[Histonet] RE: buffy coat preps

Terri Braud tbraud <@t> holyredeemer.com
Mon Dec 1 12:13:58 CST 2008

I've done lots of buffy coat preps for IHC using cyto-centrifuge preps, though we always found methanol to be the kiss of death to IHC fixation, but don't have a clue as to why. You'd do better to use 95% Alcohol for at least 10 minutes.  Also, by using an alcohol fixative instead of formalin, frequently, much less pretreatment and incubation time is needed.  I used to have about 19 Abs worked out for the Ventana with this method. Sadly, the operative word is "used" to.  I hope this helps.

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Holy Redeemer Hospital and Medical Center
1648 Huntingdon Pike
Meadowbrook, PA 19046
(215) 938-3676 phone
(215) 938-3689 fax

Message: 2
Date: Sun, 30 Nov 2008 23:54:59 -0500
From: John Kiernan <jkiernan <@t> uwo.ca>
Subject: Re: [Histonet] Buffy coat preparation for immuno
Why go to all the trouble of embedding and sectioning a buffy coat specimen? What's wrong with diluting it in saline and then making smears or (better, if you have the equipment) cyto-centrifuge preparations? You'll get lots of slides. They can be fixed in methanol and the WBC stained with any conventional Romanowsky-Giemsa method, or immunohistochemically. You could even fix in formalin, if there's some special reason to do so. (You have to lower the pH of an ordinary blood stain if the fixative was formaldehyde.) 

I hope this message is readable; it is sent as plain text.

John Kiernan
Anatomy, UWO
London, Canada
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