[Histonet] RE: H&E staining question - some clarification

[Connolly, Brett M]

I forgot to mention that the H&E was performed in another lab at our facility...most likely on an autostainer.  We just did the IHC studies and I have always used 70% ETOH for tissue storage ... never heard of this staining phenomenon happening before.

Brett

-----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 9:08 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.

Thanks,
Brett

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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