RE: ask for a protocol for CD3 or CD79a on previously stained slides....

From:Chris van der Loos

Dear Ul Soo,
As those antigens are probably heavily damaged by staining and storing the 
stained slides at room temperature (??), I am afraid CD1a, CD79a and CD3 
IHC staining is going to be extremely difficult, or perhaps impossible. The 
only change you have is trying the tyramide amplification IHC procedure. A 
kit system by Perkin & Elmer (NEL-700) provides the highest amplification 
possibilities for IHC right now. Lots of success.

Chris van der Loos
Dept. of Cardiovascular Pathology
Academical Medical Center
Amsterdam - The Netherlands

Ul Soo wrote:
 >Date: 21 Apr 2003 12:30:23 -0600
 >From: Ul Soo 
 >Subject: ask for a protocol for CD3 or CD79a on previously stained slides....
 >Hi, all!
 >I'm asking for a protocol for applying CD1a, CD3 or CD79a on previously
 >stained slides, especially by Romanoswsky type stains.
 >I'm doing diagnostic cytology in dogs and cats, and I would like to to set up
 >a protocol for previously stained slides by Wright, Giemsa, or May Grunwald
 >Giemsa stains, because, there are increasing tumor cases where we can only
 >obtain a few stained cytologic smears and no mass excised for confirmation!
 >Pet owners avoid surgery frequently for finacial constrains , poor prognosis
 >or lack of information for successful treatment.
 >BUt NO tentative diagnosis without histologic or immunohistochemical
 >confirmation bear any meaning in clinical oncology!!  In fact, I have 
 >to apply some antigens on Wright stained slides and got some successful
 >results, but as for CD antigens such as CD1a, 3, or 79a I have had no idea! I
 >have used antigen retrieval system using citrate buffer,pH6.0 after smear
 >trasfer using Mount Quick to adhesive slides, but the results went
 >unsuccesful.  Any help in any form would be appreciated. If I have ever tried
 >awful things with CD antigens so far, please let me hear No way, or if anyone
 >with experiances immunoCYTOchemistry on Wright stained slides using CD
 >antigens, give me tips for the procedures. Any cautions or advice, or
 >information based on literature would be great help to me.
 >A protocol for applying CD antigens on Wright stained cytologic smears!
 >Thank you in advance.
 >Ul Soo, Choi
 >DVM, graduate student
 >Dept of clinical pathology, Veterinary college
 >Seoul Nat'l Univ, South Korea

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