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.
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