[Histonet] How to get good cross sections of the mouse small
intestine
Yu, Jian
YuJ2 <@t> upmc.edu
Mon Jan 23 15:11:28 CST 2006
Dear all,
I am a new histoneter. I am just starting to do some mouse small
intestine histopathology and wondering whether anyone can provide a
method (protocol) for how to fix and embed mouse small intestine to
generate good cross (paraffin) sections with as many intact villi and
crypts possible for H&E and IHC Particularly, I need to evaluate at
least 5-10 different area of the small intestine to score the
regeneration of crypts and the extend of apoptosis at various time point
after irradiation.
I have used neutral buffered formalin as fixative and manually bundle
four pieces of 0.5 cm-long tubes together before embedding at a clinical
pathology lab. The sections looked OK, but many appear to have a lot of
floating villi in the lumen and limited number of crypts extending into
to the villi connected to them. Since we can not always get our
tissues processed after O/N fixation, we sometimes keep the tissues in
NBF for several weeks. Is this too long, or should we transfer them to
70% ethanol?
Look forward to your expert suggestions.
With many thanks,
********************************************************
Jian Yu, Ph. D.
University of Pittsburgh Cancer Institute
Hillman Cancer Center Research Pavilion
Office suite 2.26h, Laboratory 2.43
5117 Centre Avenue, Pittsburgh, PA 15213
Phone : 412-623-7786, (Lab) 412-623-3255
Fax: 412-623-7778
Email: yuj2 <@t> upmc.edu
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