From Tanni.Ahmed <@t> intervet.com Wed Feb 1 03:28:45 2006 From: Tanni.Ahmed <@t> intervet.com (Ahmed, T (Tanni)) Date: Wed Feb 1 03:30:05 2006 Subject: [Histonet] Re: Immunohistochemistry on Poultry Tissues Message-ID: <11DBF8AFF1DC0F4A9B51A23F8E5BDA332B32DF@mksn83.d50.intra> Dear Marilyn, We have recently been working on poultry tissue and performed IHC to detect Infectious Bursal Disease Virus using the Vector Elite ABC kit - works wonderfully says our pathologist! I am not sure you can test using this kit for Adeno virus but it may be worth a go. Regards, Tanni Tanni S Ahmed Scientific Officer - Histopathology, R&D Intervet UK Ltd. Walton Manor, Milton Keynes, Buckinghamshire, UK. E-mail: tanni.ahmed@intervet.com Message: 12 Date: Mon, 30 Jan 2006 20:08:39 -0700 From: "Marilyn Johnson" Subject: [Histonet] Immunohistochemistry on Poultry Tissues To: Message-ID: <001801c62613$a2286bf0$6401a8c0@VALUED20606295> Content-Type: text/plain; charset="iso-8859-1" Hi Histonetters, If anyone performs IHC on poultry tissues please give me a list of your tests. I am particularily interested in testing for Adeno virus in turkeys. Thanks in advance to any replies to my request. Marilyn Johnson Alberta Agriculture Edmonton, Alberta, Canada, -------------------------------------- This message, including attachments, is confidential and may be privileged. If you are not an intended recipient, please notify the sender then delete and destroy the original message and all copies. You should not copy, forward and/or disclose this message, in whole or in part, without permission of the sender. -------------------------------------- From kemlo.rogerson <@t> waht.swest.nhs.uk Wed Feb 1 03:38:43 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Wed Feb 1 03:38:59 2006 Subject: [Histonet] processing Message-ID: Think rodent tissue can process on the 'hard' side. Wonder what makes it 'hard'? Is it that the bonds between the proteins are 'stronger' or the proteins are 'nearer' together? I mean if we could figure out what made tissue 'hard' molecularly then we'd know which the culprit in the processing was. Sorry to ramble. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Steven Coakley [mailto:sjchtascp@yahoo.com] Sent: Tuesday, January 31, 2006 6:56 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] processing Good afternoon, I'm working with mouse/rat muscle and trying to fine tune my processing schecdule. My last run of muscle were very dry. I have a copy of the NSH Animal Processing Manual and trying to strike a balance between those listed on page 5-6. I've noticed most of the protocols do not call for heat or P/V. The scedule I currently use is as follows: 1-70% (hold), 1-80%, 2-95%, 3-100%, 3-xylene all for 30 minutes each and at 38C with P/V. 3 paraffins, 45 minutes each at 55C with P/V. I'm fairly new to research and still trying to make the "mental" switch in handling the different tissue. Thanks everyone, Steve --------------------------------- Bring words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From andromeda_tm <@t> libero.it Wed Feb 1 04:45:37 2006 From: andromeda_tm <@t> libero.it (Massimo) Date: Wed Feb 1 04:45:47 2006 Subject: [Histonet] Aceto Carmine ... Message-ID: Hi everyone, would any of you give me some information about the procedure on using Aceto Carmine for nucleos staining and what can I use like contrast colour? Thank you in advance for your assistance. Best Regards, Massimo From Histology <@t> acute.sney.nhs.uk Wed Feb 1 05:36:32 2006 From: Histology <@t> acute.sney.nhs.uk (Histology (Path Lab)) Date: Wed Feb 1 05:35:45 2006 Subject: [Histonet] Dako VEGF Message-ID: <4EAF6D4C18D9384492E6395AF94D81CB04E2C934@liam> Hi, Im doing an MSc research project but am pretty strapped for antibody and don't have enough to do a thorough optimization. Im using Dako monoclonal mouse anti-human VEGF (clone VG1) on sections of colon and rectum, and would appreciate any info anyone has who uses this antibody. In particular im after info on retrieval and dilution. Dako recommends 1:25-1.50 but if anyone has a cheat (i.e high dilution with overnight incubation), it would really help me out. Ive tried 20mins microwave with high pH solution (as recommended)using tonsil on coated slides but sections came off! Does anyone hotplate sections or oven overnight (40/60degrees)? Microwave with normal pH buffer wasn't too impressive! What tissue do you use as a control? Please help im on a tiny budget and need to get this up and running using as little as possible! Cheers. From jwatson <@t> gnf.org Wed Feb 1 10:04:19 2006 From: jwatson <@t> gnf.org (James Watson) Date: Wed Feb 1 10:04:34 2006 Subject: [Histonet] processing References: <20060131185548.50263.qmail@web90210.mail.scd.yahoo.com> Message-ID: Steve, I routinely use 5% glycerin in the absolute alcohols for my mouse tissues. this has shortened our soaking times and eliminated the drying artifact. I started using this years ago on whole cross sections of dog hearts that were processed at a 2.5 cm thickness and on frog leg muscle that was very dry. I am in the process of completeing some comparative studies and hope to get this published soon. if you want my processing schedules let me know. James Watson GNF San Diego jwatson@gnf.org ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Steven Coakley Sent: Tue 1/31/2006 10:55 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] processing Good afternoon, I'm working with mouse/rat muscle and trying to fine tune my processing schecdule. My last run of muscle were very dry. I have a copy of the NSH Animal Processing Manual and trying to strike a balance between those listed on page 5-6. I've noticed most of the protocols do not call for heat or P/V. The scedule I currently use is as follows: 1-70% (hold), 1-80%, 2-95%, 3-100%, 3-xylene all for 30 minutes each and at 38C with P/V. 3 paraffins, 45 minutes each at 55C with P/V. I'm fairly new to research and still trying to make the "mental" switch in handling the different tissue. Thanks everyone, Steve --------------------------------- Bring words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cforster <@t> umn.edu Wed Feb 1 10:29:35 2006 From: cforster <@t> umn.edu (cforster) Date: Wed Feb 1 10:29:45 2006 Subject: [Histonet] Tri-State Spring Meeting-MN Message-ID: <200602011629.k11GTZ9u010599@vanguard.software.umn.edu> Hello to all in Histoland, On behalf of the Minnesota Society for Histotechnology I would like to extend an invitation to each one of you. The MN/IA/WI Tri-State Spring Symposium is coming in April.We have an exciting program prepared. Look over the details and come and join us. If you have any questions please feel free to contact one of us. Sincerely, Colleen Forster MSH President Subject: 2006 Tri-State Spring Symposium - April 19th-21st - Madison, Wisconsin Date: 1Feb06 5:38am 2006 Tri-State Spring Symposium Sponsored by: Minnesota Society for Histotechnologists Wisconsin Histology Society Iowa Society of Histotechnology Dates: April 19-21, 2006 Meeting Location: The Madison Concourse Hotel 1 West Dayton Street Madison, Wisconsin Phone: (800) 356-8293 www.concoursehotel.com Room Rates: Wednesday night (April 19) $89.00* Thursday night (April 20) $89.00* Free underground parking for hotel guests *Standard guest room plus tax Check In/Check Out Time Check In Time – 3:00 p.m. Check Out Time – 11:00 a.m. Hotel Reservation Information: Room reservations should be made directly with the Madison Concourse Hotel at 1-800-356-8293 To secure symposium rates, make your reservations early and indicate that you are affiliated with the Tri-State Histology Symposium. All reservations must be guaranteed with a major credit card. Reservation deadline for the special Tri-State Symposium room rate is March 21, 2006. Directions to the Madison Concourse Hotel www.mapquest.com www.concoursehotel.com (maps & directions) SCHEDULE Wednesday, April 19 7:00 – 9:00 pm Registration and Product Show Pizza Party - Courtesy of Histotronix Inc. Thursday, April 20 8:30 – 12:00 From Collection to Management; Techniques for the Management of Small Tissue Specimens James McCormick M.D., M. Lamar Jones Lee Dickey, Skip Brown, McCormick Scientific 10:00 - 10:30 Break With Exhibitors 12:00 – 1:00 Lunch With Exhibitors 1:00 – 4:30 Workshop #1: Are You For Real? Troubleshooting What Isn’t Lori Ann Cummings, Vision BioSystems Inc. 1:00 – 4:30 Workshop #2: How LEAN Principles Can Be Applied to the Histology Laboratory Janice Mahoney, Omaha, NE 1:00 – 4:30 Workshop #3: Specimen Tracking in the Histology Laboratory Bill Martin, Thermo Electron Corporation 2:30 – 3:00 Break with Exhibitors 4:45 – 5:45 WHS Business Meeting 6:00 – 7:00 Cocktail Hour Courtesy of Leica Microsystems 7:00 – 10:00 Let the 2006 Tri-State Games Begin Sports Theme Banquet (sports attire appreciated) 10:00 - ? Hospitality Party Courtesy of Surgipath Friday, April 21 8:30 – 9:15 Breast Biopsies C.K. Chang, M.D., Madison, WI 9:15 – 10:00 An Introduction to Electron Microscopy Techniques Mark Sadowski, Milwaukee, WI 10:00 – 10:30 Break With Exhibitors 10:30 – 11:15 Histology of the Exceptional and Edible Annette Gendron,DVM, Madison, WI 11:15 – 12:00 CSI or Not Jerome Geurts, Madison, WI 12:00 – 1:00 Lunch With Exhibitors 1:00 – 4:30 Workshop #4: Are You Ready to Microwave? Donna Willis, Milestone Medical 1:00 – 4:30 Workshop #5: Eye Structures and Techniques of Gross Examination of the Globe Heather Potter, M.D., Madison, WI Amol Kulkarni, M.D., Madison, WI 1:00 – 4:30 Workshop #6: Cost Accounting for the Histology Laboratory Jan Gardner, Palestine, TX Tim Morken, Lab Vision Corporation 1:00 – 4:30 Workshop #7: *Limited to 20 people* From Amygdala to Arabidopsis: the Why’s and How’s of Vibrating Blade Microtomy Jennifer Freeland, Richard-Allan Please note: All workshops are CEU approved ------- REGISTRATION FORM BELOW -------- Minnesota Society for Histotechnology - REGISTRATION FORM - Institution: Name: Address: City/State/Zip: Phone/Fax/E-mail: Social Security #: Current MHS member: Yes ____ No ____ Registrations due by April 14, 2006 Registration refunds will not be given after April 17, 2006 ------------ Two-day Registration --------------- (includes breaks, lunches, banquet ticket, and choice of one workshop each afternoon) Members: Includes MSH dues $120.00 Non-Members: Includes MSH dues $140.00 Student Fee $ 40.00 Circle workshop 1 2 3 4 5 6 7 Friday 2nd choice________ ----------- One-day Registration with Banquet ---------------- (includes breaks, lunch, banquet ticket, and choice of one afternoon workshop) Members: Includes MSH dues $70.00 Non-Members: Includes MSH dues $80.00 Student Fee $20.00 Circle workshop 1 2 3 4 5 6 7 Friday 2nd choice________ ---------- One-Day Registration without Banquet ----------- One-day Registration (without banquet ticket) Note day attending Thursday _____ Friday _____ Members: Includes MSH dues $60.00 Non-Members: Includes MSH dues $70.00 Student Fee $20.00 Circle workshop 1 2 3 4 5 6 7 Friday 2nd choice________ ----------- Thursday Night Banquet Only ----------------- Banquet Ticket $25.00_________ ----------- Unable to attend / Paying 2006 Society Dues ---------- Unable to attend, paying 2006 dues $20.00 ($5.00 student) Circle: New Renewal Student ========== Total Payment ================= Total Payment: $ ______ Method of Payment : Awaiting Funds _____________ Make Checks Payable To: Minnesota Society for Histotechnology ========================================== Mail Check and Registration Form To: Lois Rowe 226 8 1/2 Ave. NW Rochester, MN 55901 Questions-Contacts: rowe.lois@mayo.edu or Colleen Forster HT(ASCP)QIHC Anatomic Pathology Research Laboratory Department of Laboratory Medicine and Pathology B173 PWB, MMC76 420 Delaware St. SE Minneapolis, MN 55455 email: cforster@umn.edu (612)626-1930 Phone (612)626-1930 Fax (612)624-4660 Colleen Forster HT(ASCP)QIHC Divison of Neuropathology J105 Diehl Hall, MMC174 420 Delaware St. SE Minneapolis, MN 55455 612-626-0436 612-625-0440 (f) From YuJ2 <@t> upmc.edu Wed Feb 1 11:56:15 2006 From: YuJ2 <@t> upmc.edu (Yu, Jian) Date: Wed Feb 1 11:56:25 2006 Subject: [Histonet] Active caspase 3 staining of mouse small intestine Message-ID: <2554B4CA518D504A81E6908E39478A6A0D3CDF97@1upmc-msx10.isdip.upmc.edu> Thanks to several of you who have given me a lot of great suggestions on how to get good cross sections of mouse small intestine One of you mentioned that you do Caspase 3 staining routinely with the small intestine, sorry that I could not find your email anyone. I have been using the CAM1 Ab from BD, which works well for IF on frozen sections but have a lot of background for IHC. Unfortunately the frozen sections do not have the greatest structures. I would very much appreciate that if you could share your experience with paraffin sections. Thanks again! ******************************************************** Jian Yu, Ph. D. University of Pittsburgh Cancer Institute Hillman Cancer Center Research Pavilion Office suite 2.26h, Laboratory 2.43 5117 Centre Avenue, Pittsburgh, PA 15213 Phone : 412-623-7786, (Lab) 412-623-3255 Fax: 412-623-7778 Email: yuj2@upmc.edu ******************************************************** From gcallis <@t> montana.edu Wed Feb 1 12:35:14 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Feb 1 12:35:28 2006 Subject: [Histonet] processing In-Reply-To: References: Message-ID: <6.0.0.22.1.20060201111051.01b4f290@gemini.msu.montana.edu> You are not rambling. Animal tissues, particularly rodent are very lean, and if one really wants to see dry animal tissue, tissues from birds of prey (hawks), other birds and reptiles can be among the worst. We custom process our rodent tissues, brain, small, etc and have shorter schedules for mouse brain versus hamster brain, that type of thing just to avoid overdehydration. The 1 hour per station or change even with 3 X 95% and 3 x 100% never seems to bother human species tissues much, but with rodent tissue, it removes too much of the bound water. some larger animals with bigger samples in a cassette seem to be ok with standard processing schedules but never with heat added. A curious thing observed over a long career in histowork, I saw the addition of heat to processing in automated processor a curious thing. When we used the older carousel (Sp?) models i.e Technicon, heat couldn't be added and processing was done at RT. True, heat may speed up the solvent exchange but it also can add to drying of tissues. I have always wondered why this has become a standard method over the years other than trying to speed up processing, but always felt alternating vacuum and pressure to be the best way to do that along with paying attention to length of processing schedules. What one needs to do is find the correct balance so that the free water in the tissue spaces is removed and not the bound water on protein molecules. Overdehydration and adding heat to processing will exacerbate the removal of bound water. Consequently, heat is never added to our animal tissue processing steps. At 02:38 AM 2/1/2006, you wrote: >Think rodent tissue can process on the 'hard' side. Wonder what makes it >'hard'? Is it that the bonds between the proteins are 'stronger' or the >proteins are 'nearer' together? I mean if we could figure out what made >tissue 'hard' molecularly then we'd know which the culprit in the processing >was. > >Sorry to ramble. > >Kemlo Rogerson >Pathology Manager >Ext 3311 >DD 01934 647057 >Mob 07749 754194 > > > > >-----Original Message----- >From: Steven Coakley [mailto:sjchtascp@yahoo.com] >Sent: Tuesday, January 31, 2006 6:56 PM >To: Histonet@lists.utsouthwestern.edu >Subject: [Histonet] processing > >Good afternoon, > > I'm working with mouse/rat muscle and trying to fine tune my processing >schecdule. > My last run of muscle were very dry. I have a copy of the NSH Animal >Processing Manual and trying to strike a balance between those listed on >page 5-6. I've noticed most of the protocols do not call for heat or P/V. >The scedule I currently use is as follows: 1-70% (hold), 1-80%, 2-95%, >3-100%, 3-xylene all for 30 minutes each and at 38C with P/V. 3 paraffins, >45 minutes each at 55C with P/V. I'm fairly new to research and still >trying to make the "mental" switch in handling the different tissue. > > Thanks everyone, > > Steve > > > >--------------------------------- >Bring words and photos together (easily) with > PhotoMail - it's free and works with Yahoo! Mail. >_______________________________________________ >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 Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From cbass <@t> bidmc.harvard.edu Wed Feb 1 12:52:03 2006 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Wed Feb 1 12:51:43 2006 Subject: [Histonet] collecting fresh tissue prior to perfusion Message-ID: Hello, I have what may be a silly question but I thought maybe it is worth a try. I am in a situation where the best way to collect liver tissue for my purposes is to perfuse the mouse with PFA. However, on occasion I need a small bit of fresh liver tissue to analyze for DNA. I was wondering if it is possible to collect a portion of a liver lobe at some point in the perfusion process before I use fixative? For example, I could start the perfusion with heparinized saline, clip a quarter of one lobe of the liver off and then switch over to the PFA. I would then proceed with the perfusion as normal. The only complication I can think of is that the loss of a significant portion of the liver could effect how well the rest of the liver perfuses. Has anyone tried this or is this something I should avoid? I am trying to cut down on the number of mice I am using. My goal is to obtain a completely fixed liver lobe that I can section all the way through. It is my understanding that the mouse liver is too large to fix by immersion in PFA, so it either has to be cut into blocks or the mouse perfused. Thanks, Caroline From kamaid <@t> iibce.edu.uy Wed Feb 1 12:55:36 2006 From: kamaid <@t> iibce.edu.uy (andres) Date: Wed Feb 1 12:55:52 2006 Subject: [Histonet] is Methyl Green suitable for water-based mounting media? In-Reply-To: References: <20060131185548.50263.qmail@web90210.mail.scd.yahoo.com> Message-ID: <6.2.1.2.0.20060201194542.02840978@localhost> Hi everybody, I'm trying to counterstain my in situ hybridizations with methyl green and want to mount them in glycerol or similar. I'd like to ask if any of you have done (or if it washes away with time...since all the protocols I've seen are followed by dehydration prior to mounting). Also, since I can't get here the already prepared solution and I've seen several ways to prepare the staining solution from powder(Aldrich) any suggestion about how to do it?? Thanks a lot ! Andr?s Kamaid Dpto. Histolog?a Facultad de Medicina Universidad de la Rep?blica Montevideo - Uruguay At 17:04 01/02/2006, James Watson wrote: >Steve, > >I routinely use 5% glycerin in the absolute alcohols for my mouse >tissues. this has shortened our soaking times and eliminated the drying >artifact. I started using this years ago on whole cross sections of dog >hearts that were processed at a 2.5 cm thickness and on frog leg muscle >that was very dry. I am in the process of completeing some comparative >studies and hope to get this published soon. if you want my processing >schedules let me know. > >James Watson >GNF San Diego >jwatson@gnf.org > >________________________________ > >From: histonet-bounces@lists.utsouthwestern.edu on behalf of Steven Coakley >Sent: Tue 1/31/2006 10:55 AM >To: Histonet@lists.utsouthwestern.edu >Subject: [Histonet] processing > > > >Good afternoon, > > I'm working with mouse/rat muscle and trying to fine tune my processing > schecdule. > My last run of muscle were very dry. I have a copy of the NSH Animal > Processing Manual and trying to strike a balance between those listed on > page 5-6. I've noticed most of the protocols do not call for heat or > P/V. The scedule I currently use is as follows: 1-70% (hold), 1-80%, > 2-95%, 3-100%, 3-xylene all for 30 minutes each and at 38C with P/V. 3 > paraffins, 45 minutes each at 55C with P/V. I'm fairly new to research > and still trying to make the "mental" switch in handling the different tissue. > > Thanks everyone, > > Steve > > > >--------------------------------- >Bring words and photos together (easily) with > PhotoMail - it's free and works with Yahoo! Mail. >_______________________________________________ >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 From ROrr <@t> enh.org Wed Feb 1 13:06:41 2006 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Wed Feb 1 13:06:49 2006 Subject: [Histonet] Hospitals that use Temps Message-ID: Hi Everyone, We are considering using a temp agency to cover some short leaves and maybe fill in PRN. What do you do with training and competency that