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From:Gayle Callis <>
Date:Fri, 23 Apr 1999 08:22:09 -0600

The murine nasal associated lymphoid tissues and the antigens you want
to stain for will not hold up in NBF fixation/decalcification/paraffin 
sectioning.  We do undecalcified frozen sections, with Instrumedics 
tape transfer.  The markers you stain for do not work well after all those
things, just the nature of the little beasties' cell surface antigens.
I have done whole mouse heads and rat heads fixed with NBF, and decalcified
with 10% formic acid, endpoint tested with xray for both midsaggital
and cross sections of nasal turbinates (when we needed to locate NALT),
with great success, IHC went to the frozens. The hard part to frozens, is
the NALT is located just above the first molar, which is not the easiest thing
to cryosection.

OR there is a way to remove the NALT via a very dicey, microdissection
that works very well.  Learned an interesting thing about NALT, there is a
huge interest in the immunology world about it, but the majority of
literature is in Toxicology journals.  We have a grad student who is an
expert at removing NALT intact, and she has done loads of frozens on the
NALT alone, taking all bone out of the picture with lovely (even double)
IHC staining.  

I also have a publication where they decalcified with EDTA BEFORE fixation,
did frozen sections on this decalcifed bone, fixed the frozens with 
acetone and performed IHC for T and B cells.  It was very elegant and time

Gayle Callis 

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