RE: Cryojane Tape Transfer System I have found that you need to have your tape at the same temp as your tissue to have your tissue transfer completely from the tape. I set everything up in the morning putting the tape in the cryostat and get everything equilibrated first for about 2 hours or so. Hope it helps.
-----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of histonet-requ...@lists.utsouthwestern.edu Sent: Monday, November 23, 2009 1:02 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 72, Issue 26 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-requ...@lists.utsouthwestern.edu You can reach the person managing the list at histonet-ow...@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. HT Opening (Marian Powers) 2. cryojane tap transfer system (Richard Pattison) 3. Re: cryojane tap transfer system (Merced M Leiker) 4. Suggestions for Cassette Labeling (Fye Beth) 5. RE: Suggestions for Cassette Labeling (Rathborne, Toni) 6. Reference labs doing parafibromin? (Sebree Linda A) 7. Mouse eyes (Durden, Kelley) 8. Job Opening (Walzer Susan) 9. Reponses to Merced and Richard Re: [Histonet] cryojane tape transfer system (gayle callis) 10. RE: cryojane tap transfer system (CHRISTIE GOWAN) ---------------------------------------------------------------------- Message: 1 Date: Sun, 22 Nov 2009 13:54:28 -0500 From: Marian Powers <mpow...@dpspa.com> Subject: [Histonet] HT Opening To: histonet@lists.utsouthwestern.edu Message-ID: <5d7de0e60911221054i5971a474y64a461eeb5aa8...@mail.gmail.com> Content-Type: text/plain; charset=ISO-8859-1 DPS in Dover, Delaware, is currently seeking a full time histotech, day or evening shift. All inquiries please email mpo...@dpspa.com -- Marian L. Powers, HT(ASCP) Manager, Technical Operations Doctors Pathology Services 1253 College Park Drive Dover, DE 19904 302-677-0000 ------------------------------ Message: 2 Date: Mon, 23 Nov 2009 09:01:39 -0500 From: Richard Pattison <richard.patti...@gmail.com> Subject: [Histonet] cryojane tap transfer system To: Histonet@lists.utsouthwestern.edu Message-ID: <4b0a95c3.4090...@googlemail.com> Content-Type: text/plain; charset=ISO-8859-1; format=flowed 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 ------------------------------ Message: 3 Date: Mon, 23 Nov 2009 09:26:51 -0500 From: Merced M Leiker <lei...@buffalo.edu> Subject: Re: [Histonet] cryojane tap transfer system To: Richard Pattison <richard.patti...@gmail.com>, Histonet@lists.utsouthwestern.edu Message-ID: <5d6d90dc816b8e32d70f0...@cdywxp1931.ad.med.buffalo.edu> Content-Type: text/plain; charset=us-ascii; format=flowed 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 --On Monday, November 23, 2009 9:01 AM -0500 Richard Pattison <richard.patti...@gmail.com> 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 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Merced M Leiker Research Technician III Cardiovascular Medicine 348 Biomedical Research Building State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 USA lei...@buffalo.edu 716-829-6118 (Ph) 716-829-2665 (Fx) No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ------------------------------ Message: 4 Date: Mon, 23 Nov 2009 08:38:05 -0600 From: Fye Beth <beth....@hcahealthcare.com> Subject: [Histonet] Suggestions for Cassette Labeling To: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Message-ID: <938f8ec5a524d34eb5796e23e52781d329a0fca...@nadcwpmsgcms05.hca.corpad.ne t> Content-Type: text/plain; charset="us-ascii" There have been some great suggestions for reducing or catching slide labeling errors. I'm interested in the same for cassette labeling. Currently, we have the patient's last name on one side of the cassette, and the site listed on the other. This makes it easier to identify blocks if they are numbered incorrectly. Beth A. Fye, CT (ASCP) Pathology Technical Manager HCA Richmond Hospital Laboratories office: (804)228-6564 fax: (804)323-8638 <mailto:beth....@hcahealthcare.com> ------------------------------ Message: 5 Date: Mon, 23 Nov 2009 09:41:14 -0500 From: "Rathborne, Toni" <trathbo...@somerset-healthcare.com> Subject: RE: [Histonet] Suggestions for Cassette Labeling To: "Fye Beth" <beth....@hcahealthcare.com>, <histonet@lists.utsouthwestern.edu> Message-ID: <e78340c766a5284d999f5f5891ddf8900baf8...@smcmail.somerset-healthcare.co m> Content-Type: text/plain; charset="utf-8" That seems like a lot of effort. Do your pathologists do the grossing? or PA's? -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu]on Behalf Of Fye Beth Sent: Monday, November 23, 2009 9:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Suggestions for Cassette Labeling There have been some great suggestions for reducing or catching slide labeling errors. I'm interested in the same for cassette labeling. Currently, we have the patient's last name on one side of the cassette, and the site listed on the other. This makes it easier to identify blocks if they are numbered incorrectly. Beth A. Fye, CT (ASCP) Pathology Technical Manager HCA Richmond Hospital Laboratories office: (804)228-6564 fax: (804)323-8638 <mailto:beth....