Especially for bone, I have found that formalin fixation and then infiltrating in 30% sucrose before freezing the way Samuri described works the very best for me.
Regards, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. Ste.215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 www.ihctech.net www.ihcrg.org -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Robert Richmond Sent: Thursday, November 19, 2009 11:22 AM To: histonet@lists.utsouthwestern.edu Subject: SPAM-LOW: [Histonet] Re: freezing skeletal muscle Galina Deyneko asks about freezing skeletal muscle. I'm not sure that temperature control of the isopentane is too important - it should be very slightly viscous. But do get rid of explosive isopentane and get a non-flammable substitute - see my previous posts on this topic. The technique for freezing skeletal muscle is easier demonstrated than described. Hold a small piece (maybe 5 x 10 mm) in tweezers, and coat it with talc powder so that it appears white on the surface. Then dip the specimen in and out of the isopentane - roughly two dips per second - until it is frozen solid. That will eliminate the ice crystal artifact. But you have to try it a few times before it will quite work for you. Set aside one dead rat and one afternoon. Bob Richmond Samurai Pathologist Knoxville TN _______________________________________________ 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
