[Histonet] cutting frozen sections of myocardium

From:Shaumik Adhya

Hi Histonetters,

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.

Shaumik Adhya

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