needs to be done to fulfill JCAHo? What about the in-house hospital training? Do you adjust your training to accommodate these temps? In your experience, are the temp agencies responsible for some of this training? We are thinking a temp would need the same training as our staff. Does anyone have experience they would like to share? I'd appreciate it. Thank you Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 From Tiffany.L.Sheffield <@t> uth.tmc.edu Wed Feb 1 13:29:03 2006 From: Tiffany.L.Sheffield <@t> uth.tmc.edu (Sheffield, Tiffany L) Date: Wed Feb 1 13:29:08 2006 Subject: [Histonet] Bone/ Cartilage Interface Message-ID: Quick question- Has anyone done any Ab staining of the bone cartilage interface? If you have and can share the type of Ab and/or tips it would be greatly appreciated. Thanks! Tiffany Sheffield-Lopez, HT(ASCP) Supervisor, Bone Histomorphometry & Biomaterials Lab Department of Orthopaedic Surgery UTHSC-Houston Medical School 6431 Fannin, Suite 6.144 MSB Houston , TX 77030 713-500-6803 WK 713-500-0729 Fax From tkngflght <@t> yahoo.com Wed Feb 1 13:49:25 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Wed Feb 1 13:49:34 2006 Subject: [Histonet] Hospitals that use Temps In-Reply-To: Message-ID: <20060201194925.21945.qmail@web50907.mail.yahoo.com> Hi Becky- I operate a Histo/Path Anatomy placement agency (permanent staffing only but have done temp services) so let's see if I can help you with your current situation: You are obligated to have them trained in all the things JCAHO requires (all those for which you train your perm employess) and you need to have a file for each one in the same place you keep your regular employee records. There are different ways to get there. Some of the bigger agencies conduct this training by video and online reading when they contract the tech. Talk to your recruiting contact at each agency. Some of these people have been at other facilities as permanents and can provide proof from their own records or by requesting copies from their last employer. Rarely an ex-employer won't share these records putting you back at square one. Be careful with dates if these are annual training situations like safety and BBPathogens. Most facilities that regularly use temps bring them in on the hiring cycle and put them through a full orientation. You DO pay for the tech's time but it's far more cost effective than a big hit from JCAHO or any of the other governing bodies requiring safety, BBPathogens, etc. If your HR has a mini-orientation that allows an employee to start 'off-cycle' but covers all the bases then that is the best way to cover all your requirements in the most cost-effective manner. If your HR contact isn't sure and you have a nurse recruiter in your facility, this person will probably know in one phone call what you need to do to get your new temps up to speed. Please consider that for some of this, training is site specific, and the care with which you orient these folks will come back to you in many ways as they will be better temps knowing you care enough to train them correctly and well. I temped on and off for over ten years and there are labs where I stayed as a permanent because I was well treated and loved the job. Most temps are consciencious professionals and great workers. I hope you've been staffed with several of these as they really contribute and get you through the shortage with grace. Please let me know if you are seeking permanent solutions in your facility. Our services are a fraction of the other agencies--I have a regular job and helping good techs and great labs find each other is a 'calling' -- I've love to help. Cheryl Kerry Full Staff Inc 281.883.7704 ----- Original Message ---- From: "Orr, Rebecca" To: histonet@lists.utsouthwestern.edu Sent: Wednesday, February 01, 2006 1:06:41 PM Subject: [Histonet] Hospitals that use Temps Hi Everyone, We are considering using a temp agency to cover some short leaves and maybe fill in PRN. What do you do with training and competency that needs to be done to fulfill JCAHo? What about the in-house hospital training? Do you adjust your training to accommodate these temps? In your experience, are the temp agencies responsible for some of this training? We are thinking a temp would need the same training as our staff. Does anyone have experience they would like to share? I'd appreciate it. Thank you Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Karen.Heckford <@t> CHW.edu Wed Feb 1 13:53:07 2006 From: Karen.Heckford <@t> CHW.edu (Heckford, Karen - SMMC-SF) Date: Wed Feb 1 13:53:56 2006 Subject: [Histonet] IHC Help! Message-ID: Dear Histonetters, One of my pathologists needs a antibody that will show microglandular adenosis in human breast tissue. I am working with FFPE tissue and the Dako Autostainer. I have tried Neomarkers Collagen IV Ab-3 with Dako's Envision system. At the time I did not have any Protease so I used Protinease K and later used the Protease. I did get my controls to come back positive and my negative were negative all three times, but the patient tissue did not show microglandular adenosis. We did sent this tissue out and it came back positive for microglandular adenosis. I cannot figure out what I am doing wrong or perhaps I bought the wrong antibody. I would really appreciate any help. This has been bothering me for a week now. Karen Heckford HT (ASCP) CE kheckfor@chw.edu 1-415-668-1000 ext. 6167 From JosefaNava <@t> texashealth.org Wed Feb 1 13:54:36 2006 From: JosefaNava <@t> texashealth.org (Nava, Josefa) Date: Wed Feb 1 13:54:53 2006 Subject: [Histonet] looking for Antibodies-MSH6 and PMS2 Message-ID: <2C515C1049EAF5459EFD8C9B929078A41945A8@phdex03.txhealth.org> Hello Everyone , I am using Ventana BMK/XT , can someone tell me a good source of MSH6 and PMS2 that will work on the Ventana Machine. I appreciate any information. Thank you. Josie The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From dennijc <@t> vetmed.auburn.edu Wed Feb 1 14:38:37 2006 From: dennijc <@t> vetmed.auburn.edu (John C. Dennis) Date: Wed Feb 1 14:38:54 2006 Subject: [Histonet] Active caspase 3 staining of mouse small intestine In-Reply-To: <2554B4CA518D504A81E6908E39478A6A0D3CDF97@1upmc-msx10.isdip.upmc.edu> References: <2554B4CA518D504A81E6908E39478A6A0D3CDF97@1upmc-msx10.isdip.upmc.edu> Message-ID: I'm preparing a Caspase 3 preparation today, in fact. The tissue is paraffin embedded rat ethmoturbinate. I boil the slides in 10 mM Na Citrate pH 6.0 20'. I'm using Sigma's rabbit polyclonal and Vector's DAB kit. John Carroll Dennis Anatomy, Physiology, and Pharmacology 109 Greene Hall Auburn University, AL 36849 On Wed, 1 Feb 2006, Yu, Jian wrote: > Thanks to several of you who have given me a lot of great suggestions on > how to get good cross sections of mouse small intestine One of you > mentioned that you do Caspase 3 staining routinely with the small > intestine, sorry that I could not find your email anyone. I have been > using the CAM1 Ab from BD, which works well for IF on frozen sections > but have a lot of background for IHC. Unfortunately the frozen sections > do not have the greatest structures. I would very much appreciate that > if you could share your experience with paraffin sections. > > > > Thanks again! > > ******************************************************** > > Jian Yu, Ph. D. > > University of Pittsburgh Cancer Institute > > Hillman Cancer Center Research Pavilion > > Office suite 2.26h, Laboratory 2.43 > > 5117 Centre Avenue, Pittsburgh, PA 15213 > > > > Phone : 412-623-7786, (Lab) 412-623-3255 > > Fax: 412-623-7778 > > Email: yuj2@upmc.edu > > ******************************************************** > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From aep10 <@t> cornell.edu Wed Feb 1 14:58:12 2006 From: aep10 <@t> cornell.edu (Anna Elisse Beaudin) Date: Wed Feb 1 14:58:25 2006 Subject: [Histonet] Bizarre Immunofluorescence problem in mouse brain sections Message-ID: <1506.128.253.96.73.1138827492.squirrel@webmail.cornell.edu> Dear Histonet, I am having a bizarre problem with an immunofluorescence protocol that I am really hoping someone can help me with. here are the specs: I'm doing immunofluorescence for BrdU in PFA-fixed mouse brain sections that have been collected in PBS, mounted on slides, and allowed to dry 2+ days. Once dried, slides are processed for IHC as follows: Antigen retrieval (20min in .1M citrate buffer pH6.0, 95C) 2N HCl treatment 10% normal serum block followed by 10% casein block rat primary incubation O/N at 4C 5min rinse with PBS, .1% triton X, + 2X3 min PBS Secondary is a jackson Aexa488-conjugated goat anti-rat - 45 min at room temp Rinses... coverslip The problem I am encountering is the appearance of strange fiber-like pieces on my tissue that are picking up the secondary. at 40X they look like segmented strings or fibers (almost like bacteria). I have no idea where they're coming from. I did NOT have this problem with regular HRP development, and my secondary is brand new. I have this same problem even with a rat IgG control. I am still getting good specific staining, I just need to get rid of this stringy background. Does anybody have any suggestions/ideas about what this is and how I might get rid of it? I really appreciate your help. Best, Anna Beaudin Division of Nutritional Sciences Cornell University From portera <@t> msu.edu Wed Feb 1 15:12:54 2006 From: portera <@t> msu.edu (Amy Porter) Date: Wed Feb 1 15:13:29 2006 Subject: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections Message-ID: <001a01c62774$44c5ef70$8e7a0923@HistoJJ> If your willing to share - how are people monitoring the temperature of paraffin baths on tissue processors? Are you using additional thermometers directly into the paraffin baths, or just going by the digital read out on the display for the (oven on a VIP) instrument? Just curious how everyone is dealing with this particular item. Thanks in advance for the many responses and ideas I know I will get from asking this question! Amy S. Porter, HT(ASCP) QIHC Laboratory Supervisor Michigan State University Investigative HistoPathology Laboratory Department of Physiology / Division of Human Pathology 2100 Biomedical Physical Sciences Bldg. Room #2133 East Lansing, MI 48824-3320 Phone: (517) 355-6475 ext 1480 / Fax: (517) 432-1368 Email: portera@msu.edu www.humanpathology.msu.edu From rjbuesa <@t> yahoo.com Wed Feb 1 15:22:01 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Feb 1 15:22:13 2006 Subject: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections In-Reply-To: <001a01c62774$44c5ef70$8e7a0923@HistoJJ> Message-ID: <20060201212201.1914.qmail@web61215.mail.yahoo.com> Amy: The digital thermometer in the VIP is not only very accurate but the most expeditious way of checking the temperature. Just prepare a log that include all your VIPs and record the temperature once a day. That is the way I did it, and never had problems with CAP inspectors. Hope this will help you. Ren? J. Amy Porter wrote: If your willing to share - how are people monitoring the temperature of paraffin baths on tissue processors? Are you using additional thermometers directly into the paraffin baths, or just going by the digital read out on the display for the (oven on a VIP) instrument? Just curious how everyone is dealing with this particular item. Thanks in advance for the many responses and ideas I know I will get from asking this question! Amy S. Porter, HT(ASCP) QIHC Laboratory Supervisor Michigan State University Investigative HistoPathology Laboratory Department of Physiology / Division of Human Pathology 2100 Biomedical Physical Sciences Bldg. Room #2133 East Lansing, MI 48824-3320 Phone: (517) 355-6475 ext 1480 / Fax: (517) 432-1368 Email: portera@msu.edu www.humanpathology.msu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? With a free 1 GB, there's more in store with Yahoo! Mail. From jnocito <@t> pathreflab.com Wed Feb 1 15:30:05 2006 From: jnocito <@t> pathreflab.com (Joe Nocito) Date: Wed Feb 1 15:30:23 2006 Subject: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections In-Reply-To: <001a01c62774$44c5ef70$8e7a0923@HistoJJ> Message-ID: Amy, We take the temps of each individual bath, but beware. During one of my inspections, an inspector had his own thermometer and tested the temps. Although we had the temps at 60C, the baths read 65C. This was out of our range and he wrote me up. I called Sakura tech rep and had them explain to the inspector why the temp difference. Tech support told the inspector that the baths were hotter to account for the paraffin cooling off during transfer to the retort chamber. He bought it. And people wonder why I have a negative attitude about CAP. How many people have been inspected by an inspector who carries his/her own thermometer? I should have asked him to show me documentation that it was calibrated. Joe "The Toe" Nocito -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Porter Sent: Wednesday, February 01, 2006 3:13 PM To: Histonet Subject: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections If your willing to share - how are people monitoring the temperature of paraffin baths on tissue processors? Are you using additional thermometers directly into the paraffin baths, or just going by the digital read out on the display for the (oven on a VIP) instrument? Just curious how everyone is dealing with this particular item. Thanks in advance for the many responses and ideas I know I will get from asking this question! Amy S. Porter, HT(ASCP) QIHC Laboratory Supervisor Michigan State University Investigative HistoPathology Laboratory Department of Physiology / Division of Human Pathology 2100 Biomedical Physical Sciences Bldg. Room #2133 East Lansing, MI 48824-3320 Phone: (517) 355-6475 ext 1480 / Fax: (517) 432-1368 Email: portera@msu.edu www.humanpathology.msu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ander093 <@t> tc.umn.edu Wed Feb 1 15:35:54 2006 From: ander093 <@t> tc.umn.edu (LuAnn Anderson) Date: Wed Feb 1 15:36:12 2006 Subject: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections In-Reply-To: <20060201212201.1914.qmail@web61215.mail.yahoo.com> References: <001a01c62774$44c5ef70$8e7a0923@HistoJJ> <20060201212201.1914.qmail@web61215.mail.yahoo.com> Message-ID: <6.2.3.4.0.20060201153334.01d663e0@ander093.email.umn.edu> I do remember that you are required to use a separate thermometer which has been calibrated against a thermostandard and that temps from the digital readouts were not to be used. If you'd like, I can try to find where I read that. LuAnn At 03:22 PM 2/1/2006, Rene J Buesa wrote: >Amy: > The digital thermometer in the VIP is not > only very accurate but the most expeditious way of checking the temperature. > Just prepare a log that include all your VIPs > and record the temperature once a day. > That is the way I did it, and never had problems with CAP inspectors. > Hope this will help you. > Ren? J. > >Amy Porter wrote: > If your willing to share - how are people > monitoring the temperature of paraffin baths on > tissue processors? Are you using additional > thermometers directly into the paraffin baths, > or just going by the digital read out on the > display for the (oven on a VIP) instrument? > Just curious how everyone is dealing with this > particular item. Thanks in advance for the many > responses and ideas I know I will get from asking this question! >Amy S. Porter, HT(ASCP) QIHC >Laboratory Supervisor >Michigan State University >Investigative HistoPathology Laboratory >Department of Physiology / Division of Human Pathology >2100 Biomedical Physical Sciences Bldg. Room #2133 >East Lansing, MI 48824-3320 >Phone: (517) 355-6475 ext 1480 / Fax: (517) 432-1368 >Email: portera@msu.edu >www.humanpathology.msu.edu > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > >--------------------------------- >Do you Yahoo!? > With a free 1 GB, there's more in store with Yahoo! Mail. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tgoodpas <@t> fhcrc.org Wed Feb 1 17:03:56 2006 From: tgoodpas <@t> fhcrc.org (Goodpaster, Tracy A) Date: Wed Feb 1 17:04:06 2006 Subject: [Histonet] Active caspase 3 staining of mouse small intestine Message-ID: <8BD501B158A381409FE54DB421F82FCA02E8BA64@groucho.fhcrc.org> We use the cleaved caspase-3 antibody from Biocare with good success. Previously, we used the one from Cell Signalling, but switched after a side by side comparison and cost evaluation. We steam heat formalin fixed, paraffin embedded sections for 15 minutes in Dako's pH10 antigen retrieval buffer. We then use an Avidin/biotin block and serum block (15% goat + 5% human in antibody dilutent). We incubate the antibody for 90 minutes and detect it with a biotinylated goat anti-rabbit IgG at 1:400, the Vector elite RTU ABC and Dako DAB plus. We get crisp staining on mouse, dog and human tissue. I hope this helps. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Yu, Jian Sent: Wednesday, February 01, 2006 9:56 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Active caspase 3 staining of mouse small intestine Thanks to several of you who have given me a lot of great suggestions on how to get good cross sections of mouse small intestine One of you mentioned that you do Caspase 3 staining routinely with the small intestine, sorry that I could not find your email anyone. I have been using the CAM1 Ab from BD, which works well for IF on frozen sections but have a lot of background for IHC. Unfortunately the frozen sections do not have the greatest structures. I would very much appreciate that if you could share your experience with paraffin sections. Thanks again! ******************************************************** Jian Yu, Ph. D. University of Pittsburgh Cancer Institute Hillman Cancer Center Research Pavilion Office suite 2.26h, Laboratory 2.43 5117 Centre Avenue, Pittsburgh, PA 15213 Phone : 412-623-7786, (Lab) 412-623-3255 Fax: 412-623-7778 Email: yuj2@upmc.edu ******************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From wilson_jm <@t> cimar.org Wed Feb 1 17:34:32 2006 From: wilson_jm <@t> cimar.org (Jonathan Wilson) Date: Wed Feb 1 17:34:38 2006 Subject: [Histonet] citation for negative controls Message-ID: <00b901c62788$0e10b7d0$146fab81@COBITIS> Hello, I use IHC in basic research and have been digging through the archives for information on appropriate negative controls and came across the use of irrelavent antibodies. I would like to use this information in a paper I am writing and was wondering if this is cited in a review or paper. I have Harlow and Lane's book but it doesn't mention this type of control. Thank you in advance for any help. Sincerely, Jon Jonathan Wilson (PhD) ecofisiologia CIMAR Rua dos Bragas 289 4050-123 Porto Portugal office 351 22 340 1809 lab 351 22 340 1834 fax 351 22 339 0608 From AnthonyH <@t> chw.edu.au Wed Feb 1 18:01:00 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Wed Feb 1 18:01:22 2006 Subject: [Histonet] citation for negative controls Message-ID: Jonathan, The following references might be of use: Childs (1983) J Histochem Cytochem 31 (1A):168-176 Petrusz (1983) J Histochem Cytochem 31(1A):177-179. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jonathan Wilson Sent: Thursday, 2 February 2006 10:35 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] citation for negative controls Hello, I use IHC in basic research and have been digging through the archives for information on appropriate negative controls and came across the use of irrelavent antibodies. I would like to use this information in a paper I am writing and was wondering if this is cited in a review or paper. I have Harlow and Lane's book but it doesn't mention this type of control. Thank you in advance for any help. Sincerely, Jon Jonathan Wilson (PhD) ecofisiologia CIMAR Rua dos Bragas 289 4050-123 Porto Portugal office 351 22 340 1809 lab 351 22 340 1834 fax 351 22 339 0608 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From kemlo.rogerson <@t> waht.swest.nhs.uk Thu Feb 2 02:58:33 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Thu Feb 2 02:58:53 2006 Subject: [Histonet] processing Message-ID: My point stands though. What makes certain animal tissue more susceptible to hardening? You eloquently explain how we solve the problem but I've never seen the problem defined. Surely all animal tissue is intrinsically the same? So why does it process differently? Now I'm really rambling; why does it taste differently? Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Wednesday, February 01, 2006 6:35 PM To: Kemlo Rogerson; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] processing You are not rambling. Animal tissues, particularly rodent are very lean, and if one really wants to see dry animal tissue, tissues from birds of prey (hawks), other birds and reptiles can be among the worst. We custom process our rodent tissues, brain, small, etc and have shorter schedules for mouse brain versus hamster brain, that type of thing just to avoid overdehydration. The 1 hour per station or change even with 3 X 95% and 3 x 100% never seems to bother human species tissues much, but with rodent tissue, it removes too much of the bound water. some larger animals with bigger samples in a cassette seem to be ok with standard processing schedules but never with heat added. A curious thing observed over a long career in histowork, I saw the addition of heat to processing in automated processor a curious thing. When we used the older carousel (Sp?) models i.e Technicon, heat couldn't be added and processing was done at RT. True, heat may speed up the solvent exchange but it also can add to drying of tissues. I have always wondered why this has become a standard method over the years other than trying to speed up processing, but always felt alternating vacuum and pressure to be the best way to do that along with paying attention to length of processing schedules. What one needs to do is find the correct balance so that the free water in the tissue spaces is removed and not the bound water on protein molecules. Overdehydration and adding heat to processing will exacerbate the removal of bound water. Consequently, heat is never added to our animal tissue processing steps. At 02:38 AM 2/1/2006, you wrote: >Think rodent tissue can process on the 'hard' side. Wonder what makes it >'hard'? Is it that the bonds between the proteins are 'stronger' or the >proteins are 'nearer' together? I mean if we could figure out what made >tissue 'hard' molecularly then we'd know which the culprit in the processing >was. > >Sorry to ramble. > >Kemlo Rogerson >Pathology Manager >Ext 3311 >DD 01934 647057 >Mob 07749 754194 > > > > >-----Original Message----- >From: Steven Coakley [mailto:sjchtascp@yahoo.com] >Sent: Tuesday, January 31, 2006 6:56 PM >To: Histonet@lists.utsouthwestern.edu >Subject: [Histonet] processing > >Good afternoon, > > I'm working with mouse/rat muscle and trying to fine tune my processing >schecdule. > My last run of muscle were very dry. I have a copy of the NSH Animal >Processing Manual and trying to strike a balance between those listed on >page 5-6. I've noticed most of the protocols do not call for heat or P/V. >The scedule I currently use is as follows: 1-70% (hold), 1-80%, 2-95%, >3-100%, 3-xylene all for 30 minutes each and at 38C with P/V. 3 paraffins, >45 minutes each at 55C with P/V. I'm fairly new to research and still >trying to make the "mental" switch in handling the different tissue. > > Thanks everyone, > > Steve > > > >--------------------------------- >Bring words and photos together (easily) with > PhotoMail - it's free and works with Yahoo! Mail. >_______________________________________________ >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 Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From BMolinari <@t> heart.thi.tmc.edu Thu Feb 2 05:47:20 2006 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Thu Feb 2 05:53:35 2006 Subject: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections Message-ID: Once a month I put a calibrated thermometer in one of the paraffin reservoirs. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Porter Sent: Wednesday, February 01, 2006 3:13 PM To: Histonet Subject: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections If your willing to share - how are people monitoring the temperature of paraffin baths on tissue processors? Are you using additional thermometers directly into the paraffin baths, or just going by the digital read out on the display for the (oven on a VIP) instrument? Just curious how everyone is dealing with this particular item. Thanks in advance for the many responses and ideas I know I will get from asking this question! Amy S. Porter, HT(ASCP) QIHC Laboratory Supervisor Michigan State University Investigative HistoPathology Laboratory Department of Physiology / Division of Human Pathology 2100 Biomedical Physical Sciences Bldg. Room #2133 East Lansing, MI 48824-3320 Phone: (517) 355-6475 ext 1480 / Fax: (517) 432-1368 Email: portera@msu.edu www.humanpathology.msu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Susan.Ferrigon <@t> sanofi-aventis.com Thu Feb 2 07:29:53 2006 From: Susan.Ferrigon <@t> sanofi-aventis.com (Susan.Ferrigon@sanofi-aventis.com) Date: Thu Feb 2 07:30:05 2006 Subject: [Histonet] Kuppfer cells Message-ID: Hi Can anyone suggest a marker for Kuppfer cells?? Thanks Susan ------------------------------ Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de ce courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuellement pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ------------------------------ From nadams <@t> CapeCodHealth.org Thu Feb 2 08:43:58 2006 From: nadams <@t> CapeCodHealth.org (Adams, Nancy) Date: Thu Feb 2 08:44:08 2006 Subject: [Histonet] microwave transparent containers and equipment Message-ID: <17A1862099540D458C8FE9380C2BC461C0932E@fh2xmail.fhdomain1.capecodhealth.org> Good morning Does the new CAP question ANP.28860 require documentation that the containers and equipment are, indeed, microwave transparent? Somehow I think just saying yes isn't going to be enough:-) Thanks for your thoughts. Nancy Rutledge Falmouth Hospital ************************************************************ This email and any files transmitted with it are confidential. And intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please contact the system administrator for Cape Cod Healthcare. Administrator@CapeCodHealth.org ************************************************************ From emerald_lake77 <@t> yahoo.com Thu Feb 2 09:28:05 2006 From: emerald_lake77 <@t> yahoo.com (GT Hebert) Date: Thu Feb 2 09:28:21 2006 Subject: [Histonet] Kuppfer cells In-Reply-To: Message-ID: <20060202152805.96087.qmail@web31704.mail.mud.yahoo.com> Susan, I've used F4/80. Though I was trying to stain macrophages, I used the Kupffer cells in the liver as a positive control to show that the antibody was working and they stained perfectly. Gustave Hebert Scientist II Wyeth Research Cambridge MA Susan.Ferrigon@sanofi-aventis.com wrote: Hi Can anyone suggest a marker for Kuppfer cells?? Thanks Susan ------------------------------ Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de ce courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuellement pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Autos. Looking for a sweet ride? Get pricing, reviews, & more on new and used cars. From rjbuesa <@t> yahoo.com Thu Feb 2 09:44:20 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Feb 2 09:44:30 2006 Subject: [Histonet] microwave transparent containers and equipment In-Reply-To: <17A1862099540D458C8FE9380C2BC461C0932E@fh2xmail.fhdomain1.capecodhealth.org> Message-ID: <20060202154420.27452.qmail@web61225.mail.yahoo.com> Nancy: You could show them a list with the microwaves penetration (cm) of different substances. For example: polyethyl methacrylate = 300 cm (= 3 m) Teflon = 9,000 cm (= 90 m) Paraffin wax = 15,000 (=150 m) Styrofoam= 40,000 cm (=400 m) Showing them the materials your containers are made of I think that they will accept that they are indeed transparent. Hope this will help. Ren? J. "Adams, Nancy" wrote: Good morning Does the new CAP question ANP.28860 require documentation that the containers and equipment are, indeed, microwave transparent? Somehow I think just saying yes isn't going to be enough:-) Thanks for your thoughts. Nancy Rutledge Falmouth Hospital ************************************************************ This email and any files transmitted with it are confidential. And intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please contact the system administrator for Cape Cod Healthcare. Administrator@CapeCodHealth.org ************************************************************ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? With a free 1 GB, there's more in store with Yahoo! Mail. From bruyntjes <@t> voeding.tno.nl Thu Feb 2 10:09:19 2006 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Thu Feb 2 10:09:34 2006 Subject: FW: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections Message-ID: <3B070848E7C2204F9DEB8BCFD767728004A69837@ntexch1.voeding.tno.nl> Hi Amy We use an electronic thermometer (testo 915-1) which is calibrated once or twice a year. Joost Bruijntjes TNO Quality of Life Toxicology and Applied Pharmacology Zeist Holland -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Molinari, Betsy Sent: donderdag 2 februari 2006 12:47 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections Once a month I put a calibrated thermometer in one of the paraffin reservoirs. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Porter Sent: Wednesday, February 01, 2006 3:13 PM To: Histonet Subject: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections If your willing to share - how are people monitoring the temperature of paraffin baths on tissue processors? Are you using additional thermometers directly into the paraffin baths, or just going by the digital read out on the display for the (oven on a VIP) instrument? Just curious how everyone is dealing with this particular item. Thanks in advance for the many responses and ideas I know I will get from asking this question! Amy S. Porter, HT(ASCP) QIHC Laboratory Supervisor Michigan State University Investigative HistoPathology Laboratory Department of Physiology / Division of Human Pathology 2100 Biomedical Physical Sciences Bldg. Room #2133 East Lansing, MI 48824-3320 Phone: (517) 355-6475 ext 1480 / Fax: (517) 432-1368 Email: portera@msu.edu www.humanpathology.msu.edu _______________________________________________ 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 This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From gcallis <@t> montana.edu Thu Feb 2 11:02:18 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Feb 2 11:02:39 2006 Subject: [Histonet] citation for negative controls In-Reply-To: <00b901c62788$0e10b7d0$146fab81@COBITIS> References: <00b901c62788$0e10b7d0$146fab81@COBITIS> Message-ID: <6.0.0.22.1.20060202095040.01b3ce58@gemini.msu.montana.edu> Appropriate negative controls are discussed by Boenisch, in DAKO 3rd Edition Handbook Immunochemical Staining methods, and found on the DAKO website as a free pdf download. It can be done and clinical laboratories do use irrelevant antibodies at times also. Jules Elias books also discuss this, Immunohistopathology, a pracatical approach to diagnostics, ASCP press. There is a second edition although may not be available to you. If you are working with mouse, this can be a difficult thing to find at times so that you get NO staining with an irrelevant antibody. Consequently we use what is most cited in the literature, isotype matched immunoglobulin i.e. IgG controls depending on species and immunoglobulin. They come pure, biotinylated, conjugated to many things including Alexa dyes. OR you can buy whole IgG's from Jackson (whole rat IgG contains all the isotypes, and from many different species.) One that happened some time ago, we used rat serum as a control and the reviewers balked big time. We had to REPEAT what was a very difficult, tedious immunogold staining project using isotype matched controls. Since that time, we NEVER use normal serums nor an irrelevant antibody for a negative control. Having to repeat work is not fun! At 04:34 PM 2/1/2006, you wrote: >Hello, >I use IHC in basic research and have been digging through the archives for >information on appropriate negative controls and came across the use of >irrelavent antibodies. I would like to use this information in a paper I >am writing and was wondering if this is cited in a review or paper. I have >Harlow and Lane's book but it doesn't mention this type of control. >Thank you in advance for any help. >Sincerely, >Jon > > >Jonathan Wilson (PhD) >ecofisiologia CIMAR >Rua dos Bragas 289 >4050-123 Porto Portugal >office 351 22 340 1809 >lab 351 22 340 1834 >fax 351 22 339 0608 >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From Terry.Marshall <@t> rothgen.nhs.uk Thu Feb 2 10:12:44 2006 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Thu Feb 2 11:12:59 2006 Subject: [Histonet] pictures - another way? Message-ID: http://www.pando.com/welcome.html I have just come across this site and program, which allows you to send attachments of any size, overcoming restrictions most hosts put on attachment size. The program puts the file on their website and provides a link, which lasts just 14 few days. The link can then be e-mailed. This strikes me as a good alternative way to post pictures. It does require you to have the programme, but I gather this is very small. Currently it is in beta testing and you have to register an interest and get added when and if. I have just done so, and suggest a few others do the same (not all at once) then we can give it a go. Will keep you posted. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk From celebrej <@t> HHSC.CA Thu Feb 2 11:21:26 2006 From: celebrej <@t> HHSC.CA (Celebre Julia) Date: Thu Feb 2 11:21:37 2006 Subject: [Histonet] Re-processing Message-ID: <0B3BF9D810C7094FA34B17DD145C3D5E0FA825@ipemail01.hhsc.ca> Calling all processing and reprocessing experts out there.... I'm sure all labs over the years have run into processing troubles ranging from instrument malfunctions during the night to misplacement of solutions (usually a water before the xylene), causing pure horror for the technologist who opens up the retort in the morning. It's sad to say that our department is experienced in these situations, and everytime something happens to the processing it's always difficult to decide how best to reprocess the sections. Each time we've reprocessed our sections (usually due to the introduction of water before the xylene or paraffin stations) on shortened absolute alcohol, xylenes and paraffin times, they'v ended up being extremely hard and brittle, which does not make our pathologist happy.... Is there a way of reprocessing tissue without damaging it? Would adding a softener to the alcohols reduce the hardening affects of reprocessing? How much time can be safely shaved off if the tissues have already been through a regular program? Any help is greatly appreciated, Julia Celebre MLT Anatomic Pathology Hamilton General Hospital Hamilton, Ontario Canada 905-527-0271 ext 46179 email: celebrej@hhsc.ca This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From sjchtascp <@t> yahoo.com Thu Feb 2 11:22:41 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Thu Feb 2 11:22:50 2006 Subject: [Histonet] paraffin "stuff"??? Message-ID: <20060202172241.56660.qmail@web90207.mail.scd.yahoo.com> I've noticed what appears to be this off colored "fuzzy" stuff floating in the paraffin thats in my embedding dispencer. I drained the dispencer and replaced it with fresh but there it is again. Is this anything to be concerned about. "???" maybe i should check the processors paraffins. Never noticed this "stuff" before. --------------------------------- What are the most popular cars? Find out at Yahoo! Autos From pmarcum <@t> vet.upenn.edu Thu Feb 2 11:32:25 2006 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Thu Feb 2 11:32:41 2006 Subject: [Histonet] paraffin "stuff"??? In-Reply-To: <20060202172241.56660.qmail@web90207.mail.scd.yahoo.com> References: <20060202172241.56660.qmail@web90207.mail.scd.yahoo.com> Message-ID: <6.1.1.1.2.20060202123032.019c0e78@mail.vet.upenn.edu> At 12:22 PM 2/2/2006, Steven Coakley wrote: >I've noticed what appears to be this off colored "fuzzy" stuff floating >in the paraffin thats in my embedding dispencer. I drained the dispencer >and replaced it with fresh but there it is again. Is this anything to be >concerned about. "???" maybe i should check the processors >paraffins. Never noticed this "stuff" before. > > >--------------------------------- > > What are the most popular cars? Find out at Yahoo! Autos >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Hi Steve, First which paraffin are you using? Does the stuff go away when you stir it? Paraffins today all have additives that can seperate and should be stirred daily to assure this is not happening or is re mixed. Some paraffins have more additive than others and will precipitate more. Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From vd38 <@t> georgetown.edu Thu Feb 2 11:28:05 2006 From: vd38 <@t> georgetown.edu (Vernon Dailey) Date: Thu Feb 2 11:33:09 2006 Subject: [Histonet] Dopamine receptor and transporter IHC Message-ID: <43E24125.7000408@georgetown.edu> Hi Histonet, I am interested in looking at Dopamine Receptor 1 and 2 as well as Dopamine Transporter (DAT) in frozen rat brain tissue. Can anyone recommend a vendor and catalog number? Also, if possible include images/references as my PI will surely ask. Thanks in advance, -Vernon From Jackie.O'Connor <@t> abbott.com Thu Feb 2 11:35:50 2006 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Thu Feb 2 11:36:19 2006 Subject: [Histonet] paraffin "stuff"??? Message-ID: Probably impurities from the manufacturer in the paraffin - dust, dirt - odd insect parts, like we get in processed food. Steven Coakley Sent by: histonet-bounces@lists.utsouthwestern.edu 02/02/2006 11:22 AM To: Histonet@lists.utsouthwestern.edu cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] paraffin "stuff"??? I've noticed what appears to be this off colored "fuzzy" stuff floating in the paraffin thats in my embedding dispencer. I drained the dispencer and replaced it with fresh but there it is again. Is this anything to be concerned about. "???" maybe i should check the processors paraffins. Never noticed this "stuff" before. --------------------------------- What are the most popular cars? Find out at Yahoo! Autos _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Karen.Heckford <@t> CHW.edu Thu Feb 2 11:39:57 2006 From: Karen.Heckford <@t> CHW.edu (Heckford, Karen - SMMC-SF) Date: Thu Feb 2 11:49:31 2006 Subject: [Histonet] HistoQip Message-ID: Hi Everyone, I was wondering if someone could either email me or call me I need to know more information about HistoQip. I have never participated in this before and my supervisor and pathologists are leaving me in the dark. kheckfor@chw.edu or 1-415-668-1000 ext. 6167 I am in the lab between 4:30am-1pm Pacific Time Monday- Friday. Thanks in advance for your help, Karen Heckford HT (ASCP) From PMonfils <@t> Lifespan.org Thu Feb 2 12:00:41 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Thu Feb 2 12:00:53 2006 Subject: [Histonet] pictures - another way? Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171765E@lsexch.lsmaster.lifespan.org> There are a number of commercial sites that allow you to post pictures for a fee. However, most servers provide subscribers with a certain amount of personal webspace, and the necessary tools to upload documents to that webspace. I have been doing that for years, with pictures I want to make accessible to others. For example: http://members.cox.net/paulcyp/asper.jpg From DELLAV <@t> musc.edu Thu Feb 2 13:30:52 2006 From: DELLAV <@t> musc.edu (Vinnie Della Speranza) Date: Thu Feb 2 13:32:32 2006 Subject: [Histonet] HistoQip Message-ID: Karen, you can find out all you'd like to know by going to www.cap.org and at the top of the screen do a site search for HistoQIP you will get several hits that will give you details of the program Vinnie Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> "Heckford, Karen - SMMC-SF" 02/02/06 12:39PM >>> Hi Everyone, I was wondering if someone could either email me or call me I need to know more information about HistoQip. I have never participated in this before and my supervisor and pathologists are leaving me in the dark. kheckfor@chw.edu or 1-415-668-1000 ext. 6167 I am in the lab between 4:30am-1pm Pacific Time Monday- Friday. Thanks in advance for your help, Karen Heckford HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hej01 <@t> health.state.ny.us Thu Feb 2 13:52:09 2006 From: hej01 <@t> health.state.ny.us (Helen E Johnson) Date: Thu Feb 2 13:52:29 2006 Subject: [Histonet] WNV Immunohistochemistry Message-ID: Hi Histonetters, Is there anyone out there that has had success doing WNV Immunohistochemistry on FFPE mouse tissue? If so, could you please share your protocol and vendors. Thanks. Helen e-mail hej01@health.state.ny.us From Janet.Bonner <@t> FLHOSP.ORG Thu Feb 2 14:14:45 2006 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Thu Feb 2 14:16:20 2006 Subject: [Histonet] paraffin "stuff"??? Message-ID: <2813096D35FBB54FBE3EB3F1991024D60CDF8E@fhosxchmb003.ADVENTISTCORP.NET> Check the temperature of the paraffin. It could be bits of the plastic component not melting in solution - just existing on a borderline temperature. -Janet ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Steven Coakley Sent: Thu 2/2/2006 12:22 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] paraffin "stuff"??? I've noticed what appears to be this off colored "fuzzy" stuff floating in the paraffin thats in my embedding dispencer. I drained the dispencer and replaced it with fresh but there it is again. Is this anything to be concerned about. "???" maybe i should check the processors paraffins. Never noticed this "stuff" before. --------------------------------- What are the most popular cars? Find out at Yahoo! Autos _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Thu Feb 2 14:30:05 2006 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Thu Feb 2 14:30:34 2006 Subject: [Histonet] paraffin "stuff"??? Message-ID: There always seems to be some kind of dirt in paraffin, no matter who the manufacturer is - it's not created in a sterile environment by any means. Dirt or dust is bound to get in during the manufacturing/packaging process. (Did you ever check to see how many insect parts are allowed by the FDA in a jar of baby applesauce?) Anyway, you've got a screen filter in the bottom of your embedding center to filter out the chunks, right? There is always some "dark stuff" when you wipe that tank out from time to time, right? Dirt. If it gets in your block during embedding, you'll know by the unexplained scratches in your sections. I've never known this cosmic dust to create any real problems during processing/infiltration - sometimes the gunk you wipe out of the infiltration pots are carryover from all the processing, little collections of tissue debris and the like. Just keep your equipment clean and wipe that stuff out when you see it - and check out the stuff about bugs in baby food, that's WAY more interesting. All processed food is allowed a certain amount of foreign matter before it's rejected - and that IS supposed to be a much cleaner manufacturing process. The moral of the story - grow your own. (I don't - my mantra is whatever doesn't kill us makes us stronger.) Jackie O' "Bonner, Janet" Sent by: histonet-bounces@lists.utsouthwestern.edu 02/02/2006 02:14 PM To: "Steven Coakley" , Histonet@lists.utsouthwestern.edu cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: RE: [Histonet] paraffin "stuff"??? Check the temperature of the paraffin. It could be bits of the plastic component not melting in solution - just existing on a borderline temperature. -Janet ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Steven Coakley Sent: Thu 2/2/2006 12:22 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] paraffin "stuff"??? I've noticed what appears to be this off colored "fuzzy" stuff floating in the paraffin thats in my embedding dispencer. I drained the dispencer and replaced it with fresh but there it is again. Is this anything to be concerned about. "???" maybe i should check the processors paraffins. Never noticed this "stuff" before. --------------------------------- What are the most popular cars? Find out at Yahoo! Autos _______________________________________________ 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 From shive003 <@t> umn.edu Thu Feb 2 15:26:09 2006 From: shive003 <@t> umn.edu (Jan Shivers) Date: Thu Feb 2 15:26:36 2006 Subject: [Histonet] WNV Immunohistochemistry References: Message-ID: <006d01c6283f$49363da0$41065486@auxs.umn.edu> I haven't stained for WNV in mice, but I've stained for it in squirrels. Do you need to know about staining for WNV in mouse tissue specifically? Jan Shivers U of MN Vet Diag Lab ----- Original Message ----- From: "Helen E Johnson" To: Sent: Thursday, February 02, 2006 1:52 PM Subject: [Histonet] WNV Immunohistochemistry > > Hi Histonetters, > > Is there anyone out there that has had success doing WNV > Immunohistochemistry on FFPE mouse tissue? If so, could you please share > your protocol and vendors. > > > Thanks. Helen > > e-mail hej01@health.state.ny.us > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From csesvold <@t> usuhs.mil Thu Feb 2 15:33:24 2006 From: csesvold <@t> usuhs.mil (Carmen Contreras-Sesvold) Date: Thu Feb 2 15:33:57 2006 Subject: [Histonet] paraffin embedding of frozen tissue Message-ID: Greetings histonetters, I have recently found that sectioning frozen lung is a bit difficult. I was wondering if anyone has ever paraffin embedded previously fixed (and cryproserved) frozen sections? And if so what were the results? I am performing immunohistochemistry / immunofluorecent staining on mice lung tissue and I was just wondering if it was possible. I also encourge advice on the subject matter. I have learned quite a bit from this web site. Thank you very much. Carmen Contreras-Sesvold From jlinda <@t> ces.clemson.edu Thu Feb 2 15:41:19 2006 From: jlinda <@t> ces.clemson.edu (Linda Jenkins) Date: Thu Feb 2 15:38:33 2006 Subject: [Histonet] It REALLY is the last day to register for the Region III hotel! Message-ID: <5.2.1.1.2.20060202163636.00a8d970@mailhost.ces.clemson.edu> Yes...that is correct...we have finally come to the very last extension for the Region III hotel at the reduced rate. Please call 843-856-0028 to make a hotel reservation. Ask for the Region III rate. Meeting registration deadline is March 1st. Hope to see you there! Linda Linda Jenkins, HT Clemson University Dept. of Bioengineering Clemson, SC 29634-0905 864.656.5553 http://www.ces.clemson.edu/bio/research/histo.htm From rjbuesa <@t> yahoo.com Thu Feb 2 15:58:01 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Feb 2 15:58:12 2006 Subject: [Histonet] paraffin embedding of frozen tissue In-Reply-To: Message-ID: <20060202215801.87494.qmail@web61219.mail.yahoo.com> Carmen: We used to have a frozen tumors bank (tissues preserved at -80?C) and when some study was going to be done we either cut frozen sections, or processed as regular tissues. Either way you have to fix it (as if it were a "fresh" tissue) and processed it with any regular protocol suitable for the type of tissue. Now, for immunohistochemistry (IHC), since you have fixed it in formalin, you will have to perform heat induced epitope retrieval before the IHC. We used to do the same thing for the tissues left-over from the frozen sectioning diagnostic procedure. Hope this will help you. Ren? J. Carmen Contreras-Sesvold wrote: Greetings histonetters, I have recently found that sectioning frozen lung is a bit difficult. I was wondering if anyone has ever paraffin embedded previously fixed (and cryproserved) frozen sections? And if so what were the results? I am performing immunohistochemistry / immunofluorecent staining on mice lung tissue and I was just wondering if it was possible. I also encourge advice on the subject matter. I have learned quite a bit from this web site. Thank you very much. Carmen Contreras-Sesvold _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Bring words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. From AnthonyH <@t> chw.edu.au Thu Feb 2 16:23:21 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Thu Feb 2 16:25:24 2006 Subject: [Histonet] Re-processing Message-ID: The following technique from Johnson (2003) Histologic 36(1):21-22 works very well: While the tissue is still hot (ie the wax is still molten) blot the tissue dry, place it back in its cassette and place the cassette in 10% formalin for reprocessing. I have often had recourse to use this method and have found it to dramatically improve the results in at least 90% of cases. The excellent results are probably due to the protective nature of the wax present in the adequately processed portions of the block. This insulates the tissue from the harmful effects of ethanol on the adequately processed portions of the tissue preventing the tissue from becoming hard and brittle (Johnson 2003). Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Celebre Julia Sent: Friday, 3 February 2006 4:21 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Re-processing Calling all processing and reprocessing experts out there.... I'm sure all labs over the years have run into processing troubles ranging from instrument malfunctions during the night to misplacement of solutions (usually a water before the xylene), causing pure horror for the technologist who opens up the retort in the morning. It's sad to say that our department is experienced in these situations, and everytime something happens to the processing it's always difficult to decide how best to reprocess the sections. Each time we've reprocessed our sections (usually due to the introduction of water before the xylene or paraffin stations) on shortened absolute alcohol, xylenes and paraffin times, they'v ended up being extremely hard and brittle, which does not make our pathologist happy.... Is there a way of reprocessing tissue without damaging it? Would adding a softener to the alcohols reduce the hardening affects of reprocessing? How much time can be safely shaved off if the tissues have already been through a regular program? Any help is greatly appreciated, Julia Celebre MLT Anatomic Pathology Hamilton General Hospital Hamilton, Ontario Canada 905-527-0271 ext 46179 email: celebrej@hhsc.ca This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From bjdewe <@t> aol.com Thu Feb 2 16:31:45 2006 From: bjdewe <@t> aol.com (bjdewe@aol.com) Date: Thu Feb 2 16:32:04 2006 Subject: [Histonet] Alkaline Phosphatase Message-ID: <8C7F65F950FF976-59C-3AEF@mblk-d33.sysops.aol.com> Hi all, Maybe you can give me some advice. I have been staining some bone tissue for alkaline phosphatase using a Bone Alk Phos antibody. I was getting reasonable staining until recently after I started putting the tissue in a 60degree C oven to help adhere the tissue to the + slides. I was having some trouble with the sections coming off. I'm now wondering if my problem is the heat on the tissue making the AlkPhos inactive. Comments? TIA... Lorie ******************************************************************* Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s)and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ******************************************************************* From rnrkenner <@t> netscape.com Thu Feb 2 18:42:15 2006 From: rnrkenner <@t> netscape.com (rnrkenner@netscape.com) Date: Thu Feb 2 18:42:24 2006 Subject: [Histonet] ?s4 electron microscopists Message-ID: <20060202164215.23A6BEA9@dm21.mta.everyone.net> Dear histonet electron microscopists - I am currently being trained in to EM to take over for our retiring microscopist. We are building a new lab, and the administration's plan is to go digital. (I have a Zeiss 900 TEM) Are any of you out there using a digital camera on your TEM? What are the advantages and disadvantages? Is the clarity comparable to a darkroom print? The theory is that I will upload my images to my computer, then email them to the pathologists ~ with the huge amount of pixels required, won't this take an awfully long time for the pathologists to download the images? Any thoughts will be greatly appreciated. Thanks! Rita Kenner HTL (ASCP) Marshfield Clinic Marshfield, WI _____________________________________________________________ Netscape. Just the Net You Need. From RSRICHMOND <@t> aol.com Thu Feb 2 21:31:48 2006 From: RSRICHMOND <@t> aol.com (RSRICHMOND@aol.com) Date: Thu Feb 2 21:32:00 2006 Subject: [Histonet] Histobath freezing medium Message-ID: <1a6.47dcadd6.311428a4@aol.com> My laboratory uses a Histobath (from Shandon/Lipshaw or whatever they're called this year), a device that keeps a liquid coolant at a temperature of around -50 C. to freeze specimens for frozen section. The liquid coolant we use in acetone, in a space that is not vented or hooded. It seems to me that this is a significant fire hazard - and we have a CAP inspection coming up. Isopentane (2-methylbutane) would probably be an even worse choice. Is there any non-flammable liquid - perhaps a liquid Freon (halogenated hydrocarbon) that could be used in this system? The documentation makes no reference to the coolant at all. Bob Richmond Samurai Pathologist From brucea <@t> unimelb.edu.au Fri Feb 3 00:31:43 2006 From: brucea <@t> unimelb.edu.au (Bruce Abaloz) Date: Fri Feb 3 00:32:12 2006 Subject: [Histonet] HELPPPPP!! Please :) Message-ID: Dear Histonet I am looking for stains that will identify the basic contents of cultured mammary alveoli (mammospheres). Broad lipid, carbohydrate and protein stains are required to determine if different hormone combinations have an effect on milk production. Any suggestions would be wonderful. Cheers Sonia -- BRUCE ABALOZ PH:61383446282 HISTOLOGIST FAX:61383447909 DEPT.of ZOOLOGY EMAIL: brucea@unimelb.edu.au THE UNIVERSITY Of MELBOURNE. VICTORIA. AUSTRALIA 3010 Nobody Can Make You Feel Inferior Without YOUR Permission - Eleanor Roosevelt DANCE LIKE NO-ONE'S WATCHING Q: What's a specimen? A: An Italian astronaut. ******************************************************************************** This email and any attachments may contain personal information or information that is otherwise privileged, confidential or otherwise protected from disclosure/subject of copyright. Any use, disclosure or copying of any part of it is prohibited. The University does not warrant that this email or any attachments are free from viruses or defects. Please check any attachments for viruses and defects before opening them. If this email is received in error please delete it and notify us by return email. The University does not necessarily share or endorse the views expressed in this email. From barbara.bublava <@t> meduniwien.ac.at Fri Feb 3 02:03:56 2006 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Fri Feb 3 02:09:14 2006 Subject: [Histonet] deep cooling plate? References: Message-ID: <009201c62898$66547970$47a9abc1@GERICHTS9XOZZ8> Hi Does any of you have expierience with cooling/freezing plates which have a deep cooling plate so that there is a "cooling chamber" It looks like a tub or bath and there are racks so that you can put your cassettes in a vertical position not only horizontal? Is it worth the extra money? thanks in advance Barbara From Susan.Ferrigon <@t> sanofi-aventis.com Fri Feb 3 02:44:06 2006 From: Susan.Ferrigon <@t> sanofi-aventis.com (Susan.Ferrigon@sanofi-aventis.com) Date: Fri Feb 3 02:44:23 2006 Subject: [Histonet] deep cooling plate? Message-ID: Hi This sounds a great idea, do you know where to buy them from?? Thanks Susan |---------+-----------------------------------------> | | "Barbara Bublava" | | | | | | Sent by: | | | histonet-bounces@lists.utsouth| | | western.edu | | | | | | | | | 03/02/2006 08:03 | | | | |---------+-----------------------------------------> >-------------------------------------------------------------------------------------------------------------------------------| | | | To: | | cc: | | Subject: [Histonet] deep cooling plate? | >-------------------------------------------------------------------------------------------------------------------------------| Hi Does any of you have expierience with cooling/freezing plates which have a deep cooling plate so that there is a "cooling chamber" It looks like a tub or bath and there are racks so that you can put your cassettes in a vertical position not only horizontal? Is it worth the extra money? thanks in advance Barbara _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de ce courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuellement pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ------------------------------ From kemlo.rogerson <@t> waht.swest.nhs.uk Fri Feb 3 02:53:06 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Fri Feb 3 02:53:18 2006 Subject: [Histonet] Re-processing Message-ID: The hardness caused by processing is presently an intellectual battle between myself and Gayle (covert to this List). I'm presently in think mode on this. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Celebre Julia [mailto:celebrej@HHSC.CA] Sent: Thursday, February 02, 2006 5:21 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Re-processing Calling all processing and reprocessing experts out there.... I'm sure all labs over the years have run into processing troubles ranging from instrument malfunctions during the night to misplacement of solutions (usually a water before the xylene), causing pure horror for the technologist who opens up the retort in the morning. It's sad to say that our department is experienced in these situations, and everytime something happens to the processing it's always difficult to decide how best to reprocess the sections. Each time we've reprocessed our sections (usually due to the introduction of water before the xylene or paraffin stations) on shortened absolute alcohol, xylenes and paraffin times, they'v ended up being extremely hard and brittle, which does not make our pathologist happy.... Is there a way of reprocessing tissue without damaging it? Would adding a softener to the alcohols reduce the hardening affects of reprocessing? How much time can be safely shaved off if the tissues have already been through a regular program? Any help is greatly appreciated, Julia Celebre MLT Anatomic Pathology Hamilton General Hospital Hamilton, Ontario Canada 905-527-0271 ext 46179 email: celebrej@hhsc.ca This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From barbara.bublava <@t> meduniwien.ac.at Fri Feb 3 04:09:27 2006 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Fri Feb 3 04:14:24 2006 Subject: [Histonet] deep cooling plate? References: Message-ID: <00d001c628a9$ec1c4ea0$47a9abc1@GERICHTS9XOZZ8> Hi Susan, I found 3 distributor (in Europe) 1. http://www.bio-optica.it/index.html (direct link to productinformation: http://85.46.16.26/BIO_OPTICA/ENG/PRODUCTS/DATASHEET/23-PF200.pdf ) 2. www.medite.de (http://www.medite.de/downloads/Kuehlen_dts_08_08_03.pdf ) 3. http://www.weinkauf-medizintechnik.de (http://www.weinkauf-medizintechnik.de/501101953d0b1ce05/501101953d0c22d03/5 01101953d0c49a13/index.html) best regards barbara ----- Original Message ----- From: To: "Barbara Bublava" Cc: ; Sent: Friday, February 03, 2006 9:44 AM Subject: Re: [Histonet] deep cooling plate? Hi This sounds a great idea, do you know where to buy them from?? Thanks Susan |---------+-----------------------------------------> | | "Barbara Bublava" | | | | | | Sent by: | | | histonet-bounces@lists.utsouth| | | western.edu | | | | | | | | | 03/02/2006 08:03 | | | | |---------+-----------------------------------------> >--------------------------------------------------------------------------- ----------------------------------------------------| | | | To: | | cc: | | Subject: [Histonet] deep cooling plate? | >--------------------------------------------------------------------------- ----------------------------------------------------| Hi Does any of you have expierience with cooling/freezing plates which have a deep cooling plate so that there is a "cooling chamber" It looks like a tub or bath and there are racks so that you can put your cassettes in a vertical position not only horizontal? Is it worth the extra money? thanks in advance Barbara _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de ce courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuellement pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ------------------------------ From carl.bjorkhammer <@t> waht.swest.nhs.uk Fri Feb 3 04:22:41 2006 From: carl.bjorkhammer <@t> waht.swest.nhs.uk (Niclas Bjorkhammer) Date: Fri Feb 3 04:23:00 2006 Subject: [Histonet] RE: Histonet Digest, Vol 27, Issue 2 IHC Help! (Heckford, Karen - SMMC-SF) Message-ID: Hi, There are several antibodies that can aid the differentiation between microglandular adenosis and other conditions of the breast. Although Collagen IV has shown interesting results, I personally prefer the SMA or S-100, as these will highlight the BM better than with Collagen IV. Having said that, there have been discussions relating to the use of SMA , due to the extensive cross reaction of SMA with myofibroblasts, (smooth muscle myosin heavy chains and calponin). Probably more promising is the study showing the presence of an intensely positive reaction in the epithelial lining cells of the tubules in microglandular adenosis for S-100 protein. Might be worth a try. There is a good article published in 1996 that discusses the various immunohistochemical markers. The role of immunocytochemical markers in the differential diagnosis of proliferative and neoplastic lesions of the breast. Mod Pathol. 