@hcahealthcare.com> _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE This message and any included attachments are from Somerset Medical Center and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or trade secret information entitled to protection and/or exemption from disclosure under applicable law. Unauthorized forwarding, printing, copying, distribution, or use of such information is strictly prohibited and may be unlawful. If you are not the addressee, please promptly delete this message and notify the sender of the delivery error by e-mail or you may call Somerset Medical Center's computer Help Desk at 908-685-2200, ext. 4050. -------------------------------------------------------------- Somerset Medical Center is the recipient of the 2009 Orthopedic Surgery Excellence Award(tm) from HealthGrades, the nation's leading health care ratings company. Visit Somerset Medical Center's Web site - www.somersetmedicalcenter.com - for news, event listings, health information and more. Join the Discussion: Facebook: www.somersetmedicalcenter.com/fb Twitter: www.twitter.com/SomersetMedCtr ------------------------------ Message: 6 Date: Mon, 23 Nov 2009 09:35:07 -0600 From: "Sebree Linda A" <lseb...@uwhealth.org> Subject: [Histonet] Reference labs doing parafibromin? To: "Histonet" <histo...@pathology.swmed.edu> Message-ID: <8c023b4ab999614ba4791baeb26e27381bf...@uwhc-mail01.uwhis.hosp.wisc.edu> Content-Type: text/plain; charset="us-ascii" Good Monday morning, One of our pathologists is getting pressure from the endocrinologists to bring on Parafibromin (gene: hrpt2). Are there any reference labs out there offering this? Thanks, Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 ------------------------------ Message: 7 Date: Mon, 23 Nov 2009 10:44:21 -0500 From: "Durden, Kelley" <kelleydur...@pathology.ufl.edu> Subject: [Histonet] Mouse eyes To: "'histonet@lists.utsouthwestern.edu'" <histonet@lists.utsouthwestern.edu> Message-ID: <92e6b93e0a3d544c87ddde33e7608aae0956c...@hsc-cms01.ad.ufl.edu> Content-Type: text/plain; charset="us-ascii" My question concerns mouse eyes. Can anyone send me their fixation suggestions? We are receiving mouse eyes that end up with a "wavy" look instead of a nicely defined "C" and have no explanation for this occurrence. Can anyone send me their mouse eye processing schedule? We have a protocol that has proved fast and true but would welcome other suggestions. Can you give me a good idea for making sure sections stick to slides well? Gold Plus? What else? Has anyone experienced a retinal detachment after or upon staining? Retina attached after sectioning - detached after staining - just routine H&E. Should I heat for an extended period of time? Kelley Durden, HT ASCP University of Florida Molecular Pathology Core ------------------------------ Message: 8 Date: Mon, 23 Nov 2009 09:48:19 -0600 From: Walzer Susan <susan.wal...@hcahealthcare.com> Subject: [Histonet] Job Opening To: "histo...@pathology.swmed.edu" <histo...@pathology.swmed.edu> Message-ID: <4bf03f5404ebde409af9232da74b9ded2aeab5e...@fwdcwpmsgcms09.hca.corpad.ne t> Content-Type: text/plain; charset="us-ascii" We have an opening for a tech, days no weekends at St Pete.General Hospital in St Pete,FL. Call our HR dept at 727 384-1414 X 4814. ------------------------------ Message: 9 Date: Mon, 23 Nov 2009 10:20:58 -0700 From: "gayle callis" <gayle.cal...@bresnan.net> Subject: Reponses to Merced and Richard Re: [Histonet] cryojane tape transfer system To: "'Histonet'" <histonet@lists.utsouthwestern.edu> Cc: emmanuel.mi...@leica-microsystems.com Message-ID: <000101ca6c61$541e4d60$fc5ae8...@callis@bresnan.net> Content-Type: text/plain; charset="us-ascii" 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 ------------------------------ Message: 10 Date: Mon, 23 Nov 2009 17:22:40 +0000 From: CHRISTIE GOWAN <christiego...@msn.com> Subject: RE: [Histonet] cryojane tap transfer system To: <lei...@buffalo.edu>, <richard.patti...@gmail.com>, <histonet@lists.utsouthwestern.edu> Message-ID: <snt122-w392b7d2052613e9c485796ae...@phx.gbl> Content-Type: text/plain; charset="iso-8859-1" Hi Richard, I am assuming that you are using the flash in the cryojane system to adhere the specimen to the slide. Make sure your tape is not too old and don't store it in the cryostat. It sounds like you are able to get the section but unable to get it to adhere. Make sure you pass your roller over the tape and slide several times before putting it under the flash and always use a positive charged slide from a freshly opened box. Good luck. I hope this helps a little Christie ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 72, Issue 26 **************************************** Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates Direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. 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