[Histonet] Diff Quik II
Good Morning! I was wondering who out there is using the Diff Quik II stain for H. pylori? I am having a hard time finding where to buy it! Do you just buy the whole DQ kit or do you just buy Diff Quik II solution? Any suggestions would be great! Thanks! Happy Friday! Jessica Piche-Grocki, HT(ASCP) Waterbury Hospital ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] reticulin stain
Derek: I am sending it to you under separate cover. René J. --- On Fri, 6/19/09, Derek Papalegis derek.papale...@tufts.edu wrote: From: Derek Papalegis derek.papale...@tufts.edu Subject: [Histonet] reticulin stain To: histonet@lists.utsouthwestern.edu Date: Friday, June 19, 2009, 9:11 AM I am having some difficulty getting the proper results from my reticulin stain. I have been following and tweaking the procedure outlined in Carson's book without any success. Could someone please forward me their procedure that has all the kinks worked out? Thanks, Derek -- Derek Papalegis HT (ASCP) Senior Histology Technologist Division of Laboratory Animal Medicine Tufts University 136 Harrison Avenue Boston, MA 02111 phone: 617 636-2971 fax: 617 636-8354 ___ 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
RE: [Histonet] Diff Quik II
I purchase Diff Quik Solution II in a 2.5L bottle from Cardinal Health Catalog # B4132-12A. It's not something that they have in stock and it usually ships from Dade Behring Inc. which is who makes it. It usually takes me a couple of weeks to get it, but I have no problem receiving it. Jodie Robertson, HT (ASCP) QIHC Pathology Sciences Medical Group -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jessica Piche Sent: Friday, June 19, 2009 5:52 AM To: histonet Subject: [Histonet] Diff Quik II Good Morning! I was wondering who out there is using the Diff Quik II stain for H. pylori? I am having a hard time finding where to buy it! Do you just buy the whole DQ kit or do you just buy Diff Quik II solution? Any suggestions would be great! Thanks! Happy Friday! Jessica Piche-Grocki, HT(ASCP) Waterbury Hospital ___ 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
[Histonet] Re: anti Brdu
We have used mouse anti-Brdu from Becton Dickinson for years. It is still available from BD BioSciences Cat. 347580. I know it is made in mouse but we used an IgG1 secondary and always had good results. There may be some cytoplasmic label, probably background, but you are interested in the nuclear staining and it is very distinct. We have used it on both frozen and paraffin processed tissues. Donna Reynolds, HT (ASCP) Dept. Cancer Biology U.T. M.D. Anderson Cancer Center, Houston Tx Message: 3 Date: Wed, 17 Jun 2009 14:26:09 -0400 From: Madary, Joseph mada...@medimmune.com Subject: [Histonet] source for rat anti mouse brdu? To: histonet@lists.utsouthwestern.edu Message-ID: e5b1c3d1305efd4c9ca7f63094135f5e0226a...@md1ev002.medimmune.com Content-Type: text/plain; charset=us-ascii I have not done a brdu on mouse in five years. Used Harlan but they are not selling that anymore. I used a procedure using rat anti mouse brdu primary, rabbiot anti rat mouse adsorbed secondary, then ABC/DAB. Anyone have any idea on a good source? I used biocompare and found stuff but would like to hear comments. Also anyone use conjugated brdu? Nick Madary, HT/HTL(ASCP)QIHC Medimmune Histology Mgr, OMW, Area 4, Lab 2438 301.398.4745(vm) 301.398.6360(lab) 301.398.9745(fax) * ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Ventana Reagent Cost??
Hello everyone, I am looking for a cost of some of the Ventana reagents can anyone help? Specifically: LCS: CC1: CC2: EZ Prep: Reaction Buffer: I am not currently a customer so I didn't want to phone Ventana directly if I don't have to. Thanks Tiana This email and/or any documents in this transmission is intended for the addressee(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original. Ce courriel et tout document dans cette transmission est destiné à la personne ou aux personnes à qui il est adressé. Il peut contenir des informations privilégiées ou confidentielles. Toute utilisation, divulgation, distribution, copie, ou diffusion non autorisée est strictement défendue. Si vous n'êtes pas le destinataire de ce message, veuillez en informer l'expéditeur immédiatement et lui remettre l'original. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Ventana Reagent Cost??
