Re: IHC question

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From:Lee & Peggy Wenk <>
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We use a 50:50 mixture of acetone and xylene. Comes off in less
than an hour, sometimes as little as 10 minutes.

Histology stains look fine with it. I don't have direct
experience about IHC staining after removing the film
coverslips. I have done it on slides that hand been
coverslipped with glass.

But logic seems to indicate that, since the tissue section 
has already been through hematoxylin (with all its chemicals), 
eosin (with all its chemicals), xylene, dehydrant (usually 
alcohol), a bluer, and possibly an acid rinse, that the
tissue antigens have already been subjected to a lot more
chemicals that are possibly worse. If all these chemicals
don't destroy the antigen (and most do seem to survive on 
H&E stained slides that have been coverslipped with glass), 
I don't see why putting the slide in another change of 
dehydrant (acetone) and xylene would hurt the antigen.

My hunch, which may be wrong.

Peggy A. Wenk, HTL(ASCP)
William Beaumont Hospital
Royal Oak, MI 48073

> Dear Histonetters,
> For those of you who do IHC stains and get slides that have been previously stained e.g. H&E, how do you get the coverslip film off the slides?  I know there is a quick method by placing the slides in acetone, but does this affect the antigenicity of those slides?
> Right now I soak them in Xylene, but it takes days to get the film unglued.  Glass coverslips come off rather quickly in xylene, it's just the film coverslip that takes forever.
> Thanks for whatever info you can share.
> Ann Maruska
> Fairview-University Med Ctr
> Mpls.  MN  55454

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