[Histonet] Re: rat muscle fixation

[David A. Wright]

Hello Sarah and Histonet

You get wonderful formalin fixation of all rat tissues by transcardial perfusion - and no blood cells. You don't say whether you need a specific muscle and if it needs to be from a specific rat. 

Before you get into the bother of perfusing (& IACUC approval) you might want to contact your local neurology or neurobiology researchers because this is the method of choice for the brain. They will have lots of muscle as a by-product - the induced rigidity of the forelimb muscles (impressive to see the zombie effect) is a standard marker of good perfusion. You could check out some of their muscle to see if it is of a suitable quality. 

I'd ask the veterinarians in your animal facility to suggest someone who is doing this regularly, since this does require institutional approval as a non-survival surgery under deep anesthesia.

-David
==
David A. Wright, Ph.D.
University of Chicago
Section of Neurosurgery, MC3026

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---- Original message ----
Histonet Digest, Vol 103, Issue 22
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Message: 2
Date: Mon, 18 Jun 2012 14:47:46 -0400
From: "Pixley, Sarah (pixleysk)" <PIXLEYSK <@t> UCMAIL.UC.EDU>
Subject: [Histonet] RE: Histonet Digest, Vol 103, Issue 22
To: "histonet <@t> lists.utsouthwestern.edu"
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Dear All:
I am seeking advice from someone who has experience with fixing and paraffin embedding rat muscle tissue. We want to dissect out muscles, fix them, embed in paraffin and then do H&E and immunostaining. Our first attempts, which involved immersion fixation in 4% paraformaldehyde, resulted in either very poor fixation or the tissue got cooked in the tissue processor.  There were parts of the muscle that were hardened and completely disrupted.  We suspect poor fixation, perhaps due to incomplete penetration?  However, if possible, we would like to avoid having to fix via perfusion. Are there any good tricks?   
Please email me off the listserv.
Thanks,
Sarah Pixley
Sarah.pixley <@t> uc.edu