`If you don't use their auto stainer probably you will not be able to use their antibodies that, as far as I know, are optimized to be used in their instrument in what they call liquid cover slips for room and high temperature. René J. --- On Fri, 6/19/09, Tiana Fountain tfount...@exchange.hsc.mb.ca wrote: From: Tiana Fountain tfount...@exchange.hsc.mb.ca Subject: [Histonet] Ventana Reagent Cost?? To: histonet@lists.utsouthwestern.edu Date: Friday, June 19, 2009, 11:42 AM Hello everyone, I am looking for a cost of some of the Ventana reagents can anyone help? Specifically: LCS: CC1: CC2: EZ Prep: Reaction Buffer: I am not currently a customer so I didn't want to phone Ventana directly if I don't have to. Thanks Tiana -Inline Attachment Follows- This email and/or any documents in this transmission is intended for the addressee(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original. Ce courriel et tout document dans cette transmission est destiné à la personne ou aux personnes à qui il est adressé. Il peut contenir des informations privilégiées ou confidentielles. Toute utilisation, divulgation, distribution, copie, ou diffusion non autorisée est strictement défendue. Si vous n'êtes pas le destinataire de ce message, veuillez en informer l'expéditeur immédiatement et lui remettre l'original. -Inline Attachment Follows- ___ 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
Re: [Histonet] Ventana Reagent Cost??
Ventana prices will vary dependednt upon your (not you per se, but rather your instrument) consumption. Here is what we pay: LCS $74 CC1 $832 CC2 $895 EZprep (10X) $452 Reaction Buffer$65 If you need more info, please contact me Thanks Dusko Trajkovic Pfizer Inc La Jolla From: Burton, Lynn lynn.bur...@illinois.gov To: Tiana Fountain tfount...@exchange.hsc.mb.ca; histonet@lists.utsouthwestern.edu Sent: Friday, June 19, 2009 9:46:37 AM Subject: RE: [Histonet] Ventana Reagent Cost?? The prices I have are on the list you sent. I couldn't find the CC2 price quickly. Lynn Burton Lab Assoc. I Animal Disease Lab Galesburg, Il 61401 From: histonet-boun...@lists.utsouthwestern.edu on behalf of Tiana Fountain Sent: Fri 6/19/2009 10:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Ventana Reagent Cost?? Hello everyone, I am looking for a cost of some of the Ventana reagents can anyone help? Specifically: LCS:$98.40 CC1:$565.80 CC2: EZ Prep:$37.80 Reaction Buffer:$43.80 I am not currently a customer so I didn't want to phone Ventana directly if I don't have to. Thanks Tiana ___ 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
[Histonet] Microtome - Free!
I have a pretty old microtome to donate. It is a Leitz 1512. I also have some of those big blades that it uses. All you have to do is pay for the shipping. Cheers, Maggie Margaret E. Bisher Electron Microscopy Histology Core Facility Manager Department of Molecular Biology Princeton University Moffett Laboratory, Room 113 Princeton, New Jersey Office: (609) 258-7026 Fax: (609) 258-8468 mbis...@princeton.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: retic stain
I too will send you ours to your e-mail. Matt Lunetta HT (ASCP) Message: 13 Date: Fri, 19 Jun 2009 06:26:34 -0700 (PDT) From: Rene J Buesa rjbu...@yahoo.com Subject: [Histonet] reticulin stain To: histonet@lists.utsouthwestern.edu, Derek Papalegis derek.papale...@tufts.edu Message-ID: 431812.3131...@web65714.mail.ac4.yahoo.com Content-Type: text/plain; charset=iso-8859-1 Derek: I am sending it to you under separate cover. René J. --- On Fri, 6/19/09, Derek Papalegis derek.papale...@tufts.edu wrote: From: Derek Papalegis derek.papale...@tufts.edu Subject: [Histonet] reticulin stain To: histonet@lists.utsouthwestern.edu Date: Friday, June 19, 2009, 9:11 AM I am having some difficulty getting the proper results from my reticulin stain. I have been following and tweaking the procedure outlined in Carson's book without any success. Could someone please forward me their procedure that has all the kinks worked out? Thanks, Derek -- Derek Papalegis HT (ASCP) Senior Histology Technologist Division of Laboratory Animal Medicine Tufts University 136 Harrison Avenue Boston, MA 02111 phone: 617 636-2971 fax: 617 636-8354 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] LR White and immunofluorescence
