RE: [Histonet] IHC on H&E section

From:Dana Settembre

Soak in the same reagent that was used before coverslipping.  If a
xylene substitute was used, soak in that.  We use xylene, so we soak in
xylene to remove coverslip.  Slides coverslipped yesterday come off
within an hour. If coverslipped two years ago, it could take days at
room temp.
I agree with Ross Stapf regarding destaining.  It is not really
necessary.  I use a citra pretreatment in the microwave when running
AE1/AE3 and the color left behind is a mild blush.  I run immunos on H&E
slides often and it  works.

Dana Settembre
University Hospital - UMDNJ, Newark, NJ

>>> "Stapf, Ross"  2/11/2004 8:24:44 AM >>>

In my experience destaining is not necessary if you are doing a heat
epitope retrieval.  The retrieval will remove most if not all of the
stain.  You may have a slight blush of eosin when finished, but it
be easy to distinguish from DAB staining in my experience.  I have
recently heard that there are some antibodies that may not be
after H/E staining, but I have not experienced this personally. 
should be fine.

Ross M Stapf
Histopathology Manager
Baylor University Medical Center
3500 Gaston Ave.
Dallas, TX 75246
214-820-4110 fax 

-----Original Message-----
[] On Behalf Of Joanne
Sent: Wednesday, February 11, 2004 10:00 AM
Subject: Re: [Histonet] IHC on H&E section

>From my experience it is possible. I would soak the slide in fresh
>xylene,changing often, until it will lift off without lifting the
The H&E stain can be somewhat lightened with acid alcohol, and 95%
The destaining is not really necessary. Depending on the
antigen,hopefully it is preserved, the IP staining should work as
usual,with a little dirty H&E background. Good luck. Sometimes it
days for coverslips to come off. Jo

>>> "Patsy Ruegg"  02/11/04 10:36AM >>>
I know this has come up before but don't remember the details.  I
like to do IHC for AE1/AE3 on a slide from a study set that has been
stained and coverslipped for some time now.  Please advise.  Is there
preferred method for removing the coverslip and maintaining the
integrety? Destaining the H&E?  Doing the IHC stain on this section?

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