Re: FACS on red blood cells

From: LTC Jose Stoute (stoute@wrsmtp-ccmail.army.mil)
Date: Fri Nov 28 1997 - 06:08:02 EST


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

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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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