[Histonet] Oil Red O question

[Kinsley, David]

Hi Andi,

I had always performed oil red O on fresh frozen sections.  I fixed in 10%
NBF after I cut the slides.  I did the oil red o stain, counterstained in
hematoxylin, rinsed in water and let the slides air dry overnight, then
cover slipped with an aqueous mounting medium.  Once the aqueous medium is
dry, the slides can be placed in xylene and permanently mounted.  This way
the lipids which dissolve easily in xylene and to some degree in alcohols
are intact.  If you liver samples are arriving already fixed, you can
cryoprotect overnight in 30% sucrose, then snap freeze in isopentane/dry
ice.  Hope this helps.   

Dave


-----Original Message-----
From: Andrea Grantham [mailto:algranth <@t> u.arizona.edu] 
Sent: Wednesday, November 10, 2004 12:25 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Oil Red O question


This may be a crazy question. I'm doing mouse livers for a lab here and 
they requested Oil Red O. Did the stain - looked great. They are asking if 
it was possible that some of the fat washed away during the washing steps. 
The experiment requires that they quantitate the fat found in these 
sections. Is this a possibility? Is there a better way to quantitate fat?
For the record - my washing steps are done under a gentle stream of tap 
water running into a corner of the stain dish or coplin jar - not a full 
force gush of water from the faucet directly on the slides. Thanks. Andi
Grantham
.....................................................................
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