1996 Jan;9(1):57-62. Niclas Bj?rkhammer Senior Biomedical Scientist Department of Pathology, Histopathology Weston General Hospital Weston Area Health Trust 01934 647 056 ext 3313 carl.bjorkhammer@waht.swest.nhs.uk -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: None To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 27, Issue 2 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-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@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. RE: processing (Gayle Callis) 2. collecting fresh tissue prior to perfusion (Caroline Bass) 3. is Methyl Green suitable for water-based mounting media? (andres) 4. Hospitals that use Temps (Orr, Rebecca) 5. Bone/ Cartilage Interface (Sheffield, Tiffany L) 6. Re: Hospitals that use Temps (Cheryl) 7. IHC Help! (Heckford, Karen - SMMC-SF) 8. looking for Antibodies-MSH6 and PMS2 (Nava, Josefa) 9. Re: Active caspase 3 staining of mouse small intestine (John C. Dennis) 10. Bizarre Immunofluorescence problem in mouse brain sections (Anna Elisse Beaudin) 11. Tissue Processor Paraffin Temps for CAP Inspections (Amy Porter) 12. Re: Tissue Processor Paraffin Temps for CAP Inspections (Rene J Buesa) 13. RE: Tissue Processor Paraffin Temps for CAP Inspections (Joe Nocito) 14. Re: Tissue Processor Paraffin Temps for CAP Inspections (LuAnn Anderson) 15. RE: Active caspase 3 staining of mouse small intestine (Goodpaster, Tracy A) 16. citation for negative controls (Jonathan Wilson) 17. RE: citation for negative controls (Tony Henwood) 18. RE: processing (Kemlo Rogerson) 19. RE: Tissue Processor Paraffin Temps for CAP Inspections (Molinari, Betsy) 20. Kuppfer cells (Susan.Ferrigon@sanofi-aventis.com) 21. microwave transparent containers and equipment (Adams, Nancy) 22. Re: Kuppfer cells (GT Hebert) 23. Re: microwave transparent containers and equipment (Rene J Buesa) 24. FW: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections (Bruijntjes, J.P.) 25. Re: citation for negative controls (Gayle Callis) ---------------------------------------------------------------------- Message: 1 Date: Wed, 01 Feb 2006 11:35:14 -0700 From: Gayle Callis Subject: RE: [Histonet] processing To: Kemlo Rogerson , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20060201111051.01b4f290@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed You are not rambling. Animal tissues, particularly rodent are very lean, and if one really wants to see dry animal tissue, tissues from birds of prey (hawks), other birds and reptiles can be among the worst. We custom process our rodent tissues, brain, small, etc and have shorter schedules for mouse brain versus hamster brain, that type of thing just to avoid overdehydration. The 1 hour per station or change even with 3 X 95% and 3 x 100% never seems to bother human species tissues much, but with rodent tissue, it removes too much of the bound water. some larger animals with bigger samples in a cassette seem to be ok with standard processing schedules but never with heat added. A curious thing observed over a long career in histowork, I saw the addition of heat to processing in automated processor a curious thing. When we used the older carousel (Sp?) models i.e Technicon, heat couldn't be added and processing was done at RT. True, heat may speed up the solvent exchange but it also can add to drying of tissues. I have always wondered why this has become a standard method over the years other than trying to speed up processing, but always felt alternating vacuum and pressure to be the best way to do that along with paying attention to length of processing schedules. What one needs to do is find the correct balance so that the free water in the tissue spaces is removed and not the bound water on protein molecules. Overdehydration and adding heat to processing will exacerbate the removal of bound water. Consequently, heat is never added to our animal tissue processing steps. At 02:38 AM 2/1/2006, you wrote: >Think rodent tissue can process on the 'hard' side. Wonder what makes it >'hard'? Is it that the bonds between the proteins are 'stronger' or the >proteins are 'nearer' together? I mean if we could figure out what made >tissue 'hard' molecularly then we'd know which the culprit in the processing >was. > >Sorry to ramble. > >Kemlo Rogerson >Pathology Manager >Ext 3311 >DD 01934 647057 >Mob 07749 754194 > > > > >-----Original Message----- >From: Steven Coakley [mailto:sjchtascp@yahoo.com] >Sent: Tuesday, January 31, 2006 6:56 PM >To: Histonet@lists.utsouthwestern.edu >Subject: [Histonet] processing > >Good afternoon, > > I'm working with mouse/rat muscle and trying to fine tune my processing >schecdule. > My last run of muscle were very dry. I have a copy of the NSH Animal >Processing Manual and trying to strike a balance between those listed on >page 5-6. I've noticed most of the protocols do not call for heat or P/V. >The scedule I currently use is as follows: 1-70% (hold), 1-80%, 2-95%, >3-100%, 3-xylene all for 30 minutes each and at 38C with P/V. 3 paraffins, >45 minutes each at 55C with P/V. I'm fairly new to research and still >trying to make the "mental" switch in handling the different tissue. > > Thanks everyone, > > Steve > > > >--------------------------------- >Bring words and photos together (easily) with > PhotoMail - it's free and works with Yahoo! Mail. >_______________________________________________ >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 Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 2 Date: Wed, 1 Feb 2006 13:52:03 -0500 From: Caroline Bass Subject: [Histonet] collecting fresh tissue prior to perfusion To: Histonet (E-mail) Message-ID: Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed Hello, I have what may be a silly question but I thought maybe it is worth a try. I am in a situation where the best way to collect liver tissue for my purposes is to perfuse the mouse with PFA. However, on occasion I need a small bit of fresh liver tissue to analyze for DNA. I was wondering if it is possible to collect a portion of a liver lobe at some point in the perfusion process before I use fixative? For example, I could start the perfusion with heparinized saline, clip a quarter of one lobe of the liver off and then switch over to the PFA. I would then proceed with the perfusion as normal. The only complication I can think of is that the loss of a significant portion of the liver could effect how well the rest of the liver perfuses. Has anyone tried this or is this something I should avoid? I am trying to cut down on the number of mice I am using. My goal is to obtain a completely fixed liver lobe that I can section all the way through. It is my understanding that the mouse liver is too large to fix by immersion in PFA, so it either has to be cut into blocks or the mouse perfused. Thanks, Caroline ------------------------------ Message: 3 Date: Wed, 01 Feb 2006 19:55:36 +0100 From: andres Subject: [Histonet] is Methyl Green suitable for water-based mounting media? To: Message-ID: <6.2.1.2.0.20060201194542.02840978@localhost> Content-Type: text/plain; charset=iso-8859-1; format=flowed Hi everybody, I'm trying to counterstain my in situ hybridizations with methyl green and want to mount them in glycerol or similar. I'd like to ask if any of you have done (or if it washes away with time...since all the protocols I've seen are followed by dehydration prior to mounting). Also, since I can't get here the already prepared solution and I've seen several ways to prepare the staining solution from powder(Aldrich) any suggestion about how to do it?? Thanks a lot ! Andr?s Kamaid Dpto. Histolog?a Facultad de Medicina Universidad de la Rep?blica Montevideo - Uruguay At 17:04 01/02/2006, James Watson wrote: >Steve, > >I routinely use 5% glycerin in the absolute alcohols for my mouse >tissues. this has shortened our soaking times and eliminated the drying >artifact. I started using this years ago on whole cross sections of dog >hearts that were processed at a 2.5 cm thickness and on frog leg muscle >that was very dry. I am in the process of completeing some comparative >studies and hope to get this published soon. if you want my processing >schedules let me know. > >James Watson >GNF San Diego >jwatson@gnf.org > >________________________________ > >From: histonet-bounces@lists.utsouthwestern.edu on behalf of Steven Coakley >Sent: Tue 1/31/2006 10:55 AM >To: Histonet@lists.utsouthwestern.edu >Subject: [Histonet] processing > > > >Good afternoon, > > I'm working with mouse/rat muscle and trying to fine tune my processing > schecdule. > My last run of muscle were very dry. I have a copy of the NSH Animal > Processing Manual and trying to strike a balance between those listed on > page 5-6. I've noticed most of the protocols do not call for heat or > P/V. The scedule I currently use is as follows: 1-70% (hold), 1-80%, > 2-95%, 3-100%, 3-xylene all for 30 minutes each and at 38C with P/V. 3 > paraffins, 45 minutes each at 55C with P/V. I'm fairly new to research > and still trying to make the "mental" switch in handling the different tissue. > > Thanks everyone, > > Steve > > > >--------------------------------- >Bring words and photos together (easily) with > PhotoMail - it's free and works with Yahoo! Mail. >_______________________________________________ >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 ------------------------------ Message: 4 Date: Wed, 1 Feb 2006 13:06:41 -0600 From: "Orr, Rebecca" Subject: [Histonet] Hospitals that use Temps To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hi Everyone, We are considering using a temp agency to cover some short leaves and maybe fill in PRN. What do you do with training and competency that needs to be done to fulfill JCAHo? What about the in-house hospital training? Do you adjust your training to accommodate these temps? In your experience, are the temp agencies responsible for some of this training? We are thinking a temp would need the same training as our staff. Does anyone have experience they would like to share? I'd appreciate it. Thank you Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 ------------------------------ Message: 5 Date: Wed, 1 Feb 2006 13:29:03 -0600 From: "Sheffield, Tiffany L" Subject: [Histonet] Bone/ Cartilage Interface To: Message-ID: Content-Type: text/plain; charset="us-ascii" Quick question- Has anyone done any Ab staining of the bone cartilage interface? If you have and can share the type of Ab and/or tips it would be greatly appreciated. Thanks! Tiffany Sheffield-Lopez, HT(ASCP) Supervisor, Bone Histomorphometry & Biomaterials Lab Department of Orthopaedic Surgery UTHSC-Houston Medical School 6431 Fannin, Suite 6.144 MSB Houston , TX 77030 713-500-6803 WK 713-500-0729 Fax ------------------------------ Message: 6 Date: Wed, 1 Feb 2006 11:49:25 -0800 (PST) From: Cheryl Subject: Re: [Histonet] Hospitals that use Temps To: "Orr, Rebecca" , histonet@lists.utsouthwestern.edu Message-ID: <20060201194925.21945.qmail@web50907.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hi Becky- I operate a Histo/Path Anatomy placement agency (permanent staffing only but have done temp services) so let's see if I can help you with your current situation: You are obligated to have them trained in all the things JCAHO requires (all those for which you train your perm employess) and you need to have a file for each one in the same place you keep your regular employee records. There are different ways to get there. Some of the bigger agencies conduct this training by video and online reading when they contract the tech. Talk to your recruiting contact at each agency. Some of these people have been at other facilities as permanents and can provide proof from their own records or by requesting copies from their last employer. Rarely an ex-employer won't share these records putting you back at square one. Be careful with dates if these are annual training situations like safety and BBPathogens. Most facilities that regularly use temps bring them in on the hiring cycle and put them through a full orientation. You DO pay for the tech's time but it's far more cost effective than a big hit from JCAHO or any of the other governing bodies requiring safety, BBPathogens, etc. If your HR has a mini-orientation that allows an employee to start 'off-cycle' but covers all the bases then that is the best way to cover all your requirements in the most cost-effective manner. If your HR contact isn't sure and you have a nurse recruiter in your facility, this person will probably know in one phone call what you need to do to get your new temps up to speed. Please consider that for some of this, training is site specific, and the care with which you orient these folks will come back to you in many ways as they will be better temps knowing you care enough to train them correctly and well. I temped on and off for over ten years and there are labs where I stayed as a permanent because I was well treated and loved the job. Most temps are consciencious professionals and great workers. I hope you've been staffed with several of these as they really contribute and get you through the shortage with grace. Please let me know if you are seeking permanent solutions in your facility. Our services are a fraction of the other agencies--I have a regular job and helping good techs and great labs find each other is a 'calling' -- I've love to help. Cheryl Kerry Full Staff Inc 281.883.7704 ----- Original Message ---- From: "Orr, Rebecca" To: histonet@lists.utsouthwestern.edu Sent: Wednesday, February 01, 2006 1:06:41 PM Subject: [Histonet] Hospitals that use Temps Hi Everyone, We are considering using a temp agency to cover some short leaves and maybe fill in PRN. What do you do with training and competency that needs to be done to fulfill JCAHo? What about the in-house hospital training? Do you adjust your training to accommodate these temps? In your experience, are the temp agencies responsible for some of this training? We are thinking a temp would need the same training as our staff. Does anyone have experience they would like to share? I'd appreciate it. Thank you Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Wed, 1 Feb 2006 12:53:07 -0700 From: "Heckford, Karen - SMMC-SF" Subject: [Histonet] IHC Help! To: "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset=iso-8859-1 Dear Histonetters, One of my pathologists needs a antibody that will show microglandular adenosis in human breast tissue. I am working with FFPE tissue and the Dako Autostainer. I have tried Neomarkers Collagen IV Ab-3 with Dako's Envision system. At the time I did not have any Protease so I used Protinease K and later used the Protease. I did get my controls to come back positive and my negative were negative all three times, but the patient tissue did not show microglandular adenosis. We did sent this tissue out and it came back positive for microglandular adenosis. I cannot figure out what I am doing wrong or perhaps I bought the wrong antibody. I would really appreciate any help. This has been bothering me for a week now. Karen Heckford HT (ASCP) CE kheckfor@chw.edu 1-415-668-1000 ext. 6167 ------------------------------ Message: 8 Date: Wed, 1 Feb 2006 13:54:36 -0600 From: "Nava, Josefa" Subject: [Histonet] looking for Antibodies-MSH6 and PMS2 To: Message-ID: <2C515C1049EAF5459EFD8C9B929078A41945A8@phdex03.txhealth.org> Content-Type: text/plain; charset="us-ascii" Hello Everyone , I am using Ventana BMK/XT , can someone tell me a good source of MSH6 and PMS2 that will work on the Ventana Machine. I appreciate any information. Thank you. Josie The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. ------------------------------ Message: 9 Date: Wed, 1 Feb 2006 14:38:37 -0600 (CST) From: "John C. Dennis" Subject: Re: [Histonet] Active caspase 3 staining of mouse small intestine To: "Yu, Jian" Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed I'm preparing a Caspase 3 preparation today, in fact. The tissue is paraffin embedded rat ethmoturbinate. I boil the slides in 10 mM Na Citrate pH 6.0 20'. I'm using Sigma's rabbit polyclonal and Vector's DAB kit. John Carroll Dennis Anatomy, Physiology, and Pharmacology 109 Greene Hall Auburn University, AL 36849 On Wed, 1 Feb 2006, Yu, Jian wrote: > Thanks to several of you who have given me a lot of great suggestions on > how to get good cross sections of mouse small intestine One of you > mentioned that you do Caspase 3 staining routinely with the small > intestine, sorry that I could not find your email anyone. I have been > using the CAM1 Ab from BD, which works well for IF on frozen sections > but have a lot of background for IHC. Unfortunately the frozen sections > do not have the greatest structures. I would very much appreciate that > if you could share your experience with paraffin sections. > > > > Thanks again! > > ******************************************************** > > Jian Yu, Ph. D. > > University of Pittsburgh Cancer Institute > > Hillman Cancer Center Research Pavilion > > Office suite 2.26h, Laboratory 2.43 > > 5117 Centre Avenue, Pittsburgh, PA 15213 > > > > Phone : 412-623-7786, (Lab) 412-623-3255 > > Fax: 412-623-7778 > > Email: yuj2@upmc.edu > > ******************************************************** > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 10 Date: Wed, 1 Feb 2006 15:58:12 -0500 (EST) From: "Anna Elisse Beaudin" Subject: [Histonet] Bizarre Immunofluorescence problem in mouse brain sections To: histonet@lists.utsouthwestern.edu Message-ID: <1506.128.253.96.73.1138827492.squirrel@webmail.cornell.edu> Content-Type: text/plain;charset=iso-8859-1 Dear Histonet, I am having a bizarre problem with an immunofluorescence protocol that I am really hoping someone can help me with. here are the specs: I'm doing immunofluorescence for BrdU in PFA-fixed mouse brain sections that have been collected in PBS, mounted on slides, and allowed to dry 2+ days. Once dried, slides are processed for IHC as follows: Antigen retrieval (20min in .1M citrate buffer pH6.0, 95C) 2N HCl treatment 10% normal serum block followed by 10% casein block rat primary incubation O/N at 4C 5min rinse with PBS, .1% triton X, + 2X3 min PBS Secondary is a jackson Aexa488-conjugated goat anti-rat - 45 min at room temp Rinses... coverslip The problem I am encountering is the appearance of strange fiber-like pieces on my tissue that are picking up the secondary. at 40X they look like segmented strings or fibers (almost like bacteria). I have no idea where they're coming from. I did NOT have this problem with regular HRP development, and my secondary is brand new. I have this same problem even with a rat IgG control. I am still getting good specific staining, I just need to get rid of this stringy background. Does anybody have any suggestions/ideas about what this is and how I might get rid of it? I really appreciate your help. Best, Anna Beaudin Division of Nutritional Sciences Cornell University ------------------------------ Message: 11 Date: Wed, 1 Feb 2006 16:12:54 -0500 From: "Amy Porter" Subject: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections To: "Histonet" Message-ID: <001a01c62774$44c5ef70$8e7a0923@HistoJJ> Content-Type: text/plain; charset="iso-8859-1" If your willing to share - how are people monitoring the temperature of paraffin baths on tissue processors? Are you using additional thermometers directly into the paraffin baths, or just going by the digital read out on the display for the (oven on a VIP) instrument? Just curious how everyone is dealing with this particular item. Thanks in advance for the many responses and ideas I know I will get from asking this question! Amy S. Porter, HT(ASCP) QIHC Laboratory Supervisor Michigan State University Investigative HistoPathology Laboratory Department of Physiology / Division of Human Pathology 2100 Biomedical Physical Sciences Bldg. Room #2133 East Lansing, MI 48824-3320 Phone: (517) 355-6475 ext 1480 / Fax: (517) 432-1368 Email: portera@msu.edu www.humanpathology.msu.edu ------------------------------ Message: 12 Date: Wed, 1 Feb 2006 13:22:01 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections To: Amy Porter , Histonet Message-ID: <20060201212201.1914.qmail@web61215.