[Histonet] Processing animal fat

[Liz Chlipala]

The laser treatment should not be an issue, we do that type of work all
of the time.  I would trim to about 2-3 mm.  I did notice that you only
have 2 changes of citrisolv a xylene substitute I have not worked with
that one in particular but if I do work with a xylene substitute we use
three changes and we also have 3 to 4 changes of paraffin depending upon
sample size.  For dermis and skin we have a tendency to process longer
but have never experienced any over processing or "cooked" samples. 

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, Colorado 80308
office (303) 682-3949 
fax (303) 682-9060
www.premierlab.com
 
 
Ship to Address:
1567 Skyway Drive, Unit E
Longmont, Colorado 80504

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Sherwood, Margaret 
Sent: Thursday, March 17, 2011 2:29 PM
To: Grantham, Andrea L - (algranth); HISTONET
Subject: RE: [Histonet] Processing animal fat

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? 

Peggy

-----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
To: HISTONET
Subject: Re: [Histonet] Processing animal fat

Peggy,
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.

Andi


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

> My question is directed specifically to veterinary histologists or
histologists
> 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
them.
> 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
a
> size issue (we trim it if it is greater than 5mm)?  We have gotten
help from
the
> 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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