Happy Friday Everybody, I have a user who has a precious sample and wants to embed in LR White, maybe. I'm having him practice on something not precious first. My questions are: 1. He wants to do immunofluorescence and morphology on these nerve samples. He is planning on perfusing in 2% PFA, 0.1% glut in 0.1M sodium cacodylate. Then he will dissect out the nerve, cut it in half and place half in 2% PFA, 0.1% glut (embed this in LR White) and the other half in 4% PFA, 2.5% glut (embed this in Epon) to finish the fixation. Will that work? Or will the perfusion fixation with the weaker fix cause the stronger submersion fix to be moot? 2. He wants to do immunofluorescence on the LR white sample. I figure, primary antibody, then secondary with glow is no different that secondary with gold ball, am I correct in this? Thanks and have a great weekend. All aglow waiting for your answer, Paula :-) Paula Sicurello VA Medical Center San Diego Veterans Medical Research Foundation (VMRF) Core Research Imaging Center (CRIC) 3350 La Jolla Village Dr., MC151 San Diego, CA 92161 858-552-8585 x2397 Your images flow through our CRIC. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Diff-Quik II
To stain Helicobacter, you need only the Diff-Quik II solution. This is a brand name. I've used several generic equivalents and they work also. Bob Richmond Samurai Pathologist Knoxville TN ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Electron Microscopy
Hi, I am trying to get a a pre-embedding immuno to work. I have tried several different things (fixatives, incubation times, +/- detergent) and it seems that definitely I need to have a detergent present. I used 0.05% Tween20, but the quality of the EM is really bad (it is impossible to distinguish anything from them). Does anybody have experience doing this? Any protocol I could try? Suggestions? Thanks very much in advance for any help. Diego. -- Diego J. Rodriguez Gil Postdoctoral Associate Department of Neurosurgery Yale University School of Medicine PO Box 208082 333 Cedar St, FMB-430 New Haven CT, 06520-8082 U.S.A. Phone: 203-785-4230 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Section Mounting Techniques
Hi everyone, Does anyone have a good technique for mounting bone tissue (rabbit calvaria embedded in plastic) to coated slides? I am having problems with wrinkles following the rolling of the sections onto the slides. I would greatly appreciate any help you might be willing to offer. Thanks. Jim ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Peloris Tissue Processor
Does anyone use Formula 83 as their Clearant on a Peloris tissue processor?? If so, have you had any problems?? Please let me know as we are experiencing consistent problems with water in the Formula 83. Thank you, Ann ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Diff Quik II
We buy Diff Quik II by the gallon directly from Dade. You don't need the kit. Judy Keller, HTL South County Hospital Wakefield, RI From: histonet-boun...@lists.utsouthwestern.edu on behalf of histonet-requ...@lists.utsouthwestern.edu Sent: Fri 6/19/2009 1:01 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 67, Issue 19 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. Synaptophysin (Benny Babinsack) 2. biocare bcl 6 (Orr, Rebecca) 3. Cytokeratin 18 M30 apoptosis staining (Popp, Laurie A.) 4. RNA In Situ Hybridizaton (Bell, Pat) 5. FW: Cytokeratin 18 M30 apoptosis staining (Popp, Laurie A.) 6. Glypican-3 on BondMax (Steven Wilkes) 7. Glypican-3 on BondMax (Steven Wilkes) 8. Glypican-3 on BondMax (Steven Wilkes) 9. xylene and frozen sections (Lynn Dike) 10. Temp work (Loralei Dewe) 11. Diff Quik II (Jessica Piche) 12. reticulin stain (Derek Papalegis) 13. reticulin stain (Rene J Buesa) 14. RE: Diff Quik II (Jodie Robertson) 15. Re: anti Brdu (Reynolds,Donna M) 16. Ventana Reagent Cost?? (Tiana Fountain) 17. Good Used Equipment for Sale (Denise Van Eaton) 18. Mouse necropsy question (Thach, Dzung (NIH/NIAID) [E]) 19. Re: Ventana Reagent Cost?? (Rene J Buesa) 20. RE: Ventana Reagent Cost?? (Burton, Lynn) -- Message: 1 Date: Mon, 15 Jun 2009 11:54:04 -0400 From: Benny Babinsack bbabins...@genesishcs.org Subject: [Histonet] Synaptophysin To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Message-ID: of8f32beea.eb47166f-on852575d6.00575917-852575d6.00575...