[Histonet] Oil red O stain on formalin fixed liver

Tony Henwood (SCHN) tony.henwood <@t> health.nsw.gov.au
Mon Jan 28 16:21:08 CST 2013


(From Doug Reye's old Laboratory).

I have found the following technique the best:

Cryotomy of Formalin Fixed Tissues
Sometimes frozen sections are requested on tissue that has been fixed in 10% formalin. Cutting frozen sections of formalin fixed tissue can be frustrating at times due to shredding of the tissue or failure of the sections to stay on the slides. The following procedure makes sectioning a lot easier:


1.	OCT frozen embedding compound (or equivalent)
2.	OCT Infiltration Solution:
	OCT Compound	4ml
	Distilled Water		8ml     
3.	Sticky Slides (Poly Lysine or equivalent)


1.	Take well-fixed tissue and gently wash in tap water   1hr.
2.	Place in OCT Infiltration Solution and place on rotor.
	*	For small pieces (needle or thin wedge) 2hr.
	*	For larger pieces 4hrs - overnight.
3.	Gently blot excess solution from tissue and embed in OCT and freeze.
4.	Cut frozen sections and pick up on adhesive coated slides.
5.	Place one slide in Methanol and stain H&E or air-dry sections for oil red O staining

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager & Senior Scientist 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Dmborel
Sent: Tuesday, 29 January 2013 6:10 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Oil red O stain on formalin fixed liver

We have an autopsy on a young child.  I have a working diagnosis of possible Reye's syndrome and need an Oil Red O stain on a section of formalin fixed liver (not processed) to confirm microvesicular hepatic steatosis.  Does anyone do this stain, and could I send the tissue or unstained slides for you to perform the stain and send islides back for me to interpret?  I can provide a bone marrow smear to use as a positive control slide, pay shipping both directions and a technical fee.

Thank you,

David M. Borel, M.D.
Pathology Services, P.A.
Topeka, KS
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