Re: [Histonet] Saturated Fat Staining


If these are already collected and is a retrospective study, it seems like you are pushing the limitations of histochemical analysis.  Unless there are very big physiological differences between control and treated, is hard to see how you can see small differences through a microscope with stains.  You said there was "little variability" with unsaturated fats during quantification.  I assume that is with a stain.  But is there a statistical difference by more precise measurements?  If already done, then no suggestions other than what you've had already and other than good luck.  But in a different life and faced with such kinds of analysis, we just acknowledged histochemical limitations.  Sounds like part of the liver should be cut, weighed, fat extracted, and do something like  gas chromatography analysis.  Then get precise measurements of saturated and un and other lipids.  The human results were surely done with something a bit more precise and quantifiable than an oil red o sta
in on a microscopic section?  Or maybe do the project in parallel, part for quantifiable analysis and part liver for you done somehow in FFPE.  But best wishes for success.

Ray Koelling
PhenoPath Labs
Seattle, WA 

 -------------- Original message ----------------------
From: "Griffin-Reyes, Michelle A" 
> 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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