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 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet