Contact Erika Fredenburg at HCI Sciences. She and her father may be able to
help you.
Her email is: efredenbu...@hcisciences.com
Toysha N. Mayer, MBA, HT (ASCP)
Instructor, Education Coordinator
Program in Histotechnology
School of Health Professions
MD Anderson Cancer Center
(713) 563-3481
tnma...@mdanderson.org
Message: 1
Date: Tue, 2 Jul 2013 17:24:08 +0000
From: Teri Johnson <tjohn...@gnf.org>
Subject: [Histonet] Re: H&E staining question
To: "histonet@lists.utsouthwestern.edu"
<histonet@lists.utsouthwestern.edu>
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Hi Brett,
I agree with the others, the storage in 70% ETOH is likely not the cause of the
washed out appearance of the staining.
* Did the lab that did the H&E staining run a daily control? How did that
look? Perhaps the tap water they are using went a little funny? Perhaps the
depar xylene needs refreshing? The hematoxylin is used up? If all the other
H&Es done that day look good, then look further.
* Look at the fixative, it is possible to get bad batches of that. It is
also possible to cram a lot of tissue into a small space resulting in
inadequate fixation. Or perhaps it was bloody and not changed to fresh
fixative? Was one sample fixed at room temperature and the other fixed at 4
degrees C? Was the person doing the necropsy the same among animal batches?
* Two things might help get better hematoxylin staining - I recall a
journal article about using antigen retrieval pretreatment to improve H&E
staining on samples that had been stored in fixative for a prolonged period of
time. That might help. Years ago, I also noticed that the hematoxylin
counterstained PAS slides looked better than our H&Es, so we put a bucket of
0.5% periodic acid as an oxidation step before hematoxylin. It needs to be
rinsed well so there is no periodic acid carryover (that'll kill your
hematoxylin!), but that might improve your staining.
If you get this figured out, please let us know the cause and fix.
Teri Johnson
Manager, Histology
GNF - San Diego, CA
858-332-4752
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