@genesishcs.org Content-Type: text/plain; charset=ISO-8859-1 We use a IVD from Cellmarque and use pa ncreas for the control Benny Ba Lab Services Senior Tech,n (740)454-5587 ***Confidentiality Notice*** The information contained in this e-mail message ([Histonet] Synaptophysin), including any attachments, are intended only for use of the individual or entity named above (histonet@lists.utsouthwestern.edu). This e-mail may contain information that is privileged, confidential and/or otherwise exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, any disclosure, dissemination, distribution, copying or other use of the communication or its substance is prohibited. If you have received this e-mail in error, please reply to this e-mail indicating you are not the intended recipient and immediately destroy all copies of this e-mail. Receipt by anyone other than the intended recipient is not a waiver of any privileged information. -- Message: 2 Date: Tue, 16 Jun 2009 12:29:47 -0500 From: Orr, Rebecca r...@northshore.org Subject: [Histonet] biocare bcl 6 To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu Cc: 'jpet...@bostwicklaboratories.com' jpet...@bostwicklaboratories.com Message-ID: b44ca3426a2c084f9ba90b93d528159be4e2c...@exch34.enhnet.org Content-Type: text/plain; charset=us-ascii Message: 3 Date: Mon, 15 Jun 2009 15:34:33 -0400 From: Justin Peters jpet...@bostwicklaboratories.com Subject: [Histonet] BCL-6 To: histonet@lists.utsouthwestern.edu Message-ID: 24d22de9e488aa43bf92a4389f2ddb1f05dfa...@mail1.bostwick.com Content-Type: text/plain; charset=us-ascii I am having trouble getting bcl-6 antibody to work (Biocare CM223). Does anyone have a good protocol for me to try? Hi Justin, I run BCl-6 from concentrate 1:50 with Biocare's RED diluent with a Decloaker using RED diluent. Most antibodies perform well with the green diluent; this particular antibody is best using the RED. I also use Mach4 detection... if problems persist, contact me offline, I may be able to help with some other suggestions. Hope this helps. Becky Orr CLA,HT(ASCP)QIHC Technical Specialist Anatomic Pathology NorthShore University HealthSystem 847-570-2771 * -- Message: 3 Date: Tue, 16 Jun 2009 15:59:46 -0500 From: Popp, Laurie A. popp.lau...@mayo.edu Subject: [Histonet] Cytokeratin 18 M30 apoptosis staining To: histonet@lists.utsouthwestern.edu Message-ID: 1488342adb74ec4e969c48bdcde0bd98fb7...@msgebe23.mfad.mfroot.org Content-Type: text/plain; charset=US-ASCII Has anyone worked with this particular antibody? Can you give me any
[Histonet] Superfrost plus gold slides vs superfrost plus
Hi everyone, I'm curious if anyone has tried using Superfrost plus gold slides, are they any better than the regular superfrost plus? I'm dealing with frozen lymph node on a Ventana discovery, I've tried a few different fixing/drying combinations but I still have tissue lifting (not completely but enough to be bothersome!). Wondering if it's worth giving the gold slides a go. Thanks, Katy ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Superfrost plus gold slides vs superfrost plus
I had a group here who were having trouble with their cryo-sections sticking to the slides. Their samples were rat brains. He found that the Gold Super Frost Plus worked well with in-situ but not for immuno. At least that is the comment he wrote down and put on our bulletin board in the lab. Most of the people in the lab tend to use the Super Frost Plus (not the Gold) and they seem pretty happy with them. Margaret E. Bisher Electron Microscopy Histology Core Facility Manager Department of Molecular Biology Princeton University Moffett Laboratory, Room 113 Princeton, New Jersey Office: (609) 258-7026 Fax: (609) 258-8468 mbis...@princeton.edu On 6/19/09 4:26 PM, Milne, Katy kmi...@bccancer.bc.ca wrote: Hi everyone, I'm curious if anyone has tried using Superfrost plus gold slides, are they any better than the regular superfrost plus? I'm dealing with frozen lymph node on a Ventana discovery, I've tried a few different fixing/drying combinations but I still have tissue lifting (not completely but enough to be bothersome!). Wondering if it's worth giving the gold slides a go. Thanks, Katy ___ 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
