The lab I do work for has studies about to get underway in rats and monkeys that have blood samples (into EDTA coated tubes) being taken at multiple time points. We have always analyzed fresh blood, but these studies are smaller and it would be better if we could wait until the end to analyze everything instead of wasting time. I am interested in spinning down the blood, removing the plasma, and then freezing the remaining cells for analysis of PBMC subsets. My questions are: 1. We always used whole blood and lysed rbcs after staining, but can you take a blood sample, spin it, take off the plasma, resuspend the pellet, and still get the same values as if you had stained whole blood? 2. Should I just freeze the intact cell pellet after removing the plasma or resuspend the pellet in media and then freeze? 3. If I need to resuspend in media, does it need to contain DMSO or be a freezing media? 4. How long are the cells good for once frozen? Any help would be greatly appreciated. Scott Manetz
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