From: Bethany Mundy (mundy-2@medctr.osu.edu)
Date: Thu Oct 02 2003 - 11:52:50 EST
I wanted to clarify my earlier question regarding 7AAD staining with fixed cells. I was referring to staining the cells with 7AAD while fresh, washing, and then fixing them. The fixation step obviously disrupts the membrane, which is why a membrane dye can not be used after the cells have been preserved. Basically, it looks like the dead or apoptotic cells stain brightly from that initial labeling step, but if any 7AAD dye is left in the tube after the wash, when the cells are lysed for fixation, it lightly stains the live cells too. This can lead to three peaks: dead, live, and debris. I was wondering if anyone else noticed this situation. Thanks again, Bethany Bethany L. Mundy Research Associate Department of Pathology College of Medicine and Public Health The Ohio State University mundy-2@medctr.osu.edu
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