Preadipocyte Localization


Does anyone have any experience immunohistochemically (or 
histochemically) localizing preadipocytes in muscle tissue?  If so, what 
techniques do you use and what antibodies/stains?
Second, I am having difficulty obtaining sections from frozen muscle 
tissue where the fat cells, muscle fibers, and connective tissue remains 
intact through my immunohistochemical staining procedure. I am currently 
sectioning at 10 micrometers but unfortunately the cellular structure of 
the fat cells is destroyed. I am using poly-L-Lysine coated SuperFrost 
slides. Once sectioned I air dry for 24 hours, fix in PLP, antigen 
retrieve in 0.8 M Urea (10 minutes in the microwave), and subsequently 
perform the immunohistochemical reaction (primary antibody, then use a 
Tyramide Signalling Kit from Molecular Probes for detection, Oil Red 
Stain for lipids, and Hoechst Stain for Nuclei). Does anyone have any 
idea how I can ensure that the section remains fixed to the slide or 
should I expect some loss of tissue?
I appreciate any help I can get. Thank you.
Phillip Huff
FBN Dummerstorf

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