Re: FACS on red blood cells

Dave Coder (dcoder@u.washington.edu)
Wed, 19 Nov 1997 10:19:17 -0800

You might want to be sure that the osmolality of all solutions is correct.
The figure of 270 mOs sticks in my head (but that was15 years ago when I
worked with RBCs). A vapor pressure osmometer is convenient way to measure
the osmolality of your suspending media. (People who do electron microscopy
are likely to have one since proper osmotic balance of fixatives is critical
in preserving cellular structure.) Also, fixatives such as formaldehyde
added to normal PBS will increase the osmolality greatly. Red cells are
exquisitely sensitive to changes in osmolality of the medium.

Dave Coder
dcoder@u.washington.edu
-----Original Message-----
From: Monica Nass <mnass@is.dal.ca>
To: cyto-inbox
Date: Tuesday, November 18, 1997 6:50 PM
Subject: FACS on red blood cells

>
>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
>