[Histonet] RE: H&E staining question

Tony Henwood (SCHN) tony.henwood <@t> health.nsw.gov.au
Tue Jul 2 19:16:51 CDT 2013

You could try 10 minutes treatment with periodic acid prior to H&E staining (Luna's technique I think)

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager & Senior Scientist 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Connolly, Brett M
Sent: Tuesday, 2 July 2013 11:08 PM
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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