[Histonet] IHC on H&E section

Dana Settembre settembr <@t> umdnj.edu
Wed Feb 11 12:07:35 CST 2004


Patsy,
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" <RossS <@t> BaylorHealth.edu> 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
H/E
stain.  You may have a slight blush of eosin when finished, but it
will
be easy to distinguish from DAB staining in my experience.  I have
recently heard that there are some antibodies that may not be
preserved
after H/E staining, but I have not experienced this personally. 
AE1/AE3
should be fine.

Ross M Stapf
Histopathology Manager
Baylor University Medical Center
3500 Gaston Ave.
Dallas, TX 75246
214-820-2465
214-820-4110 fax
RossS <@t> baylorhealth.edu 



-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu 
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Joanne
Mauger
Sent: Wednesday, February 11, 2004 10:00 AM
To: pruegg <@t> colobio.com; histonet <@t> pathology.swmed.edu 
Cc: ihcrg <@t> yahoogroups.com 
Subject: Re: [Histonet] IHC on H&E section


Patsy,
>From my experience it is possible. I would soak the slide in fresh
>xylene,changing often, until it will lift off without lifting the
tissue.
The H&E stain can be somewhat lightened with acid alcohol, and 95%
alc.
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
takes
days for coverslips to come off. Jo

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


_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet 




_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet 

This e-mail, facsimile, or letter and any files or attachments
transmitted with it contains information that is confidential and
privileged. This information is intended only for the use of the
individual(s) and entity(ies) to whom it is addressed. If you are the
intended recipient, further disclosures are prohibited without proper
authorization. If you are not the intended recipient, any disclosure,
copying, printing, or use of this information is strictly prohibited and
possibly a violation of federal or state law and regulations. If you
have received this information in error, please notify Baylor Health
Care System immediately at 1-866-402-1661 or via e-mail at
privacy <@t> baylorhealth.edu. Baylor Health Care System, its
subsidiaries, and affiliates hereby claim all applicable privileges
related to this information.

_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu 
http://lists.utsouthwestern.edu/mailman/listinfo/histonet




More information about the Histonet mailing list