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Amy: The digital thermometer in the VIP is not only very accurate but the most expeditious way of checking the temperature. Just prepare a log that include all your VIPs and record the temperature once a day. That is the way I did it, and never had problems with CAP inspectors. Hope this will help you. Ren? J. Amy Porter wrote: If your willing to share - how are people monitoring the temperature of paraffin baths on tissue processors? Are you using additional thermometers directly into the paraffin baths, or just going by the digital read out on the display for the (oven on a VIP) instrument? Just curious how everyone is dealing with this particular item. Thanks in advance for the many responses and ideas I know I will get from asking this question! Amy S. Porter, HT(ASCP) QIHC Laboratory Supervisor Michigan State University Investigative HistoPathology Laboratory Department of Physiology / Division of Human Pathology 2100 Biomedical Physical Sciences Bldg. Room #2133 East Lansing, MI 48824-3320 Phone: (517) 355-6475 ext 1480 / Fax: (517) 432-1368 Email: portera@msu.edu www.humanpathology.msu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? With a free 1 GB, there's more in store with Yahoo! Mail. ------------------------------ Message: 13 Date: Wed, 1 Feb 2006 15:30:05 -0600 From: "Joe Nocito" Subject: RE: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections To: "'Amy Porter'" , "'Histonet'" Message-ID: Content-Type: text/plain; charset="US-ASCII" Amy, We take the temps of each individual bath, but beware. During one of my inspections, an inspector had his own thermometer and tested the temps. Although we had the temps at 60C, the baths read 65C. This was out of our range and he wrote me up. I called Sakura tech rep and had them explain to the inspector why the temp difference. Tech support told the inspector that the baths were hotter to account for the paraffin cooling off during transfer to the retort chamber. He bought it. And people wonder why I have a negative attitude about CAP. How many people have been inspected by an inspector who carries his/her own thermometer? I should have asked him to show me documentation that it was calibrated. Joe "The Toe" Nocito -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Porter Sent: Wednesday, February 01, 2006 3:13 PM To: Histonet Subject: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections If your willing to share - how are people monitoring the temperature of paraffin baths on tissue processors? Are you using additional thermometers directly into the paraffin baths, or just going by the digital read out on the display for the (oven on a VIP) instrument? Just curious how everyone is dealing with this particular item. Thanks in advance for the many responses and ideas I know I will get from asking this question! Amy S. Porter, HT(ASCP) QIHC Laboratory Supervisor Michigan State University Investigative HistoPathology Laboratory Department of Physiology / Division of Human Pathology 2100 Biomedical Physical Sciences Bldg. Room #2133 East Lansing, MI 48824-3320 Phone: (517) 355-6475 ext 1480 / Fax: (517) 432-1368 Email: portera@msu.edu www.humanpathology.msu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 14 Date: Wed, 01 Feb 2006 15:35:54 -0600 From: LuAnn Anderson Subject: Re: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections To: Rene J Buesa , Amy Porter , Histonet Message-ID: <6.2.3.4.0.20060201153334.01d663e0@ander093.email.umn.edu> Content-Type: text/plain; charset="iso-8859-1"; format=flowed I do remember that you are required to use a separate thermometer which has been calibrated against a thermostandard and that temps from the digital readouts were not to be used. If you'd like, I can try to find where I read that. LuAnn At 03:22 PM 2/1/2006, Rene J Buesa wrote: >Amy: > The digital thermometer in the VIP is not > only very accurate but the most expeditious way of checking the temperature. > Just prepare a log that include all your VIPs > and record the temperature once a day. > That is the way I did it, and never had problems with CAP inspectors. > Hope this will help you. > Ren? J. > >Amy Porter wrote: > If your willing to share - how are people > monitoring the temperature of paraffin baths on > tissue processors? Are you using additional > thermometers directly into the paraffin baths, > or just going by the digital read out on the > display for the (oven on a VIP) instrument? > Just curious how everyone is dealing with this > particular item. Thanks in advance for the many > responses and ideas I know I will get from asking this question! >Amy S. Porter, HT(ASCP) QIHC >Laboratory Supervisor >Michigan State University >Investigative HistoPathology Laboratory >Department of Physiology / Division of Human Pathology >2100 Biomedical Physical Sciences Bldg. Room #2133 >East Lansing, MI 48824-3320 >Phone: (517) 355-6475 ext 1480 / Fax: (517) 432-1368 >Email: portera@msu.edu >www.humanpathology.msu.edu > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > >--------------------------------- >Do you Yahoo!? > With a free 1 GB, there's more in store with Yahoo! Mail. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Wed, 1 Feb 2006 15:03:56 -0800 From: "Goodpaster, Tracy A" Subject: RE: [Histonet] Active caspase 3 staining of mouse small intestine To: "Yu, Jian" , Message-ID: <8BD501B158A381409FE54DB421F82FCA02E8BA64@groucho.fhcrc.org> Content-Type: text/plain; charset="US-ASCII" We use the cleaved caspase-3 antibody from Biocare with good success. Previously, we used the one from Cell Signalling, but switched after a side by side comparison and cost evaluation. We steam heat formalin fixed, paraffin embedded sections for 15 minutes in Dako's pH10 antigen retrieval buffer. We then use an Avidin/biotin block and serum block (15% goat + 5% human in antibody dilutent). We incubate the antibody for 90 minutes and detect it with a biotinylated goat anti-rabbit IgG at 1:400, the Vector elite RTU ABC and Dako DAB plus. We get crisp staining on mouse, dog and human tissue. I hope this helps. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Yu, Jian Sent: Wednesday, February 01, 2006 9:56 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Active caspase 3 staining of mouse small intestine Thanks to several of you who have given me a lot of great suggestions on how to get good cross sections of mouse small intestine One of you mentioned that you do Caspase 3 staining routinely with the small intestine, sorry that I could not find your email anyone. I have been using the CAM1 Ab from BD, which works well for IF on frozen sections but have a lot of background for IHC. Unfortunately the frozen sections do not have the greatest structures. I would very much appreciate that if you could share your experience with paraffin sections. Thanks again! ******************************************************** Jian Yu, Ph. D. University of Pittsburgh Cancer Institute Hillman Cancer Center Research Pavilion Office suite 2.26h, Laboratory 2.43 5117 Centre Avenue, Pittsburgh, PA 15213 Phone : 412-623-7786, (Lab) 412-623-3255 Fax: 412-623-7778 Email: yuj2@upmc.edu ******************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 16 Date: Wed, 1 Feb 2006 23:34:32 -0000 From: "Jonathan Wilson" Subject: [Histonet] citation for negative controls To: Message-ID: <00b901c62788$0e10b7d0$146fab81@COBITIS> Content-Type: text/plain; charset="iso-8859-1" Hello, I use IHC in basic research and have been digging through the archives for information on appropriate negative controls and came across the use of irrelavent antibodies. I would like to use this information in a paper I am writing and was wondering if this is cited in a review or paper. I have Harlow and Lane's book but it doesn't mention this type of control. Thank you in advance for any help. Sincerely, Jon Jonathan Wilson (PhD) ecofisiologia CIMAR Rua dos Bragas 289 4050-123 Porto Portugal office 351 22 340 1809 lab 351 22 340 1834 fax 351 22 339 0608 ------------------------------ Message: 17 Date: Thu, 2 Feb 2006 11:01:00 +1100 From: "Tony Henwood" Subject: RE: [Histonet] citation for negative controls To: "Jonathan Wilson" , Message-ID: Content-Type: text/plain; charset="us-ascii" Jonathan, The following references might be of use: Childs (1983) J Histochem Cytochem 31 (1A):168-176 Petrusz (1983) J Histochem Cytochem 31(1A):177-179. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jonathan Wilson Sent: Thursday, 2 February 2006 10:35 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] citation for negative controls Hello, I use IHC in basic research and have been digging through the archives for information on appropriate negative controls and came across the use of irrelavent antibodies. I would like to use this information in a paper I am writing and was wondering if this is cited in a review or paper. I have Harlow and Lane's book but it doesn't mention this type of control. Thank you in advance for any help. Sincerely, Jon Jonathan Wilson (PhD) ecofisiologia CIMAR Rua dos Bragas 289 4050-123 Porto Portugal office 351 22 340 1809 lab 351 22 340 1834 fax 351 22 339 0608 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** ------------------------------ Message: 18 Date: Thu, 2 Feb 2006 08:58:33 -0000 From: Kemlo Rogerson Subject: RE: [Histonet] processing To: "'Gayle Callis'" , Kemlo Rogerson , Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain My point stands though. What makes certain animal tissue more susceptible to hardening? You eloquently explain how we solve the problem but I've never seen the problem defined. Surely all animal tissue is intrinsically the same? So why does it process differently? Now I'm really rambling; why does it taste differently? Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Wednesday, February 01, 2006 6:35 PM To: Kemlo Rogerson; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] processing You are not rambling. Animal tissues, particularly rodent are very lean, and if one really wants to see dry animal tissue, tissues from birds of prey (hawks), other birds and reptiles can be among the worst. We custom process our rodent tissues, brain, small, etc and have shorter schedules for mouse brain versus hamster brain, that type of thing just to avoid overdehydration. The 1 hour per station or change even with 3 X 95% and 3 x 100% never seems to bother human species tissues much, but with rodent tissue, it removes too much of the bound water. some larger animals with bigger samples in a cassette seem to be ok with standard processing schedules but never with heat added. A curious thing observed over a long career in histowork, I saw the addition of heat to processing in automated processor a curious thing. When we used the older carousel (Sp?) models i.e Technicon, heat couldn't be added and processing was done at RT. True, heat may speed up the solvent exchange but it also can add to drying of tissues. I have always wondered why this has become a standard method over the years other than trying to speed up processing, but always felt alternating vacuum and pressure to be the best way to do that along with paying attention to length of processing schedules. What one needs to do is find the correct balance so that the free water in the tissue spaces is removed and not the bound water on protein molecules. Overdehydration and adding heat to processing will exacerbate the removal of bound water. Consequently, heat is never added to our animal tissue processing steps. At 02:38 AM 2/1/2006, you wrote: >Think rodent tissue can process on the 'hard' side. Wonder what makes it >'hard'? Is it that the bonds between the proteins are 'stronger' or the >proteins are 'nearer' together? I mean if we could figure out what made >tissue 'hard' molecularly then we'd know which the culprit in the processing >was. > >Sorry to ramble. > >Kemlo Rogerson >Pathology Manager >Ext 3311 >DD 01934 647057 >Mob 07749 754194 > > > > >-----Original Message----- >From: Steven Coakley [mailto:sjchtascp@yahoo.com] >Sent: Tuesday, January 31, 2006 6:56 PM >To: Histonet@lists.utsouthwestern.edu >Subject: [Histonet] processing > >Good afternoon, > > I'm working with mouse/rat muscle and trying to fine tune my processing >schecdule. > My last run of muscle were very dry. I have a copy of the NSH Animal >Processing Manual and trying to strike a balance between those listed on >page 5-6. I've noticed most of the protocols do not call for heat or P/V. >The scedule I currently use is as follows: 1-70% (hold), 1-80%, 2-95%, >3-100%, 3-xylene all for 30 minutes each and at 38C with P/V. 3 paraffins, >45 minutes each at 55C with P/V. I'm fairly new to research and still >trying to make the "mental" switch in handling the different tissue. > > Thanks everyone, > > Steve > > > >--------------------------------- >Bring words and photos together (easily) with > PhotoMail - it's free and works with Yahoo! Mail. >_______________________________________________ >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 Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 19 Date: Thu, 2 Feb 2006 05:47:20 -0600 From: "Molinari, Betsy" Subject: RE: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections To: Message-ID: Content-Type: text/plain; charset="us-ascii" Once a month I put a calibrated thermometer in one of the paraffin reservoirs. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Porter Sent: Wednesday, February 01, 2006 3:13 PM To: Histonet Subject: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections If your willing to share - how are people monitoring the temperature of paraffin baths on tissue processors? Are you using additional thermometers directly into the paraffin baths, or just going by the digital read out on the display for the (oven on a VIP) instrument? Just curious how everyone is dealing with this particular item. Thanks in advance for the many responses and ideas I know I will get from asking this question! Amy S. Porter, HT(ASCP) QIHC Laboratory Supervisor Michigan State University Investigative HistoPathology Laboratory Department of Physiology / Division of Human Pathology 2100 Biomedical Physical Sciences Bldg. Room #2133 East Lansing, MI 48824-3320 Phone: (517) 355-6475 ext 1480 / Fax: (517) 432-1368 Email: portera@msu.edu www.humanpathology.msu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 20 Date: Thu, 2 Feb 2006 13:29:53 +0000 From: Susan.Ferrigon@sanofi-aventis.com Subject: [Histonet] Kuppfer cells To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Hi Can anyone suggest a marker for Kuppfer cells?? Thanks Susan ------------------------------ Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de ce courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuellement pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ------------------------------ ------------------------------ Message: 21 Date: Thu, 2 Feb 2006 09:43:58 -0500 From: "Adams, Nancy" Subject: [Histonet] microwave transparent containers and equipment To: Message-ID: <17A1862099540D458C8FE9380C2BC461C0932E@fh2xmail.fhdomain1.capecodhealth.org > Content-Type: text/plain Good morning Does the new CAP question ANP.28860 require documentation that the containers and equipment are, indeed, microwave transparent? Somehow I think just saying yes isn't going to be enough:-) Thanks for your thoughts. Nancy Rutledge Falmouth Hospital ************************************************************ This email and any files transmitted with it are confidential. And intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please contact the system administrator for Cape Cod Healthcare. Administrator@CapeCodHealth.org ************************************************************ ------------------------------ Message: 22 Date: Thu, 2 Feb 2006 07:28:05 -0800 (PST) From: GT Hebert Subject: Re: [Histonet] Kuppfer cells To: Susan.Ferrigon@sanofi-aventis.com Cc: histonet@lists.utsouthwestern.edu Message-ID: <20060202152805.96087.qmail@web31704.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Susan, I've used F4/80. Though I was trying to stain macrophages, I used the Kupffer cells in the liver as a positive control to show that the antibody was working and they stained perfectly. Gustave Hebert Scientist II Wyeth Research Cambridge MA Susan.Ferrigon@sanofi-aventis.com wrote: Hi Can anyone suggest a marker for Kuppfer cells?? Thanks Susan ------------------------------ Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de ce courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuellement pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Autos. Looking for a sweet ride? Get pricing, reviews, & more on new and used cars. ------------------------------ Message: 23 Date: Thu, 2 Feb 2006 07:44:20 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] microwave transparent containers and equipment To: "Adams, Nancy" , histonet@lists.utsouthwestern.edu Message-ID: <20060202154420.27452.qmail@web61225.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Nancy: You could show them a list with the microwaves penetration (cm) of different substances. For example: polyethyl methacrylate = 300 cm (= 3 m) Teflon = 9,000 cm (= 90 m) Paraffin wax = 15,000 (=150 m) Styrofoam= 40,000 cm (=400 m) Showing them the materials your containers are made of I think that they will accept that they are indeed transparent. Hope this will help. Ren? J. "Adams, Nancy" wrote: Good morning Does the new CAP question ANP.28860 require documentation that the containers and equipment are, indeed, microwave transparent? Somehow I think just saying yes isn't going to be enough:-) Thanks for your thoughts. Nancy Rutledge Falmouth Hospital ************************************************************ This email and any files transmitted with it are confidential. And intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please contact the system administrator for Cape Cod Healthcare. Administrator@CapeCodHealth.org ************************************************************ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? With a free 1 GB, there's more in store with Yahoo! Mail. ------------------------------ Message: 24 Date: Thu, 2 Feb 2006 17:09:19 +0100 From: "Bruijntjes, J.P." Subject: FW: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections To: Message-ID: <3B070848E7C2204F9DEB8BCFD767728004A69837@ntexch1.voeding.tno.nl> Content-Type: text/plain; charset="us-ascii" Hi Amy We use an electronic thermometer (testo 915-1) which is calibrated once or twice a year. Joost Bruijntjes TNO Quality of Life Toxicology and Applied Pharmacology Zeist Holland -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Molinari, Betsy Sent: donderdag 2 februari 2006 12:47 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections Once a month I put a calibrated thermometer in one of the paraffin reservoirs. