[Histonet] Breast Processing Protocol

[Edmondson David (RBV) NHS Christie Tr]

Hi all,
We have been using Carnoy's fixative as an intermediate / degreasing stage.   Cassettes in a pot of it for a few hours,  it would be better on an old processing machine "Dunking" but space does not allow.   We rinse the cassettes before processing so we do not get the full benefit of any dehydration that may have occured.
Dave,
Christie Hospital
Manchester UK

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Laurie
Reilly
Sent: 18 August 2005 00:33
To: DDDeltour <@t> mar.med.navy.mil; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Breast Processing Protocol


Douglas and fellow histonetters,

The major problem with processing fatty tissues, assuming that they are
fixed properly, is that Ethanol is not a good solvent for fat and therefore
connot penetrate the tissues completely, so the tissues are inadequately
dehydrated.
We have had some success with lipomas by adding a "degreasing" step of
xylene into the processing schedule.
70% ethanol
80% ethanol
90% ethanol
95% ethanol
Absolute ethanol
Xylene
Absolute ethanol
Xylene
Xylene
Paraffin
Paraffin
Paraffin
The first Absolute ethanol will dehydrate the tissue to some extent. The
next Xylene step will remove most of the fat and then the second Absolute
ethanol can complete the dehydration.

A compromise situation that we use routinely is to have
Absolute ethanol
50:50 Absolute ethanol:Xylene
Xylene
This is not quite as effective but it is less disruptive to the normal
schedule and handles moderately fatty tissues.

            Regards from Townsville, Australia.
                      Laurie.


At 09:58 AM 17/08/2005 -0400, DDDeltour <@t> mar.med.navy.mil wrote:
>I am looking for a program for processing fatty breast tissue. We currently
>let the breast sit in cassettes in Pen-Fix for an extra day before
>processing. We then process the first Fixative in Pen-Fix for one hour then
>in 10% Formalin for the second hour (I do not like this but that is they way
>it was done when I got here). The next steps are.. 95% one hour X3, 100% one
>hour X3, Xylene 1 hour X2. Paraffin for one hour X3. I am considering
>replacing one of the 95% with another fixative. What programs do you use for
>this? They don't like to hear Sh in Sh out:-). Thank you!
>
>
>
>DOUGLAS D. DELTOUR
>HISTOLOGY TECHNICIAN
>NMC PORTSMOUTH VA
>
>
>
>_______________________________________________
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>Histonet <@t> lists.utsouthwestern.edu
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Mr.Laurie Reilly                                              Ph 07 4781 4468
School of Veterinary & Biomedical Science      Fax  07 4779  1526
James Cook University
Townsville  Qld. 
4811                                      laurie.reilly <@t> jcu.edu.au 

Australia.


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