[Histonet] Processing Fat for Paraffin

[Rene J Buesa]

Paula:
For me the times (2 h/each), your dependable instrument and the schedule seem to be OK, but I am not so sure about the Citrisolve because this product (1.4 more expensive than xylene) is made of 91% of water and surfactants + 9% d-limonene and I am not very confident that such a "clearing" agent will be able to allow a proper infiltration on your specimens.
At least for these specimens I would use xylene or a mixture of 2-propanol and mineral oil (attached to another e-mail to you).
René J.

--- On Thu, 4/30/09, Va Paula Sicurello <vapatpxs <@t> yahoo.com> wrote:

From: Va Paula Sicurello <vapatpxs <@t> yahoo.com>
Subject: [Histonet] Processing Fat for Paraffin
To: "HistoNet" <histonet <@t> lists.utsouthwestern.edu>, "MSA BB" <Microscopy <@t> Microscopy.Com>
Date: Thursday, April 30, 2009, 6:22 PM

Hello out there in fat processing land,

I have been given human fat samples and need to embed them in paraffin.  In the
past I've used a VIP processor for this and now I have an Autotechnicon
(vintage dual model) with a timing wheel.

I know I need to process these fatties slowly, my question is--can I use 2
hours per step and have it turn out OK?  I have a timing wheel punched out for 2
hour steps.

My steps would be alcohols: 70, 80, 95 x 2, 100 x 3, Citrisolv x3, paraffin x2
and another paraffin step under vacuum.

Let me know your wise and experienced opinions or protocols.  I don't have
anything else to use except the 43 year old Autotechnicon so don't even
suggest it.  You make me feel bad that I can't get my research foundation to
buy me something new or even newer.  ;-)

Stewing in somebody else's fat (eew!),

Paula  :-)

 
Paula Sicurello
VA Medical Center San Diego
Veterans Medical Research Foundation (VMRF) 
Core Research Imaging Center
3350 La Jolla Village Dr., MC151
San Diego, CA 92161
858-552-8585 x2397


      


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