Re: [Histonet] Saturated Fat Staining

From:Rene J Buesa



The fats in any FFPE tissue have been partially dissolved by the ante-medium, and Oil Red O will not give very good results. Osmium is too toxic and I would avoid its use.
I think you could do two things:
1- eliminate the ante-medium and use dehydration with 2-propanol, followed by a mixture of mineral oil + 2-propanol as ante-medium (before the paraffin), and
2- try Sudan Black.
 
I also would stain first a group of untreated mice tissues first to get a control of the procedure, and use the treated mice afterward.
Hope this will help you!
René J. 

--- On Fri, 7/11/08, Griffin-Reyes, Michelle A  wrote:

From: Griffin-Reyes, Michelle A 
Subject: [Histonet] Saturated Fat Staining
To: histonet@lists.utsouthwestern.edu
Date: Friday, July 11, 2008, 9:24 AM

I am having some troubles finding a way to be able to stain saturated
lipids in FFPE mouse tissues. Osmium tetroxide and oil red O stain well
but they are only showing unsaturated lipids. My lab is looking to see
if there is a difference in lipid content of treated mice vs. control
mice. So far, we have seen very little variability in the amount of
unsaturated lipids during quantification which leads us to believe there
may be a difference in saturated fats (based off of human results that
we have our mouse model based off of). Does anyone have any suggestions?
Thanks so much for the help!

Michelle A. Griffin-Reyes
Comparative Pathology Laboratory
University of Iowa: College of Medicine



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