[Histonet] Double staining with IF/IHC - chromogenic

[Swartwood, Steven J]

Hello everyone,

I have a co-worker in the research department that would like to know if anyone has ever combined and IF stain with chromogenic IHC staining. His theory for this is that IF shows better nuclear staining from an image analysis stand point. If you have heard of this can you send me literature on it, or if you've done this what is your protocol? It would seem that doing IF first than IHC might cause quenching of the IF, but then again doing chromogenic staining first might cover the IF antigen. I suspect that only an chromogenic cytoplasmic IHC stain can be combined with a nuclear IF marker with IHC first. Any ideas? Again this is specifically from an image analysis stand point.

Steven Swartwood HT(ASCP)
Cedars Sinai Medical Center
steven.swartwood <@t> cshs.org

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