[Histonet] How to get good cross sections of the mouse small intestine

From:"Yu, Jian"

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@upmc.edu



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