Fwd: [Histonet] cutting frozen sections of myocardium

[Atoska S. Gentry]

Hello is it possible for you to fix overnight or for 24hrs and then 
cryoprotect in sucrose/fixative or sucrose/PB before freezing for 
cryosectioning? If you know that fixation won't destroy the enzyme you 
might try this method. Best wishes. Atoska

>Date: Tue, 24 Feb 2004 10:16:44 +0000
>From: Shaumik Adhya <shaumik.adhya <@t> ucl.ac.uk>
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>Subject: [Histonet] cutting frozen sections of myocardium
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>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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Atoska S. Gentry B.S., HT(ASCP)
Research Assistant III
Scott-Ritchey Research Center
College of Veterinary Medicine
Auburn University, AL  36849
Phone# (334)844-5579  Fax# (334)844-5850