[Histonet] Unsuccessful Muscle Lipid Staining

From:Sonny Duong

Hi all,

I have been trying to stain mouse muscle tissue for 
intramuscular triglyceride droplets with Oil Red O 
(dissolved in triethyl phosphate) for some time now, and my 
results have been disastrous.  I've tried it on both fresh 
frozen and formaldehyde fixed tissues (and cryoprotected) 
cryosections, but almost always it seems that the lipid 
droplets within the fibers (maybe?) leak out of the cells 
and conglomerate on the edges of the tissue and within gaps 
between the cells.  Very few of the fibers, if any, display 
any lipid droplet staining and it is always very weak. The 
lipid droplets all conglomerate in roughly the same 
positions between serial sections as well, which leads me 
to believe something is happening during the 
freezing/fixation process, or the cutting in the 
cryosection.  Does anyone have any suggestion as to what I 
could be doing wrong here (i.e cutting technique, 
temperature, fixatives)?  Also, does anyone have any 
experience with using ORO in triethyl phosphate?

Thank you all for your time,
Sonny Duong
University of Virginia
Green Lab

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