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

From:Ul Soo

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 attemted 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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