accepted fixation protocols

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From:Judy Trogadis <> (histonet)
Date:Mon, 15 Mar 1999 12:10:32 -0500 (EST)

Dear Histonetters:

I am trying to stain human lens sections, first with fluorescent markers
for identifying fiber cells and nuclei, then with other antibodies to
study cataract formation.

The tissue is human archival eyes, formalin fixed and paraffin embedded. 
So far, the results include mostly autofluorescence. My question is the 
following: Is it the accepted convention that paraffin sections are used
for immunohistochemistry and cryosections for immunofluorescence? That is
my impression from doing a literature search. The other explanation may
be that routine clinical labs use paraffin embedding and immunohistochem.
because it is more common (?faster, ?easier) and immunofluorescence is
more of a research tool, although both disciplines use cryosections.
Any opinions?

Unfortunately, the most critical antibody I want to use ultimately so far
has only worked with immunofluorescence so I will have to try quenching
the autofluorescence. How realistic is that with lens, especially 
cataractous lens?

If anyone knows tha answers to these questions, it is this group, so
thank you in advance.


Judy Trogadis
Eye Research Institute and
University of Toronto
Toronto Hospital, Western Div.
399 Bathurst  St.
Toronto, Canada M5T 2S8

phone: 416-603-5088
Fax:   416-603-5126

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