[Histonet] RE: Oil Red O staining question

[Manfre, Philip]

We pretty much only used formalin-fixed, frozen tissue for ORO.  Just don't process them - the alcohol will dissolve the lipid out.

Phil.

Philip Manfre, B.A., HT (ASCP)
Associate Principal Scientist
Merck Research Laboratories
WP45-251
PO Box 4
West Point, PA 19486

215-652-9750
215-993-0383 (fax)
philip_manfre <@t> merck.com





-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala
Sent: Thursday, March 20, 2014 12:30 PM
To: David Burk; histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: Oil Red O staining question

David

That does work, we have done it here on mouse muscle and liver,  but there are some problems to fixed and then frozen tissue.  The tissue  on occasion does not stay on the slide as well once it sectioned, even with plus slides.  

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
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Boulder, CO 80308
(303) 682-3949 office
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liz <@t> premierlab.com
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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of David Burk
Sent: Thursday, March 20, 2014 9:55 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Oil Red O staining question

Experts!

 

Is there any reason you would not consider formalin fixation followed by cryoprotection/cryosectioning for Oil Red O (or bodipy for that matter) staining of mouse muscle? 

I ask as many folks seem to have difficulty in the flash freezing aspect of tissue collection and end up with lots of ice crystal damage. While the above method is still subject to freezing artifact it does at least negate the 'rushed collection' part of the process.

 

The literature favors flash frozen tissue but we've done successful staining of liver that has been fixed then cryoprotected prior to staining and things looked fine. Is there an issue with tissue adhering to slides after sectioning or some other reason this method is not preferred?

 

I value your opinion as I am wondering why one would choose one approach over the other.  Let's pretend tissue antigenicity isn't a factor - only section quality and lipid droplet staining.

 

Thanks for your comments!

 

David

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