[Histonet] RE: H&E staining question

Bea DeBrosse-Serra BDeBrosse-Serra <@t> isisph.com
Tue Jul 2 10:18:30 CDT 2013

Hi Brett, 

That the H&E staining looks faded after storage in 70% seems very unlikely. We do this with rat and mouse tissues all the time. Did the other lab use a different Hematoxylin, Eosin, or used a different staining protocol on the autostainer?


Beatrice DeBrosse-Serra HT(ASCP)QIHC
Isis Pharmaceuticals
Antisense Drug Discovery
2855 Gazelle Ct.
Carlsbad, CA 92010

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Connolly, Brett M
Sent: Tuesday, July 02, 2013 6:08 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] H&E staining question

Histonetters -

We had some rat tissues that were removed at necropsy and immediately fixed in formalin for 48 hrs.  Since the tissues could not be processed after the 48 hr fixation they were transferred to 70% ETOH for about a week, then processed and stained with H&E.

The H&E staining looks faded/washed out when compared to previous rat tissue that was fixed for 48 hr and then processed and stained without the extended time in 70% ETOH. The reason for limiting the fixation to 48 hr in the more recent study was that we were doing some IHC stains and wanted to keep the same fixation time as the previous study.

I have extra unstained slides left over - any suggestions on how to get some better, crisper H&E results?  Has anyone observed the washed out H&E appearance from tissues stored in 70% ETOH.


Brett M. Connolly, Ph.D.
Principal Scientist, Imaging Dept.
Merck & Co., Inc.
PO Box 4, WP-44K
West Point, PA 19486
brett_connolly <@t> merck.com
T- 215-652-2501
F- 215-993-6803

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