[Histonet] cutting frozen sections of myocardium
I'm a clinician trying to get to grips with histotechnology techniques for
research. I'm having a lot of difficulty cutting frozen sections of rat
myocardium. I seem to be cutting sections with a lot of holes in them, and
the fibres seem poorly arrayed. I'm cutting rat myocardium to practice before
human myocardial biopsies.
The rats are killed, the hearts harvested, without fixation. They're cut into
small pieces before staining for 20 hours in senescence associated beta
galactosidase solution (essentially x-gal solution at pH 6). They are then
moutned into OCT, frozen in isopentane and cut on a cryotome at -20
centigrade. The slides are then fixed in 3% paraformaldehyde, wawshed in
distilled water, before Haemotxylin & Eosin staining.
Any advice, tips or tricks will be much appreciated.
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