[Histonet] Mouse pups and processing

From:"Amos Brooks"

     We were having a similar problem with one of our researcher's P0 mice.
I don't know how they sacrificed them, but they too dod not process at all.
They were squishy!
     I reccommended they either bisect or at least make a good midline
incision prior to fixation. It still isn't perfect (is it ever?), but it
seems to be a bit better. This allows better solution penetration. Give it a
whirl and let us know how it goes.


Message: 13
Date: Fri, 08 Feb 2008 15:52:34 -0500
From: Pamela Marcum 
Subject: [Histonet] Mouse pups and processing
To: Histonet@lists.utsouthwestern.edu
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I have been struggling with this for a while and need some help.  We
currently have a project with 0 day mouse pups that are allowed to be
born normally and then sacrificed with CO2.  We had several groups
earlier that were sacrificed a different way and they processed and
sectioned beautifully.

These don't seem to fix well, dehydrate, clear or infiltrate worth a
darn.  Since this is my first time using CO2 for sacrifice I need to
find out if it causes a problem or if I am just losing it.  I have
not ever had this problem before and even re-processing does not
help.  I know they are not infiltrating as they are floating in
paraffin at the end if I remove them from the processor to a vat of
paraffin.  They will not sink.  The pups are 1.2 to 1.3 grams each.

If you can suggest a better way to sacrifice them please let me
know.  Killing the mother and perfusing her is not an option as these
are not our mice.  They are being given as favor so I am limited to
some extent.

This was an overnight process with slightly altered alcohols to 45
minutes each, 1 xylene at 30 minutes and 2 Shandon Xylene Substitues
at 10 hour each to 4 paraffins at 45 X2 and 1 hour X2.

Best Regards,

Pamela A Marcum
Manager, Histology Special Procedures
University of Pennsylvania
School of Veterinary Medicine
R.S. Reynolds Jr.  CORL
New Bolton Center
382 West Street Road
Kennett Square, PA 19348

Phone - 610-925-6278
Fax     - 610-925-8120
E-mail - pmarcum@vet.upenn.edu
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