[Histonet] use of destained sections in IHC?

Joe Nocito jnocito <@t> satx.rr.com
Wed Feb 23 17:07:37 CST 2011

I wouldn't decolorize the slides. Just remove the coverslip and hydrate to 
water. Any antigen retrieval should take out the Eosin. You're going to 
counterstain in Hematoxylin any way. Years ago (ok, many moons ago) I did an 
informal study. I took 10 antibodies, stained two sets of controls H & E. 
One set, I used 1% acid-alcohol to decolorize the slides, the other set I 
just rehydrated to water. The slides that were placed in acid-alcohol either 
had no staining or a marked decrease in intensity.

----- Original Message ----- 
From: "Walter Benton" <wbenton <@t> cua.md>
To: "Robin Dean" <robin_dean <@t> compbio.com>; 
<histonet <@t> lists.utsouthwestern.edu>
Sent: Wednesday, February 23, 2011 2:00 PM
Subject: RE: [Histonet] use of destained sections in IHC?

It will work and if you are performing antigen retrieval then you won't need 
to decolorize the slide, just rehydrate and perform per you usual protocol.

Walter Benton HT(ASCP)QIHC
Histology Supervisor
Chesapeake Urology Associates
806 Landmark Drive, Suite 126
(All Deliveries to Suite 127)
Glen Burnie, MD 21061
443-471-5850 (Direct)
410-768-5961 (Lab)
410-768-5965 (Fax)
wbenton <@t> cua.md
From: histonet-bounces <@t> lists.utsouthwestern.edu 
[histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Robin Dean 
[robin_dean <@t> compbio.com]
Sent: Wednesday, February 23, 2011 2:43 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] use of destained sections in IHC?

Hi all,

Is it possible to de-stain H&E stained  paraffin sections and re-use them
for IHC staining??? Does anything special have to be done? Will it only work
for some epitopes/stains? Does anyone have suggestions on how to do this? I
am being asked to do this and don't want to waste my time if it won't work.
I would appreciate any suggestions anyone might have

Thank you,


Robin R. Dean, Ph.D.

Senior Scientist & Study Director

Comparative Biosciences, Inc.

786 Lucerne Dr.

Sunnyvale, CA

(408) 738-8060

robin_dean <@t> compbio.com

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