I am forwarding this response from a fellow worker, D Kaylor (The techs here have consistently good results when freezing fresh muscles for enzyme histochemistry.
The small holes are freeze artifact. Usually they occur when the muscle is not frozen fast enough. We use methyl butane floating in a liquid nitrogen bath. Either way, the temp is critical. It needs to be at least -155 degrees C. The muscle must be submerged and held under for about 45 seconds to one minute. Do not start or dip the muscle. Once started it must be held submerged. We have never had any issues with humidity. Are the muscle bx swimming in saline? We have better results when they are submitted on saline moistened gauze. We use positively charged slides to pick up our sections. We have some wrinkles but that is due to the sectioning of frozen tissue not the type of slide. Becky Garrison Pathology Supervisor Shands Jacksonville Jacksonville, FL 32209 904-244-6237, phone 904-244-4290, fax 904-393-3194, pager -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Miha Tesar Sent: Saturday, October 15, 2011 4:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Hi! I am working in Institute of Phatology Slovenia EU. Recently I have problems with muscle biopsy. There are a lots of artefact that I can not explain, like little holes in the tissue after the tissue is being frozen. We are using the isopentan and the liquid nitrogen for the freezing. Can enyone give me some ideas haw to avoid this artefact. How important is humidity in laboratory for the good results? The next problem is that after I put the tissue on the microscope slides after coloring the GTK all the tissue gets wrinkeld. Iam using the poly L lisin microscope slides or the Immuno microscope slides. Thx in advance! Best regards Miha from SLO _______________________________________________ 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
