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 --------------------------------------------------------------- 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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