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

[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,

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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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