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Porter Sent: Wednesday, February 01, 2006 3:13 PM To: Histonet Subject: [Histonet] Tissue Processor Paraffin Temps for CAP Inspections If your willing to share - how are people monitoring the temperature of paraffin baths on tissue processors? Are you using additional thermometers directly into the paraffin baths, or just going by the digital read out on the display for the (oven on a VIP) instrument? Just curious how everyone is dealing with this particular item. Thanks in advance for the many responses and ideas I know I will get from asking this question! Amy S. Porter, HT(ASCP) QIHC Laboratory Supervisor Michigan State University Investigative HistoPathology Laboratory Department of Physiology / Division of Human Pathology 2100 Biomedical Physical Sciences Bldg. Room #2133 East Lansing, MI 48824-3320 Phone: (517) 355-6475 ext 1480 / Fax: (517) 432-1368 Email: portera@msu.edu www.humanpathology.msu.edu _______________________________________________ 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 This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html ------------------------------ Message: 25 Date: Thu, 02 Feb 2006 10:02:18 -0700 From: Gayle Callis Subject: Re: [Histonet] citation for negative controls To: "Jonathan Wilson" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20060202095040.01b3ce58@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Appropriate negative controls are discussed by Boenisch, in DAKO 3rd Edition Handbook Immunochemical Staining methods, and found on the DAKO website as a free pdf download. It can be done and clinical laboratories do use irrelevant antibodies at times also. Jules Elias books also discuss this, Immunohistopathology, a pracatical approach to diagnostics, ASCP press. There is a second edition although may not be available to you. If you are working with mouse, this can be a difficult thing to find at times so that you get NO staining with an irrelevant antibody. Consequently we use what is most cited in the literature, isotype matched immunoglobulin i.e. IgG controls depending on species and immunoglobulin. They come pure, biotinylated, conjugated to many things including Alexa dyes. OR you can buy whole IgG's from Jackson (whole rat IgG contains all the isotypes, and from many different species.) One that happened some time ago, we used rat serum as a control and the reviewers balked big time. We had to REPEAT what was a very difficult, tedious immunogold staining project using isotype matched controls. Since that time, we NEVER use normal serums nor an irrelevant antibody for a negative control. Having to repeat work is not fun! At 04:34 PM 2/1/2006, you wrote: >Hello, >I use IHC in basic research and have been digging through the archives for >information on appropriate negative controls and came across the use of >irrelavent antibodies. I would like to use this information in a paper I >am writing and was wondering if this is cited in a review or paper. I have >Harlow and Lane's book but it doesn't mention this type of control. >Thank you in advance for any help. >Sincerely, >Jon > > >Jonathan Wilson (PhD) >ecofisiologia CIMAR >Rua dos Bragas 289 >4050-123 Porto Portugal >office 351 22 340 1809 >lab 351 22 340 1834 >fax 351 22 339 0608 >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 27, Issue 2 *************************************** From Susan.Ferrigon <@t> sanofi-aventis.com Fri Feb 3 06:18:29 2006 From: Susan.Ferrigon <@t> sanofi-aventis.com (Susan.Ferrigon@sanofi-aventis.com) Date: Fri Feb 3 06:18:42 2006 Subject: [Histonet] Kuppfer cells Message-ID: Hi Does anyone know of anF4/80 marcrophage marker that will work on rat tissue. Thanks Susan |---------+---------------------------> | | GT Hebert | | | | | | | | | 02/02/2006 15:28| | | | |---------+---------------------------> >-------------------------------------------------------------------------------------------------------------------------------| | | | To: Susan Ferrigon/GB-ALNWICK/RESEARCH/SANOFI@Research | | cc: histonet@lists.utsouthwestern.edu | | Subject: Re: [Histonet] Kuppfer cells | >-------------------------------------------------------------------------------------------------------------------------------| Susan, I've used F4/80. Though I was trying to stain macrophages, I used the Kupffer cells in the liver as a positive control to show that the antibody was working and they stained perfectly. Gustave Hebert Scientist II Wyeth Research Cambridge MA Susan.Ferrigon@sanofi-aventis.com wrote: Hi Can anyone suggest a marker for Kuppfer cells?? Thanks Susan ------------------------------ Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de ce courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuellement pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Yahoo! Autos. Looking for a sweet ride? Get pricing, reviews, & more on new and used cars. ------------------------------ Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de ce courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuellement pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ------------------------------ From petepath <@t> yahoo.com Fri Feb 3 07:02:01 2006 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Feb 3 07:02:10 2006 Subject: [Histonet] What are peoples feelings on pre labling slides? Message-ID: <20060203130201.24018.qmail@web30408.mail.mud.yahoo.com> I am curious to see if it is considered acceptable practice to pre-label multiple slides before cutting the blocks and picking the tissues up on these pre-labeled slides. We came close to a dangerous misdiagnosis because a tech picked up a malignant section from a " part 2 breast biopsy" on a prelabled part 1 slide. Luckily it made no sense that only one of many slides contained tumor that looked like it was coming from an advanced tumor. After playing match the blocks it was obvious that the malignant part one slide matched a part 2 block. It seems to me that this is a potentially dangerous habit despite the convenience of assembly line labeling. Early in my career I stopped labeling my frozen section slides up front and wait until after I pick up the section. When I am cutting frozens I make variable #s of slides depending on the situation. Working quickly under the pressure of multiple cases it is not hard to pick up the wrong slide and make this mistake. I am curious to hear peoples thoughts. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From jqb7 <@t> cdc.gov Fri Feb 3 07:15:44 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Feb 3 07:14:24 2006 Subject: [Histonet] Leica stainer/coverslipper combo. Message-ID: Hello all: Has anyone tried the Leica Autostainer XL ST5010 Workstation? What do you like/dislike about the unit? Thanks, Jeanine Bartlett, BS, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov From GauchV <@t> mail.amc.edu Fri Feb 3 07:18:26 2006 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Fri Feb 3 07:18:49 2006 Subject: [Histonet] What are peoples feelings on pre labling slides? Message-ID: We do not pre-label any of our slides in the lab. We cut our slides and then label them as we do each one. As for frozens, we label the slide as we cut the frozen- if we do multiple frozens on that block we label each one as we go. It has been our policy to do it this way for a long time now..in the past we had a few techs who decided to do it "their" way instead of following our procedures and it almost resulted in a case being sent out mislabeled (fortunately it ws caught before it left the lab) which is why we do not allow that practice in our lab. Vicki Gauch AMCH Albany, NY >>> "Stephen Peters M.D." 2/3/2006 8:02:01 AM >>> I am curious to see if it is considered acceptable practice to pre-label multiple slides before cutting the blocks and picking the tissues up on these pre-labeled slides. We came close to a dangerous misdiagnosis because a tech picked up a malignant section from a " part 2 breast biopsy" on a prelabled part 1 slide. Luckily it made no sense that only one of many slides contained tumor that looked like it was coming from an advanced tumor. After playing match the blocks it was obvious that the malignant part one slide matched a part 2 block. It seems to me that this is a potentially dangerous habit despite the convenience of assembly line labeling. Early in my career I stopped labeling my frozen section slides up front and wait until after I pick up the section. When I am cutting frozens I make variable #s of slides depending on the situation. Working quickly under the pressure of multiple cases it is not hard to pick up the wrong slide and make this mistake. I am curious to hear peoples thoughts. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------- CONFIDENTIALITY NOTICE: This email and any attachments may contain confidential information that is protected by law and is for the sole use of the individuals or entities to which it is addressed. If you are not the intended recipient, please notify the sender by replying to this email and destroying all copies of the communication and attachments. Further use, disclosure, copying, distribution of, or reliance upon the contents of this email and attachments is strictly prohibited. To contact Albany Medical Center, or for a copy of our privacy practices, please visit us on the Internet at www.amc.edu. From jqb7 <@t> cdc.gov Fri Feb 3 07:19:58 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Feb 3 07:19:16 2006 Subject: [Histonet] RE: Leica stainer/coverslipper combo. Message-ID: I may have that model number wrong: it might be TS 5025. At any rate, it combines the St 5020 stainer and the CV 5030 coverslipper. Jeanine Bartlett, BS, HT(ASCP) (404) 639-3590 jeanine.bartlett@cdc.hhs.gov > _____________________________________________ > From: Bartlett, Jeanine > Sent: Friday, February 03, 2006 8:16 AM > To: Histonet (histonet@lists.utsouthwestern.edu) > Subject: Leica stainer/coverslipper combo. > Sensitivity: Confidential > > Hello all: > > Has anyone tried the Leica Autostainer XL ST5010 Workstation? What do > you like/dislike about the unit? > > Thanks, > > Jeanine Bartlett, BS, HT(ASCP) > Centers for Disease Control and Prevention > 1600 Clifton Road, MS/G-32 > 18/SB-114 > Atlanta, GA 30333 > (404) 639-3590 > jeanine.bartlett@cdc.hhs.gov > From rjbuesa <@t> yahoo.com Fri Feb 3 07:22:41 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Feb 3 07:22:50 2006 Subject: [Histonet] What are peoples feelings on pre labling slides? In-Reply-To: <20060203130201.24018.qmail@web30408.mail.mud.yahoo.com> Message-ID: <20060203132241.62804.qmail@web61215.mail.yahoo.com> Stephen: My personal experience: In the labs I have supervised all those histotechs that used to label their own slides, did the labelling after the blocks were trimmed (faced off) and were cooling in the tray. To allow a better cooling they used to hand write the labels that they organized according with the blocks that were cut immediately after. There were histotechs that used to label the slide after they picked up the sections from the water bath while the block was still in the microtome. Other large laboratory I supervised had slides writing machines and the slides were written before the tissues were even finished processing. A lab aide did the writing and another matched slides with blocks that were handed out to the histotechs. I know of supervisors that say it is a no-no to prelabel slides. In my experience any mismatch is due to the histotech and not to the system. In the same way that any dislexic histotech can write the wrong number before or after the section has been cut. Everything boils down to the care and attention the histotecs pays to his/her work, not when the slides are written. Hope this will help. Ren? J. "Stephen Peters M.D." wrote: I am curious to see if it is considered acceptable practice to pre-label multiple slides before cutting the blocks and picking the tissues up on these pre-labeled slides. We came close to a dangerous misdiagnosis because a tech picked up a malignant section from a " part 2 breast biopsy" on a prelabled part 1 slide. Luckily it made no sense that only one of many slides contained tumor that looked like it was coming from an advanced tumor. After playing match the blocks it was obvious that the malignant part one slide matched a part 2 block. It seems to me that this is a potentially dangerous habit despite the convenience of assembly line labeling. Early in my career I stopped labeling my frozen section slides up front and wait until after I pick up the section. When I am cutting frozens I make variable #s of slides depending on the situation. Working quickly under the pressure of multiple cases it is not hard to pick up the wrong slide and make this mistake. I am curious to hear peoples thoughts. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Bring words and photos together (easily) with PhotoMail - it's free and works with your Yahoo! Mail. From oshel1pe <@t> cmich.edu Fri Feb 3 07:27:33 2006 From: oshel1pe <@t> cmich.edu (Oshel, Philip Eugene) Date: Fri Feb 3 07:27:53 2006 Subject: [Histonet] ?s4 electron microscopists Message-ID: <07BD45BB3EE8A0468C94776E7A0C660E3A335D@cmail4.central.cmich.local> Rita, There are pros and cons. Film is still higher resolution than digital, with more grey levels, but digital is high enough resolution to produce publishable prints, and getting better. Other choices are to take film images and then digitize them on a flatbed scanner, or to use imaging plates. Imaging plates produces digital images using phosphor-coated plates. The images are equal-equal to film, or are as good as film, but require special scanners, etc. A digital camera is the easiest, and it is faster and cheaper to deal with digital images than to deal with film. Also, there is no expense for darkrooms, darkroom chemicals, and hazardous waste. Digital images are typically 1 to 4MB (can be bigger), depending on the number of pixels in the acquired image, the bit depth (8-bit or 16-bit for example) and the imaging program. 1024 X 1024 is 1 to 2MB, e.g. Bit depth is usually 8, 12, or 16, but most imaging programs are 8-bit. Some can do 16-bit. The main factor with the imaging program is to be sure it will save images *with* the relevant information (kV, mag, etc.) imprinted on the image *as a TIFF file*, not only as some proprietary format that can only be read by the imaging program. The image must be readable by e.g., Irfanview (shareware for PCs free for non-profit use, rudimentary image processing), Graphic Converter (for Macs, shareware, almost as powerful as Photoshop and easier to use), ImageJ, or Photoshop. Irfanview and Graphic Converter are also file format conversion programs. A major issue is that digital imaging has no negative. This means that the original image can be manipulated and lost. Even such an innocent thing as changing the brightness and contrast usually changes the actual data. Recall that a digital image is a N x M array of pixels (e.g. 1024 x 1024), each pixed occupied by a number, which represents brightness (intensity). So altering even such basic settings as brightness changes the number, therefore the image. Altering "Levels" changes the *display* of the image, so is a better thing to do. But what *must* be drilled into people's heads is to burn the data onto a NONrewritable CD *before* doing *anything* to the image. This then becomes the photographic negative, and if a copy is manipulated in any way, the original image is still available unaltered. People *will* manipulate digital images. It's very easy to do, and doesn't require much if any understanding how the manipulations are done, or what they do to the original images. So greater precautions must be taken to save the original images in an unalterable form. Emailing files is simple and no big deal. Mind, your Clinic IT people can make it a horrible headache, but I routinely email digital micrographs around the country. You could also set up a computer as a ftp server on a local network, put the images on that, and let the pathologists download the images from it. Not as convenient as email, but neither will they get lost in emailboxes that they never empty. Plus, the original image is left on the ftp server, and can then be burned to a CD. (Just spend the extra penny to get good CDs! Cheap ones last a few years, good ones are archival.) Digital cameras for TEMs are generally very expensive, but mostly because the companies have been getting away with the high prices. I'd suggest contacting SIA, Scientific Instruments and Applications: http://www.sia-cam.com/ (I have no business interest in SIA, nor am I a customer, but if I get to digitize our Philips CM-10, SIA is where I'm likely to go. Their prices are much more reasonable for the same quality cameras, and their service reputation is better.) Phil Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University Mt. Pleasant, MI 48859 (989) 774-3576 Subject: [Histonet] ?s4 electron microscopists Dear histonet electron microscopists - I am currently being trained in to EM to take over for our retiring microscopist. We are building a new lab, and the administration's plan is to go digital. (I have a Zeiss 900 TEM) Are any of you out there using a digital camera on your TEM? What are the advantages and disadvantages? Is the clarity comparable to a darkroom print? The theory is that I will upload my images to my computer, then email them to the pathologists ~ with the huge amount of pixels required, won't this take an awfully long time for the pathologists to download the images? Any thoughts will be greatly appreciated. Thanks! Rita Kenner HTL (ASCP) Marshfield Clinic Marshfield, WI _____________________________________________________________ Netscape. Just the Net You Need. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Diane.Gladney <@t> se.amedd.army.mil Fri Feb 3 07:45:22 2006 From: Diane.Gladney <@t> se.amedd.army.mil (Gladney, Diane C Ms MACH) Date: Fri Feb 3 07:41:52 2006 Subject: [Histonet] What are peoples feelings on pre labling slides? Message-ID: <4D55B2E997EFAE4DA6081DDE100B8302B78181@amedmlsermc133.amed.ds.army.mil> I agree with Rene. It is about attention to detail. I work in a small lab and we pre-label our slides. I confirm the case and block number with my prelabeled slides before I cut the block. This helps to eliminate picking up the wrong slide. If a tech is rushed to get the work out (whether using pre-labeled slides or not) and does not pay attention to the detail of the surgical number and block number then it is on the tech and not on the system used. Thanks, Diane Diane C. Gladney, HT (ASCP) Supervisor, Histology/Cytology Moncrief Army Community Hospital Dept. of Pathology 4500 Stuart Street P.O. Box 484 Ft. Jackson, SC 29207 Email: diane.gladney@se.amedd.army.mil Phone: 803-751-2530 FAX: 803-751-7829 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Friday, February 03, 2006 8:23 AM To: Stephen Peters M.D.; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] What are peoples feelings on pre labling slides? Stephen: My personal experience: In the labs I have supervised all those histotechs that used to label their own slides, did the labelling after the blocks were trimmed (faced off) and were cooling in the tray. To allow a better cooling they used to hand write the labels that they organized according with the blocks that were cut immediately after. There were histotechs that used to label the slide after they picked up the sections from the water bath while the block was still in the microtome. Other large laboratory I supervised had slides writing machines and the slides were written before the tissues were even finished processing. A lab aide did the writing and another matched slides with blocks that were handed out to the histotechs. I know of supervisors that say it is a no-no to prelabel slides. In my experience any mismatch is due to the histotech and not to the system. In the same way that any dislexic histotech can write the wrong number before or after the section has been cut. Everything boils down to the care and attention the histotecs pays to his/her work, not when the slides are written. Hope this will help. Ren? J. "Stephen Peters M.D." wrote: I am curious to see if it is considered acceptable practice to pre-label multiple slides before cutting the blocks and picking the tissues up on these pre-labeled slides. We came close to a dangerous misdiagnosis because a tech picked up a malignant section from a " part 2 breast biopsy" on a prelabled part 1 slide. Luckily it made no sense that only one of many slides contained tumor that looked like it was coming from an advanced tumor. After playing match the blocks it was obvious that the malignant part one slide matched a part 2 block. It seems to me that this is a potentially dangerous habit despite the convenience of assembly line labeling. Early in my career I stopped labeling my frozen section slides up front and wait until after I pick up the section. When I am cutting frozens I make variable #s of slides depending on the situation. Working quickly under the pressure of multiple cases it is not hard to pick up the wrong slide and make this mistake. I am curious to hear peoples thoughts. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Bring words and photos together (easily) with PhotoMail - it's free and works with your Yahoo! Mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo.rogerson <@t> waht.swest.nhs.uk Fri Feb 3 08:01:37 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Fri Feb 3 08:01:52 2006 Subject: [Histonet] What are peoples feelings on pre labling slides? Message-ID: Pre-labelling of slides both for Histo and Cyto, pre-labelling of blood bottles and LBC vials, in my opinion wastes more time than it saves because of the cock ups and litigation it creates. My old Senior Chief used to say you should treat every specimen, slide, etc. as if it were from you or one of your relatives. Obviously if you hate your relatives or have a death wish then the example doesn't work. Personally, I wouldn't, pre-label slides that is. Kemlo Rogerson Weston General Hospital Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Stephen Peters M.D. [mailto:petepath@yahoo.com] Sent: Friday, February 03, 2006 1:02 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] What are peoples feelings on pre labling slides? I am curious to see if it is considered acceptable practice to pre-label multiple slides before cutting the blocks and picking the tissues up on these pre-labeled slides. We came close to a dangerous misdiagnosis because a tech picked up a malignant section from a " part 2 breast biopsy" on a prelabled part 1 slide. Luckily it made no sense that only one of many slides contained tumor that looked like it was coming from an advanced tumor. After playing match the blocks it was obvious that the malignant part one slide matched a part 2 block. It seems to me that this is a potentially dangerous habit despite the convenience of assembly line labeling. Early in my career I stopped labeling my frozen section slides up front and wait until after I pick up the section. When I am cutting frozens I make variable #s of slides depending on the situation. Working quickly under the pressure of multiple cases it is not hard to pick up the wrong slide and make this mistake. I am curious to hear peoples thoughts. