I have done FACS analysis on human red cells. I usually wash them and do all the staining in RPMI w/ 1% BSA then I fix them w/ 1% paraformaldehyde. Prior to the analysis I bring them up in PBS and I have noticed than within a few minutes they lyse but this is long enough to do the analysis. I do not know if some of these techniques could be applied to porcine cells. Let me know. Jose A. Stoute, MD _______________________ Reply Separator _______________________ Subject: FACS on red blood cells Author: Monica Nass <mnass@is.dal.ca> at wrsmtp-ccmail Date: 11/17/97 1:43 PM Hi everyone. Newcomer here. Please help! I've been trying to perform FACS on porcine red blood cells with no success. I can't get the cells to survive the labelling process. When BSA is present, the cells get all crumpled up. I've tried storing the cells in Alsever's (a citrate dextrose solution) prior to use, but when I wash off this Alsever's several times in PBS and aliquot the cells into small numbers (1 million per tube), I can no longer get the cells to spin down! Maybe I need something else for storing the cells. I've tried plain PBS, but the cells again crumpled up. Any advice would be greatly appreciated. Monica Nass Research Associate Dalhousie University Halifax, Nova Scotia
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