RE: need help....
Absolutely Vinnie, in fixed tissue rbc's don't lyse. But you can
"decolorize" them as if they were any other hemisiderin containing artifact.
Carson 2nd ed. page 23 option # 2: 70 % ethonal containing 3 mL ammonium
hydroxide for 30 minutes to 3 hours. Wash well with running tap water. Rinse
in 1 % acetic acid and wash again. I have used this technique when using
"bloody" specimens (liver, spleen, bone) for Immuno when the investigator
insisted on DAB peroxidase as the immuno detection color. Works a treat!
Hope that helps, Colleen and any interested others. Donna Montague, UAMS
Center for Orthopaedic Research, Little Rock, AR
From: Vinnie Della Speranza [mailto:email@example.com]
Sent: Thursday, May 10, 2001 2:34 PM
Subject: Re: need help....
I'm skeptical that formalin fixed rbc will lyse. unfixed red cells, as in
the purple top tubes you mentioned, are lysed using an acetic acid solution
which causes the cells to swell and burst. This swelling is the result of
the drawing of water into the cell. whether this same result would occur
with fixed rbc membranes is questionable but someone else may have done this
Vinnie Della Speranza
Manager for Anatomic Pathology Services
Medical University of South Carolina
165 Ashley Avenue
Charleston, SC 29425
ph: (843) 792-6353
fax: (843) 792-8974
>>> Colleen Forster <firstname.lastname@example.org> 05/10/01 02:08PM >>>
It was suggested that I might lyse the RBCS in the formalin specimen.
How would I do this? I cannot remember what
and how much to add to the solution without damaging other cells??/
Also, Lynn, purple top tubes contain EDTA so blood will not coagulate.
However, I doubt there is a fixative as most blood
work is done stat and tubes tossed daily. that is why I wondered if you
could still fix and get anything. Any comments?????
As Always, your help is APPRECIATED !!!!
U of MN, Dept. of Neurology
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