We routinely place NBF fixed tissues rinse briefly (often fish livers) in
20% sucrose until it sinks - the sucrose solution removes NBF as it
exchanges - blot and snap freeze. If needed, one could apply a vacuum to
speed up exchange. 

To keep NBF prefixed section on a plus charge slide, air dry 30 minutes
before staining.  The Churukian Oil Red O is superb since it is not gooey,
thick nor difficult to make up and after using this, I never went back to
messy propylene glycol method.   After ORO staining, gently rinse with
water, counterstain using appropriate rinsing steps a light hematoxylin,
blue and mount aqueous.  If you do not have stain procedure, I will be
happy to attach via personal email. 

 

At 08:43 AM 6/12/2003 +1000, you wrote:
>     Sean,   I would rinse the tissue in water (to remove excess formalin),
>impregnate with 30% OCT in water 3-4 hours (or a sucrose substitute), blot
>to remove excess solution and freeze ready for cryotomy.  Use sticky slides
>(Superfrost or equivalent) to mount sections.   The Churukian Oil Red O
>procedure works well.   Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) 
>Laboratory Manager 
>  Westmead, 
>Locked Bag 4001, Westmead, 2145, AUSTRALIA. 
>Tel: (02) 9845 3306 
>Fax: (02) 9845 3318  
>http://www.histosearch.com/homepages/TonyHenwood/default.html 
>http://us.geocities.com/tonyhenwoodau/index.html  
> 
>  -----Original Message----- 
>From: Sean Trombley [mailto:sean.trombley.b@bayer.com] 
>Sent: Thursday, 12 June 2003 5:13 
>To: histonet 
>Subject: Oil Red O  
>  Hi Everyone!     on NBF fixed frozen mouse liver 
>    We are having 
>  Thanks in advance for 
>the help!   Sean 
>Bayer Corp 
>W. Haven, CT 
>(203)812-5471  
>  
> 
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
S. 19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

email: gcallis@montana.edu