RE: [Histonet] Superfrost plus gold slides vs superfrost plus
But doesn't the gelatin's protein interfere with immuno results? That's why we do not use glue, gelatin, or any adhesive on immuno cases. Maria Katleba HT (ASCP), MS, MBA Pathology Dept. Mgr Queen of the Valley Medical Center Pathology Laboratory Napa Ca -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Susan Bachus Sent: Friday, June 19, 2009 1:42 PM To: Peggy Bisher; Milne, Katy; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Superfrost plus gold slides vs superfrost plus Good old fashioned gelatin subbing is very cheap works great--I have never seen tissue separate from a subbed slide! Susan - Original Message - From: Peggy Bisher mbis...@princeton.edu To: Milne, Katy kmi...@bccancer.bc.ca; histonet@lists.utsouthwestern.edu Sent: Friday, June 19, 2009 3:34 PM Subject: Re: [Histonet] Superfrost plus gold slides vs superfrost plus I had a group here who were having trouble with their cryo-sections sticking to the slides. Their samples were rat brains. He found that the Gold Super Frost Plus worked well with in-situ but not for immuno. At least that is the comment he wrote down and put on our bulletin board in the lab. Most of the people in the lab tend to use the Super Frost Plus (not the Gold) and they seem pretty happy with them. Margaret E. Bisher Electron Microscopy Histology Core Facility Manager Department of Molecular Biology Princeton University Moffett Laboratory, Room 113 Princeton, New Jersey Office: (609) 258-7026 Fax: (609) 258-8468 mbis...@princeton.edu On 6/19/09 4:26 PM, Milne, Katy kmi...@bccancer.bc.ca wrote: Hi everyone, I'm curious if anyone has tried using Superfrost plus gold slides, are they any better than the regular superfrost plus? I'm dealing with frozen lymph node on a Ventana discovery, I've tried a few different fixing/drying combinations but I still have tissue lifting (not completely but enough to be bothersome!). Wondering if it's worth giving the gold slides a go. Thanks, Katy ___ 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 No virus found in this incoming message. Checked by AVG - www.avg.com Version: 8.5.339 / Virus Database: 270.12.80/2187 - Release Date: 06/19/09 06:53:00 Notice from St.Joseph Health System: Please note that the information contained in this message may be privileged and confidential and protected from disclosure. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Superfrost Gold versus Superfrost Plus Charge
You did not say whether the tissues were prefixed with formalin or fresh tissues, frozen then cryosectioned? If prefixed, cryoprotected, frozen and cryosectioned, the Plus Gold should work. They are certainly more expensive! A colleague uses Plus Gold for prefixed, cryoprotected frozen sections destined for immunostaining. She sections, then air dries for a time. You may need to determine the time of drying, even if the tissue is fresh. There are package inserts that tell you HOW to use these slides, both Gold or Plus Charge, at least the ones manufactured by Erie have an insert. The Gold slides should be dried flat, but that may be due to contact with a room temperature surface e.g. tray. To help you air dry, purchase a cheap, small fan and dry any FS on Gold or Plus Charge - make sure the air blows pretty much directly onto the sections whether you lay them flat or slanted to be in airflow. You will hasten the drying time and dry more efficiently even in a 30 minute time frame. Try and get a Gold Plus samples from Erie/Thermo Scientific rep in your area, hopefully they still do this. Superfrost Plus Charge (Erie trademarked) works very well with fresh tissue frozen sections, followed by fixation. If we section fresh tissue, air dry a minimum of 30 minutes, solvent fix and stain, we do not lose sections but we also do NOT use an automated staining system. However, if you fix with NBF and use a retrieval method (MW or enzyme digestion) the prefixed or NBF fixed fresh frozen sections may lift off. We never use any kind of gelatin or glue as an adhesive for immunostaining work, just the Plus Charge (trademarked, meaning Erie manufacturered). I have heard the Mercedes coated slides work very well with prefixed tissue frozen sections. You may want to try those too. Since frozen sections are notorious for being