[Histonet] Processing animal fat

Sherwood, Margaret MSHERWOOD <@t> PARTNERS.ORG
Thu Mar 17 15:28:53 CDT 2011

Every station is 1 hour, and our processing program is similar to yours:  we run
the progam overnight, so specimens sit in formalin for a delayed start.  Then
50%, 70% for 30 min. ea.  Then 2x95%, 3x100%, 2xCitriSolv all for 1 hour.  We
end in one change of paraffin. Our specimens are similar in size to yours,
except we keep the thickness below 5mm.

Any thoughts?  Most of our experimental pig skin/fat are laser treated. Could
that be a problem? 


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Grantham, Andrea
L - (algranth)
Sent: Thursday, March 17, 2011 4:10 PM
Subject: Re: [Histonet] Processing animal fat

Is your whole program one hour or is each station one hour?

We had a project here a few years ago that we called "the bacon project" because
we had whole chunks of pig skin with an implant and lots of fat in between the
layers. It was fixed for a couple days in 10% NBF and then processed with one
hour for each station starting in 70% ETOH and ending in 4 paraffins on a VIP.
Turned out beautiful. Did trichromes and they were stunning - red, white and
blue! The sections were big - about 2.5 cm square - took almost all of the space
in the cassette.


On Mar 17, 2011, at 12:57 PM, Sherwood, Margaret wrote:

> My question is directed specifically to veterinary histologists or
> who also do a fair amount of animal processing.  We are having a terrible time
> processing pig fat.  We had problems previously, but thought we had solved
> This latest project (pig skin with a lot of fat attached) came out awful.  The
> fat was not adequately processed:  couldn't section it, it just crumbled.  In
> the block, it appears white and crumbly.  The funny thing is some blocks came
> out all right, but most didn't.
> PLEASE help!  Let me know how you process your animal fat (sp. Pig)! Is there
> size issue (we trim it if it is greater than 5mm)?  We have gotten help from
> histonet before and instituted these suggestions (i.e. let sit in formalin for
> 48 hours; put in cassettes in a 60 degree oven for 1-2 hours and then cool to
> room temp. Then process using a 1 hour program).
> Any help would be appreciated.
> Thanks!
> Peggy
> Peggy Sherwood
> Lab Associate, Photopathology
> Wellman Center for Photomedicine (EDR 214)
> Massachusetts General Hospital
> 55 Fruit Street
> Boston, MA 02114-2696
> 617-724-4839 (voice mail)
> 617-726-6983 (lab)
> 617-726-1206 (fax)
> msherwood <@t> partners.org 
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