[Histonet] staining over IHC stained slide

[Curt Tague]

Very insightful response, thank you!



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From: richard cartun via Histonet <histonet at lists.utsouthwestern.edu>
Sent: Tuesday, May 6, 2025 5:00:33 PM
To: histonet at lists.utsouthwestern.edu <histonet at lists.utsouthwestern.edu>; Olson, Jennifer (Carris Lab) <jennifer.olson at centracare.com>
Subject: Re: [Histonet] staining over IHC stained slide

We did this a lot when I was working at Hartford Hospital.  This was especially useful when the lesion or cells of interest in the paraffin block were consumed during microtomy.  The pathologist would select previously tested slides that were negative, or had minimal immunoreactivity, and then reorder the IHC test(s) in the computer, specifying which slide to use for which antibody.  New labels were prepared, and we would then carefully remove the coverslip and then rehydrate to tap water.  There is no need to remove the eosin (which will come out in the alcohol) or hematoxylin (which you want anyway).  No need to quench endogenous peroxidase again, but you will need to perform or repeat the appropriate heat-induced epitope retrieval (HIER) when indicated.  If there is immunoreactivity present in the original slide, you could re-test using a different chromogen to facilitate interpretation.  Obviously, if there is no immunoreactivity in the lesion or cells of interest in the repeat slide, and there is no internal control in the specimen to validate the negative result, the negative test result cannot be reported.
Richard W. Cartun, MS, PhDMorphologic Proteomics, LLCAvon, CT  USA
    On Tuesday, May 6, 2025 at 04:30:18 PM EDT, Olson, Jennifer (Carris Lab) via Histonet <histonet at lists.utsouthwestern.edu> wrote:

 I have a pathologist who is inquiring about staining over and already IHC stained slide. He would like to stain over using a different antibody that has been stained. He doesn't have a specific case or antibody, he is just wondering if there are any histology labs out there that do this?

Any suggestions?

Thank you -
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