Recycling sections for IHC?

[Perry Mason]

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An extensive IHC workup on a tiny biopsy resulted in it being cut out half way through the panel. Some stains that were done (mainly cytokeratins) were negative. Is it possible to re-stain the slides with a different antibody and what is the procedure?

I did some search and all I found is de-staining H&E slides and re-staining them with immunoperoxidase, which is not exactly the same.

Any push in the right direction would be greatly appreciated.

 

[KCShaw]

I don't recall any personal experience with doing this -- maybe someone else does. But, I'd recommend trying the histotechnology websites for other contacts or documentation, if your own histotechs don't know someone. I believe they have their own listserv. If you're working with a smaller lab or smaller institution, call the nearest large academic center (personally I'd ask the lead histotech) and see if that doesn't put you on the right track.

I have little doubt the problem has come up before and I imagine there's a body of knowledge out there somewhere. The problem you might have is with validity; i.e., there may be decent data with re-staining from X to Y, but not necessarily data specific to what you want to accomplish, like B to C. I don't know. It may turn out to be fairly common and I'm just too far removed.

 

[green mantis]

Just speak to whoever does your IHC. I've done it before. Even negative controls that come back w/ your IHC need to be de-stained 1st. As a precaution for small biopsies, make sure the histotechs don't trim the block before taking sections. It may also be helpful for you to specify the order of the stains. Lastly, if you know you may be ordering stains in batches, you can get unstained slides. That keeps them from continually going back to the block.


----- Antony