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@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 <walkure2...@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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