I've done lots of buffy coat preps for IHC using cyto-centrifuge preps, though we always found methanol to be=20the 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=20hope this helps.

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


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Message: 2
Date: Sun, 30 Nov 2008 23:54:59 -0500
From: John Kiernan 
Subject: Re: [Histonet] Buffy coat preparation for immuno
T
Why go to all the trouble of embedding and sectioning=20a buffy coat specimen? What's wrong with diluting it=20in saline and then making smears or (better, if you=20have the equipment) cyto-centrifuge preparations? You'll get lots of slides. They can be fixed in methanol and=20the 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=20have to lower the pH of an ordinary blood stain if=20the fixative was formaldehyde.) 

I hope this message is readable; it is sent as plain=20text.

John Kiernan
Anatomy, UWO
London, Canada
= = =

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