[Histonet] Frozen section protocol on rat tendon and/or muscle aswell

[Lee & Peggy Wenk]

Is the -80 degrees really a correct temp? We freeze our muscles around -150 
to -160 Degrees C.

Peggy A. Wenk, HTL(ASCP)SLS

-----Original Message----- 
From: Ignacio Ruz Caracuel
Sent: Thursday, January 02, 2014 7:27 AM
To: Peter Petro ; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Frozen section protocol on rat tendon and/or muscle 
aswell

Dear Peter:

Our muscle freeze protocol is very easy. We employed a small piece of cork
of about 1x1cm, we pour a drop of OCT embedding medium and we placed the
muscle on it. It´s important not to pour a big drop of OCT, cause it
creates holes in the muscle as it infiltrates. Then we cooled isopentane in
liquid nitrogen (-80 degrees) for about 10 minutes, we disolved it in case
it aggregates and then we freeze the muscle for about 20 second.

If you don´t pour too much OCT and you achieve a fast freezing with the
isopentane at the correct temperature you won´t have those disturbing
holes.

Best regards,

Ignacio Ruz-Caracuel
Histology Intern Student
Faculty of Medicine, Córdoba, SPAIN
http://www.uco.es/regmus/


2014/1/2 Peter Petro <walkure2009 <@t> gmail.com>

> Dear all,
>
>
> Happy New Year.
>
>
> We are planning to work on rat tendon and frozen section of tendon will be
> performed.  I'd like to ask for a better protocol to preserve, process and
> section tendon as we found there is a lot of ice crystal (holes) on
> sections of muscle that seriously affect our subsequent staining.  Any
> better protocol to freeze muscle is also welcomed. We don't have much
> experience in handling frozen tissues.
>
>
> Best Regards,
>
> Peter
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