Re: [Histonet] querry

From:Rene J Buesa

I never did HIER of frozen sections. Neither on destained pap-smears we used for IHC tests.
We always worked thin, air dried and hydrated FS.
Rene J. Buesa

Vinnie Della Speranza  wrote:
Antigen retrieval is undertaken to "un-do" the masking of tissue antigens that results from formalin (formaldehyde) fixation.

fresh frozen tissues would generally not require this sort of pre-treatment.. unfixed tissues will not be resilient enough to withstand the high heat or harsh effects of extreme pH or proteolysis (depending upon the antigen retrieval method you use), hence you will see the sections come off the slides or look "chewed up"

are you certain that retrieval pre-treatment is necessary in fresh frozen, unfixed tissues, for the antigens you are studying ?

also, in general, thicker sections often have a greater tendency to lift off the slides.

Vinnie Della Speranza
Manager for Anatomic Pathology Services
Medical University of South Carolina
165 Ashley Avenue Suite 309
Charleston, SC 29425
Ph: 843-792-6353
fax: 843-792-8974
>>> "Arvind" 09/27/05 04:23 AM >>>
can any one please tell me how to do antigen retrieval on soft tissue, what 
i do is i took cryosections on slide for human fetal brain tissue age upto 
7 months (prenatal) sections are 40 micron thick, when i do HIER these 
sections are comming off the slides at some points, i use gelatin subbed 
slides for this please guide what to do in detail

thanking in advance

Arvind Singh Pundir

National Brain Research Centre
gurgaon, Haryana, INDIA

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