
anyone who has any experience with epithelial cells.

Thanks,
Steve
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Steve G. Hilliard        steve@habanero.cb.uga.edu        
If you're a runner you should be dead. 
http://storm.cadcam.iupui.edu/drs/drs.html
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