more delicate, they cannot withstand some mechanical manipulations such as the reagent dispensing on a stainer - maybe too forceful??? Maybe that can be adjusted so it still allows good flow but not quite so harsh?? If you cut fresh frozen sections, air dry, fix with 4C acetone for 10 min, dry to evaporate solvent and stain, you can stain. A common complaint is poor morphology, but you can post fix a totally IHC frozen section in NBF for 10 min, rinse, then counterstain with hematoxylin. Dr. Chris van der Loos does this with great success and improves the morphology on his section by doing so. Good luck Gayle M. Callis HTL(ASCP)HT,MT Bozeman MT 59715 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Superfrost plus slides
You did not say which slides you use, but you may want to try Thermo Scientific EXCELL slides. The special coating resists the effects of high pH EDTA antigen retrieval and heat. Hopefully, you are putting any adhesives in the water-bath when using Plus Charge or any other coated slides. The gelatin in these adhesives coats the plus charge and negates that charge. Gayle M. Callis HTL(ASCP)HT,MT Bozeman MT -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Haviland,Joie Sent: Friday, June 19, 2009 5:06 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Superfrost plus slides Hello Histonetters: I am having problems with tissue just disappearing in my lastest batches of slides. I have been doing a lot of HE's and did not have too many problems. But I am cutting testis to use with a CD168 ab. Everytime after the 1mM EDTA AR,the tissue disappears from the slide. I allow the sections to dry overnight in our hood before placing the slides at 60'C overnight. And sometimes I am in a hurry to get it done that I put them right away in the oven. I am also having problems with mouse skin tissue that has hair on it. Seems that the section is lifting on some parts of the slide but not others. It is very frustrating right now. Has anyone else edxperienced this? I have been using the same batch of slides for a couple of weeks Thanks Joie ___ 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
RE: [Histonet] Acid Alcohol
We have used 0.5% to 1% HCl in 70% alcohoL but only for Harris regressive hematoxylin staining. For progressive hematoxylins, and Gills, 1% acetic acid was preferred. Gayle M. Callis HTL(ASCP)HT,MT -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Joseph Saby Sent: Friday, June 19, 2009 5:59 PM To: thisis...@aol.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Acid Alcohol 1% Nitric acid in 70% ethanbol. Joe Saby From: thisis...@aol.com thisis...@aol.com To: histonet@lists.utsouthwestern.edu Sent: Friday, June 19, 2009 5:01:48 PM Subject: [Histonet] Acid Alcohol I have lost the recipe for Acid Alcohol (used to decolorize HE's).? Can someone send it to me? Thanks, Ann ___ 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
FW: [Histonet] xylene and frozen sections
Lynn From: lynn...@hotmail.com To: lynn...@hotmail.com Subject: RE: [Histonet] xylene and frozen sections Date: Thu, 18 Jun 2009 19:53:52 -0500 Is there any literature that says xylene will fade progressive staining HE (using Surgipath Harris Hematoxylin) over a period of years? Our frozen staining series after the eosin has two 95% ROH's and six 100% and then clear in Propar. I mistakenly replaced the last three 100% ROH with xylene when I rotated the series. I'm told that anyone who knows anything about chemistry knows this is a big deal and the sections will fade. Every lab I've worked in used xylene to clear frozen sections. I understand that I messed up because this isn't our proceedure, but I don't believe I sacrificed patient care. Am I wrong? Lynn From: lynn...@hotmail.com To: histonet@lists.utsouthwestern.edu Date: Thu, 18 Jun 2009 19:47:14 -0500 Subject: [Histonet] xylene and frozen sections Lynn _ Insert movie times and more without leaving Hotmail®. http://windowslive.com/Tutorial/Hotmail/QuickAdd?ocid=TXT_TAGLM_WL_HM_Tutorial_QuickAdd_062009 Insert movie times and more without leaving Hotmail®. See how. _ Hotmail® has ever-growing storage! Don’t worry about storage limits. http://windowslive.com/Tutorial/Hotmail/Storage?ocid=TXT_TAGLM_WL_HM_Tutorial_Storage_062009___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
