Re: CRBC/gluteraldehyde

jphillip@mednet.med.miami.edu
Thu, 06 Apr 95 16:43:15 EST

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Joseph,
It is pretty hard to mess up with CRBC's. The way that I do it is to
first find a chicken. Working in a 50ml tube, put the blood in some
alsevers anticoagulant. When time allows, spin and wash 2x. Save the
red cells only. Dump in some 8% gluteraldehyde (one of those little
glass vials, aprox 4 or 5 mls) and put the tube on a rotator (rotating
mixing plate) in a cold room for 2 days. Wash the cells with PBS 3 or
4 times and bring them up in a 2% gluteraldehyde solution and leave
them in this. They will stay good on the bench top for approximately
4 YEARS. Two things to watch out for. The cells keep gaining
fluorescence for the first couple of months, and be sure to wash the
sample line well before running a sort sample. You don't want to fix
the sort cells.
Jim

______________________________ Reply Separator _________________________________
Subject: CRBC/gluteraldehyde
Author: J.Webster@centenary.usyd.edu.AU (Joseph Webster) at smtpmed
Date: 4/6/95 2:30 PM

Would someone please send me the recipe for gluteraldehyde fixation of
chicken erythrocytes?

I have numerous references, but they are all in books or journals that I
can't quickly lay hands on. (Do others find holes in the bookshelves?)

Many Thanks,
Joseph.

Joseph Webster (O.I.C. Flow Cytometry & Computing) ===
Centenary Institute of cancer Medicine & Cell Biology / \
Locked Bag No.6 || o o ||
Newtown, NSW 2042 || | ||
AUSTRALIA. | \___/ |
Ph: 61-2-565-6110 \ /
Fax: 61-2-565-6101 |||||
E-Mail: J.Webster@centenary.usyd.edu.au

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