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H&E and IMC staining

Posted: Mon Mar 04, 2019 9:16 pm
by mattbliss90
Hello,
Has anyone tried staining the same FFPE slide with haematoxylin and eosin and then an IMC antibody cocktail? If so, does the H&E stain have any negative effect on the IMC antibody binding and the expected end results after running through the Hyperion?

Matt

Re: H&E and IMC staining

Posted: Mon Mar 04, 2019 10:43 pm
by RaulCatena
Hi Matt,

Unfortunately, I did that test and it seems that H/E affects strongly antibody signal as the acid pH of H and basic pH of E strip antibodies from their targets.
This was by performing the staining after antibody incubations, did never tried to pre-stain with H/E, although probably not a good idea either. I´d recommend you to use the I/R trick (where I stands for Iridium, which mimics Hematoxylin, and R stands for Ruthenium, which mimics eosin https://onlinelibrary.wiley.com/doi/abs ... /path.5049) to get a similar image to that of an H/E right off the IMC.