Here, here Gayle, I have a picture on my website of a calcified horse carpal bone I cut using the Instrumedics tape transfer system, I bet no one would have been able to do that without using the tape. www.ihctech.net
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 gayle callis Sent: Monday, November 23, 2009 10:21 AM To: 'Histonet' Cc: emmanuel.mi...@leica-microsystems.com Subject: SPAM-LOW: Reponses to Merced and Richard Re: [Histonet] cryojane tape transfer system You wrote: Unless I'm missing something, I don't understand why people use this tape? It seems like a marketing gimmick to me...ol' fashion' melting of sections onto slides works perfectly for us... ? Regards, Merced Merced, Yes, you are missing something. If you have ever tried to cryosection undecalcified bone or extremely difficult tissues that simply will not result in "ol' fashion' melting" onto a slide , then you would understand why people use this unique cryosectioning system. It is not some "marketing gimmick" but an unique instrument helping many laboratories obtain frozen sections that otherwise are scrunched up, shattered, and destroyed. I suggest you go to the Instrumedics website or www.alphelys.com for a superb slide show and learn how this instrument works before making assumptions about a technology that serves many of us more than well. A happy, informed user of the Cryojane................ Gayle M. Callis HTL/HT/MT(ASCP) Bozeman MT As for what Richard wrote: Hi Everybody, I was hoping to get some advice - I'm cryosectioning plant tissues and transferring sections to slides using the Cryojane system. However, i'm having problems in transferring the sections without them falling apart during the tape transfer. I'm fixing my tissue for 24 hours in ethanol:acetic acid (3:1), embedding in O.C.T, snap-freezing and then sectioning at between 2 and 14 microns. The sections seem to be ok but whenever i remove the adhesive tape from the slide a large part of the tissue is removed with it. As a result I lose the majority of my section. I've tried using both 1x and 1/2x slides (CFSA adhesive slides from instrumedics) but neither have given satisfactory results. Does anyone have any suggestions as to how i could reduce the loss of tissue? Any advice would be much appreciated. Thanks Richard Dear Richard, I could be the fixative you are using that causes the problem. If the fixative contains alcohol, the alcohol acts as antifreeze when you try to snap freeze a tissue, animal or plant. The alcohol may cause problems with how the pink tape sticks to the face of the plant tissue, and allows them to fall apart during the tape transfer. If you rinse away the fixative, then you should cryoprotect the fixed plant tissue with 30% sucrose before snap freezing. vbThis will remove the alcohol. If cryoprotection causes problems with the final staining results, then try unfixed plant tissue, snap freeze, Cryojane tape transfer the section and then fix the transferred plant section in your favorite fixative. You may have to optimize the time in fixative though. Other suggestions: Do a double UV flash, but wait for 15 to 20 seconds between flashes. You must allow the UV light source (capacitor) build up enough charge to work properly. This double flash seems to help polymerize the coating more thoroughly, and the section should transfer better. Also, the tape must be removed at an angle across the slide, very slowly, and inside the cryostat (I am sure you probably do this already.) Also, try the 4X slide if you still have problems with 1/2X and 1X slides. You might ask Leica to send you a few to try before investing in a whole box of these. Contact Emmanuel Mineo, Intrumedics Product Manager emmanuel.mi...@leica-microsystems.com for the 4X slides. Manny is a nice gentleman who has worked with Cryojane for many years and has always been helpful to us. Once again, do the double UV light flash with the 4X slides. They are gooey, but may/should hold more securely. With undecalcified bone, we use the 1/2X but do the double flash routinely for all sections. Good luck Gayle Callis _______________________________________________ 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