Re: H&E before immuno staining

From:Ross Stapf

We experimented with H/E after immuno staining for a while.  It started when someone accidently counterstained with both Hematoxylin and Eosin.   The Pathologists liked it at first, and asked us to keep doing the mistaken H/E counterstain, but then decided the eosin made it hard to distinguish the DAB at times, and returned to the traditional hematoxylin counterstain.  The brown and orange contrast wasn't always good, but I am sure it depends on what you are trying to see.

We have done immuno's on sections stained H/e because the slides were all that were left to use.  Antigen Retrieval and H2O2 quenching (in our hands) removed much of the h/e stain.

Ross Stapf

Histology Supervisor
Washington Adventist Hospital
Takoma Park MD

>>> Greg Dobbin  02/25/03 07:49AM >>>
Hi Jane,
How about doing the H & E after the immunostain? If your substrate 
is solvent resistant (like DAB) everything should work out fine! I'm 
sure others with more experience than me will also reply.

Date sent:      	Mon, 24 Feb 2003 13:57:59 -0600
From:           	Jane Hosmer 
Subject:        	H&E before immuno staining

> I have a researcher who wants me to stain his slides with H&E before he
> stains with immuno procedure.  After the eosin, should I rinse in any 95%
> alcohols or should I let them air dry immediately after the eosin?   Thanks
> for your help.
> Jane Hosmer, HT,HTL
> CPL Service Center
> UAB Birmingham, Al.

Greg Dobbin
Pathology Lab
Atlantic Veterinary College, U.P.E.I.
550 University Ave.
Charlottetown, P.E.I.
Canada,  C1A 4P3
Phone: (902)566-0744
Fax: (902)566-0851
"A farmer is a person outstanding in their field."

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