I use isopentane suspended in liquid nitrogen, too. sorry if this sounds curt, but due to your cut-and-pasting, im not entirely sure what point youre trying to get at...?
________________________________ From: Akemi Allison-Tacha <akemiat3...@yahoo.com> To: Jennifer MacDonald <jmacdon...@mtsac.edu>; Ingles Claire <cing...@uwhealth.org>; Bernie Taupin <bernietau...@ymail.com> Cc: Histonet@lists.utsouthwestern.edu; histonet-boun...@lists.utsouthwestern.edu Sent: Wednesday, April 8, 2009 1:03:04 AM Subject: Re: [Histonet] "FREEZY" spray Bernie, I used to use the method below for FS on muscles at Emanual Hospital in Portland OR. Since we had such wonderful success, we incorporated this method for all FS. Years later, I developed the "MATSSE (pH3.4) 1-Step Trichrome Stain Kit" (Modified Trichrome Method for Muscle Biopsies Cut on Frozen Sections) for Biocare Medical. This kit has long since discontinued by the way....This came from Biocare's Data Sheet. NOTE:Muscle tissue should be suitably obtained and frozen within 30 minutes after excision. The usual method of freezing is to first mount a transverse section of the muscle on a chuck using 10% tragacanth gum or equivalent commercial frozen section mounting media as the adhesive. The chuck with the mounted specimen is then immersed in prechilled isopentane until frozen. Isopentane is precooled when a container of the substance is placed into liquid nitrogen. At a temperature of -160° C, the isopentane has a slightly syrupy consistency. Care should be taken to remove the muscle sample when freezing is complete, usually after 25 to 30 seconds. Too short a freezing time produces artifacts; prolonged freezing can produce cracking of the block. NOTE: If the sample cannot be sectioned immediately after freezing, it may be wrapped in foil and placed in a plastic-lidded container along with a small amount of ice for moisture. Specimens may be stored at -70° C until sectioned. Regards, Akemi Akemi Allison-Tacha BS, HT (ASCP) HTL Histology Manager Associated Pathology Medical Group Laboratories 105A Cooper Court, Los Gatos, CA 95032 Cell: (425) 941-4287 E-Mail: akemiat3...@yahoo.com --- On Tue, 4/7/09, Bernie Taupin <bernietau...@ymail.com> wrote: From: Bernie Taupin <bernietau...@ymail.com> Subject: Re: [Histonet] "FREEZY" spray To: "Jennifer MacDonald" <jmacdon...@mtsac.edu>, "Ingles Claire" <cing...@uwhealth.org> Cc: Histonet@lists.utsouthwestern.edu, histonet-boun...@lists.utsouthwestern.edu Date: Tuesday, April 7, 2009, 9:06 PM > Does this apply to Liquid Nitrogen as well? I'd hate to have to try to cut > fatty sections in the cryostat without it! I do mostly cryo, and I've never used liquid nitrogen to chill anything within the cryostat. What's that all about? I find that it leaves to many ice crystal artifacts anyway. Not cold enough fast enough. You might be making it harder on yourself... just set the temperature of the cryostat for the appropriate temperature of whatever tissue youre cutting, and you should have no need for a crutch, whether it be fluoroethane or liquid nitrogen. kisses, flowers and rainbows, Bernie Taupin, King of Cryomicrotomy, Esq. _______________________________________________ Histonet mailing list histo...@lists.utsouthwestern..edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet