[Histonet] Re: IHC on frozens

[Johnson, Teri]

In response to this thread:

><snip><
Kimberly:
Your question has a two parts answer:
1- it cannot be done because the sections will peel off, but most importantly
2- it is not necessary since HIER was developed to "undo" the cross linkage produced by the NBF fixation, and the tissues used for?FS are not fixed. ren? J.?

--- On Thu, 7/2/09, Kimberly Tuttle <ktuttle <@t> umm.edu> wrote:


From: Kimberly Tuttle <ktuttle <@t> umm.edu>
Subject: [Histonet] IHC on frozens
To: "histonet" <histonet <@t> lists.utsouthwestern.edu>
Date: Thursday, July 2, 2009, 12:55 PM


Can you do heat retrieval on frozens?

Kimberly C. Tuttle? HT (ASCP)
Pathology Biorepository and Research Core
University of Maryland
Room NBW58, UMMC
22 S. Greene St
Baltimore, MD 21201
(410) 328-5524
(410) 328-5508 fax
Please consider the environment before printing this e-mail.
><snip><

I disagree that you cannot do HIER on frozen sections. We do them all the time. All our samples are fixed in formalin, and then cryoprotected in sucrose prior to freezing them, so providing an antigen retrieval step usually produces good IHC results in frozen sections. We section and dry the slides, then use citrate buffer pH 6.0 in the microwave at 60 degrees C for 10 minutes, cool 10 minutes, then rinse and continue with IHC protocol. Use Plus slides and keep the solution under boiling temperature and you should be fine. For things like brain or bone, you might want to use lower temperatures for longer period of time instead of high temps for less time.

It can even work in fresh-frozen samples which are sectioned and then fixed prior to immunostaining. Some great information on this is included in this article: Yamashita and Okada, Application of Heat-induced Antigen Retrieval to Aldehyde-fixed Fresh Frozen Sections, JHC Vol 53(11): 1421-1432, 2005. A big thanks to Gayle Callis for the heads up on this paper!

Teri Johnson, HT(ASCP)QIHC
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, MO 64110