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 <[email protected]> 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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