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bob-meyer <@t> northwestern.edu Fri Feb 3 08:09:20 2006 From: bob-meyer <@t> northwestern.edu (bob-meyer@northwestern.edu) Date: Fri Feb 3 08:09:28 2006 Subject: [Histonet] Alkaline Phosphatase Message-ID: <20060203140920.6B9A735C4C@casbah.it.northwestern.edu> Lorie, First thing is to check the temperature of the oven with a thermometer to make sure it is truly at 60 C. It is true that high heat in the oven can destroy antigenicity although the debate is still out there of why doesn't the high heat of antigen retrieval destroy antigenicity but rather improves it. Anyways, another method of adhering sections to the slide without the use of heat would be to use silanized slides (we use the ones from Dako) and allow to air dry over night at room temperature. Bob Meyer Northwestern University ==============Original message text=============== On Thu, 02 Feb 2006 10:31:45 pm +0000 bjdewe@aol.com wrote: Hi all, Maybe you can give me some advice. I have been staining some bone tissue for alkaline phosphatase using a Bone Alk Phos antibody. I was getting reasonable staining until recently after I started putting the tissue in a 60degree C oven to help adhere the tissue to the + slides. I was having some trouble with the sections coming off. I'm now wondering if my problem is the heat on the tissue making the AlkPhos inactive. Comments? TIA... Lorie ******************************************************************* Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s)and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ******************************************************************* _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ===========End of original message text=========== From Jacqueline.Malam <@t> rli.mbht.nhs.uk Fri Feb 3 08:38:59 2006 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Feb 3 08:43:31 2006 Subject: [Histonet] Negative controls Message-ID: Dako have a good section about controls in their handbook and it's also in Immunocytochemistry by Julia M Polak. Jacqui Malam Lancaster UK DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From RBARNHART <@t> summithealth.org Fri Feb 3 08:52:50 2006 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Feb 3 08:53:26 2006 Subject: [Histonet] What are peoples feelings on pre labeling slides? Message-ID: I agree with Diane and Rene. We pre label our slides and I personally have not had problems with picking up the wrong tissue on the slide because I check the case number and block number before I pick up the slide. After all that is my responsibility to make sure what is on the slide, pre labeled or not, is the correct tissue. We use to hand write the case number on the slide then put a label on after staining. We had a tech that on different occasions put the wrong label on because she was not paying attention to the case number that was hand written. My feeling is we can not be on autopilot and if a tech is not paying attention when they are picking up a section on a pre labeled slide then there is a chance they will mislabel the slide anyway. Just my opinion. Becky >>> "Gladney, Diane C Ms MACH" 2/3/2006 8:45:22 AM >>> I agree with Rene. It is about attention to detail. I work in a small lab and we pre-label our slides. I confirm the case and block number with my prelabeled slides before I cut the block. This helps to eliminate picking up the wrong slide. If a tech is rushed to get the work out (whether using pre-labeled slides or not) and does not pay attention to the detail of the surgical number and block number then it is on the tech and not on the system used. Thanks, Diane Diane C. Gladney, HT (ASCP) Supervisor, Histology/Cytology Moncrief Army Community Hospital Dept. of Pathology 4500 Stuart Street P.O. Box 484 Ft. Jackson, SC 29207 Email: diane.gladney@se.amedd.army.mil Phone: 803-751-2530 FAX: 803-751-7829 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Friday, February 03, 2006 8:23 AM To: Stephen Peters M.D.; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] What are peoples feelings on pre labling slides? Stephen: My personal experience: In the labs I have supervised all those histotechs that used to label their own slides, did the labelling after the blocks were trimmed (faced off) and were cooling in the tray. To allow a better cooling they used to hand write the labels that they organized according with the blocks that were cut immediately after. There were histotechs that used to label the slide after they picked up the sections from the water bath while the block was still in the microtome. Other large laboratory I supervised had slides writing machines and the slides were written before the tissues were even finished processing. A lab aide did the writing and another matched slides with blocks that were handed out to the histotechs. I know of supervisors that say it is a no-no to prelabel slides. In my experience any mismatch is due to the histotech and not to the system. In the same way that any dislexic histotech can write the wrong number before or after the section has been cut. Everything boils down to the care and attention the histotecs pays to his/her work, not when the slides are written. Hope this will help. Ren? J. "Stephen Peters M.D." wrote: I am curious to see if it is considered acceptable practice to pre-label multiple slides before cutting the blocks and picking the tissues up on these pre-labeled slides. We came close to a dangerous misdiagnosis because a tech picked up a malignant section from a " part 2 breast biopsy" on a prelabled part 1 slide. Luckily it made no sense that only one of many slides contained tumor that looked like it was coming from an advanced tumor. After playing match the blocks it was obvious that the malignant part one slide matched a part 2 block. It seems to me that this is a potentially dangerous habit despite the convenience of assembly line labeling. Early in my career I stopped labeling my frozen section slides up front and wait until after I pick up the section. When I am cutting frozens I make variable #s of slides depending on the situation. Working quickly under the pressure of multiple cases it is not hard to pick up the wrong slide and make this mistake. I am curious to hear peoples thoughts. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Bring words and photos together (easily) with PhotoMail - it's free and works with your Yahoo! Mail. _______________________________________________ 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 From gu.lang <@t> gmx.at Fri Feb 3 08:59:48 2006 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Feb 3 08:59:52 2006 Subject: AW: [Histonet] What are peoples feelings on pre labling slides? In-Reply-To: <20060203130201.24018.qmail@web30408.mail.mud.yahoo.com> Message-ID: <000901c628d2$7aa71010$eeeea8c0@SERVER01> In our lab we do prelabbeling for the next 10 blocks to cut. We have them in a box and take one after the other. Labelling after cutting is the wrong direction in my feeling/opinion. Handling a slide with the unprotected section on it had to be very carefully. Prelabelling is faster and doesn't cause more mistakes than "after-labelling". The slide is dry and easier to label. With the label-systems (barcodes) for eg. Immunhisto it is laborious to do a "double-labelling". The point is the resposibility an accuracy of the tech before or/and after cutting. Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Stephen Peters M.D. Gesendet: Freitag, 03. Februar 2006 14:02 An: Histonet@lists.utsouthwestern.edu Betreff: [Histonet] What are peoples feelings on pre labling slides? I am curious to see if it is considered acceptable practice to pre-label multiple slides before cutting the blocks and picking the tissues up on these pre-labeled slides. We came close to a dangerous misdiagnosis because a tech picked up a malignant section from a " part 2 breast biopsy" on a prelabled part 1 slide. Luckily it made no sense that only one of many slides contained tumor that looked like it was coming from an advanced tumor. After playing match the blocks it was obvious that the malignant part one slide matched a part 2 block. It seems to me that this is a potentially dangerous habit despite the convenience of assembly line labeling. Early in my career I stopped labeling my frozen section slides up front and wait until after I pick up the section. When I am cutting frozens I make variable #s of slides depending on the situation. Working quickly under the pressure of multiple cases it is not hard to pick up the wrong slide and make this mistake. I am curious to hear peoples thoughts. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Feb 3 09:17:21 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Feb 3 09:17:42 2006 Subject: [Histonet] paraffin embedding of frozen tissue In-Reply-To: References: Message-ID: <6.0.0.22.1.20060203080621.01b54610@gemini.msu.montana.edu> When working with mouse lung, it is best to fill the lung with OCT, then snap freeze. OR fill the lung with 2.5 ml fixative, then cryopreserve with 30% sucrose after fixation is finished, snap freeze. We get perfect lung sections when we do this. Filling is done - anesthetize the mouse or you can euthanize but no by cervical spinal cord dislocation. Open chest and up under chin, expose trachea very carefully. Make a tiny V shaped cut in TOP of trachea, then you can introduce an 19 guage dulled hypodermic needle attached to a syringe filled with either OCT or if you prefer fixative. Inject only 2.5 ml (use a 3 ml syringe for this) if and watch the lung inflate. AVOID overinflation of lung, it will blow alveoli away. The OCT is NOT diluted, and whatever you do, do NOT severe the trachea or you cannot fill the lung - the trachea will retract into chest cavitity and you will fish around for it. We use very sharp pointed curved cuticle scissors found in drug stores - these are stainless steel with the finest points and make excellent cut on top of trachea. We clamp off the trachea where needle goes in using a mosquito hemostat, and carefully dissect the lung out, lifting carefully by trachea, remove heart, and snap freeze embedded in OCT (you can remove any lobe you wish at this point by laying filled lung on back of a Petri dish). Some people perfuse the lung with fixative via heart, but you must severe the descending arteries behind the intestines - 18 guage or smaller needle with 10 ml syringe filled with fixative works. At 02:33 PM 2/2/2006, you wrote: >Greetings histonetters, >I have recently found that sectioning frozen lung is a bit difficult. I >was wondering if anyone has ever paraffin embedded previously fixed (and >cryproserved) frozen sections? And if so what were the results? I am >performing immunohistochemistry / immunofluorecent staining on mice lung >tissue and I was just wondering if it was possible. I also encourge >advice on the subject matter. > > >I have learned quite a bit from this web site. >Thank you very much. >Carmen Contreras-Sesvold > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From ROrr <@t> enh.org Fri Feb 3 09:23:54 2006 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Fri Feb 3 09:24:05 2006 Subject: [Histonet] MSH6 Message-ID: Hi Josie, How many different types of protocols did you try on the XT? With the different temp and Cell conditioning options I'm surprised you didn't get something. Are you running the MLH-1 and MSH-2 as well? How are they? My experience with the Microsateliite Instabilities is 4+ staining in 2 out of 3 antibodies, depending on the control tissue. The third antibody would compare at 2+ or a bit stronger. I have the best of both worlds with a Benchmark and a Nemesis. I experienced the poor staining results of these particular antibodies and I decided to run them on the Nemesis with Biocare polymer detection and they are gorgeous. I use the Biocare antibodies, too. In case you don't have that option, I would consider trying another control tissue, and running through all the CC options. I'm not sure if they behave differently without heat, did you try running a protocol with out heat in the detection? Do you have access to the polymer from Ventana? Maybe try that? Or run the antibody at 1:5 or 1:10. Or do you have a pressure cooker to try the HEIR offline. If you get good staining with offline HEIR then you can rule out the detection as the issue, too. Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 -----Original Message----- Message: 8 Date: Wed, 1 Feb 2006 13:54:36 -0600 From: "Nava, Josefa" Subject: [Histonet] looking for Antibodies-MSH6 and PMS2 To: Message-ID: <2C515C1049EAF5459EFD8C9B929078A41945A8@phdex03.txhealth.org> Content-Type: text/plain; charset="us-ascii" Hello Everyone , I am using Ventana BMK/XT , can someone tell me a good source of MSH6 and PMS2 that will work on the Ventana Machine. I appreciate any information. Thank you. Josie From bwhitaker <@t> brownpathology.com Fri Feb 3 09:27:14 2006 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Feb 3 09:27:19 2006 Subject: [Histonet] What are peoples feelings on pre lablingslides? In-Reply-To: Message-ID: <001501c628d6$4f799bc0$3601a8c0@brownpathology.net> IMHO, ideally, someone is matching blocks and slides before they go out (part of the QA process). I know that it doesn't ALWAYS happen, but I think that regardless of when the slides are labeled, checking them is a prudent thing to do. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vicki Gauch Sent: Friday, February 03, 2006 7:18 AM To: Histonet@lists.utsouthwestern.edu; petepath@yahoo.com Subject: Re: [Histonet] What are peoples feelings on pre lablingslides? We do not pre-label any of our slides in the lab. We cut our slides and then label them as we do each one. As for frozens, we label the slide as we cut the frozen- if we do multiple frozens on that block we label each one as we go. It has been our policy to do it this way for a long time now..in the past we had a few techs who decided to do it "their" way instead of following our procedures and it almost resulted in a case being sent out mislabeled (fortunately it ws caught before it left the lab) which is why we do not allow that practice in our lab. Vicki Gauch AMCH Albany, NY >>> "Stephen Peters M.D." 2/3/2006 8:02:01 AM >>> I am curious to see if it is considered acceptable practice to pre-label multiple slides before cutting the blocks and picking the tissues up on these pre-labeled slides. We came close to a dangerous misdiagnosis because a tech picked up a malignant section from a " part 2 breast biopsy" on a prelabled part 1 slide. Luckily it made no sense that only one of many slides contained tumor that looked like it was coming from an advanced tumor. After playing match the blocks it was obvious that the malignant part one slide matched a part 2 block. It seems to me that this is a potentially dangerous habit despite the convenience of assembly line labeling. Early in my career I stopped labeling my frozen section slides up front and wait until after I pick up the section. When I am cutting frozens I make variable #s of slides depending on the situation. Working quickly under the pressure of multiple cases it is not hard to pick up the wrong slide and make this mistake. I am curious to hear peoples thoughts. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------- CONFIDENTIALITY NOTICE: This email and any attachments may contain confidential information that is protected by law and is for the sole use of the individuals or entities to which it is addressed. If you are not the intended recipient, please notify the sender by replying to this email and destroying all copies of the communication and attachments. Further use, disclosure, copying, distribution of, or reliance upon the contents of this email and attachments is strictly prohibited. To contact Albany Medical Center, or for a copy of our privacy practices, please visit us on the Internet at www.amc.edu. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo.rogerson <@t> waht.swest.nhs.uk Fri Feb 3 09:33:48 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Fri Feb 3 09:34:03 2006 Subject: [Histonet] What are peoples feelings on pre lablingslides? Message-ID: That's an interesting response. In my last but one job in London they matched blocks and slides before sending out; in my last job I ran two Cell Path Labs on two Sites and one did and the other didn't. What I was also interested in was the Q/C step as I felt that they only looked at the staining and cutting, but not that enough levels had been taken (the lesion was sampled) or that any further work, including ICC, had been undertaken. I am keen that BMS/ Histotechs push the envelope of their responsibilities by increasing their training and development. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Bonnie Whitaker [mailto:bwhitaker@brownpathology.com] Sent: Friday, February 03, 2006 3:27 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] What are peoples feelings on pre lablingslides? IMHO, ideally, someone is matching blocks and slides before they go out (part of the QA process). I know that it doesn't ALWAYS happen, but I think that regardless of when the slides are labeled, checking them is a prudent thing to do. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vicki Gauch Sent: Friday, February 03, 2006 7:18 AM To: Histonet@lists.utsouthwestern.edu; petepath@yahoo.com Subject: Re: [Histonet] What are peoples feelings on pre lablingslides? We do not pre-label any of our slides in the lab. We cut our slides and then label them as we do each one. As for frozens, we label the slide as we cut the frozen- if we do multiple frozens on that block we label each one as we go. It has been our policy to do it this way for a long time now..in the past we had a few techs who decided to do it "their" way instead of following our procedures and it almost resulted in a case being sent out mislabeled (fortunately it ws caught before it left the lab) which is why we do not allow that practice in our lab. Vicki Gauch AMCH Albany, NY >>> "Stephen Peters M.D." 2/3/2006 8:02:01 AM >>> I am curious to see if it is considered acceptable practice to pre-label multiple slides before cutting the blocks and picking the tissues up on these pre-labeled slides. We came close to a dangerous misdiagnosis because a tech picked up a malignant section from a " part 2 breast biopsy" on a prelabled part 1 slide. Luckily it made no sense that only one of many slides contained tumor that looked like it was coming from an advanced tumor. After playing match the blocks it was obvious that the malignant part one slide matched a part 2 block. It seems to me that this is a potentially dangerous habit despite the convenience of assembly line labeling. Early in my career I stopped labeling my frozen section slides up front and wait until after I pick up the section. When I am cutting frozens I make variable #s of slides depending on the situation. Working quickly under the pressure of multiple cases it is not hard to pick up the wrong slide and make this mistake. I am curious to hear peoples thoughts. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------- CONFIDENTIALITY NOTICE: This email and any attachments may contain confidential information that is protected by law and is for the sole use of the individuals or entities to which it is addressed. If you are not the intended recipient, please notify the sender by replying to this email and destroying all copies of the communication and attachments. Further use, disclosure, copying, distribution of, or reliance upon the contents of this email and attachments is strictly prohibited. To contact Albany Medical Center, or for a copy of our privacy practices, please visit us on the Internet at www.amc.edu. _______________________________________________ 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 From gcallis <@t> montana.edu Fri Feb 3 09:39:54 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Feb 3 09:40:09 2006 Subject: Hardness of tissues RE: [Histonet] processing In-Reply-To: References: Message-ID: <6.0.0.22.1.20060203083424.01b2ed70@gemini.msu.montana.edu> Not an intellectual battle, just a discussion. I would like to share this message from a lady has vast experience working animal tissues, sent yesterday. NOT all animal tissues, obviously, are going to have less % of fat, but some are worse than others. >Jerry Fredenburg did a wonderful workshop back in the dark ages about 20 >yrs ago on the chemistry of processing and staining and talked about bound >water. Funny you should mention it because I was discussing this with my >student only a few days ago. I'm always meaning to dig out that handout >where he explained it so well. They always think I'm making up the term >"bound water". >I think the animals eat healthier and there are less lipids in their >tissue. When I get rodents who were given a fatty diet their tissue are >not so dried out. Their livers are dripping with Oil Red O when I do that >stain on frozens - so much that sometimes coverslipping pushes out the red >stained lipids. > >Andi > At 01:53 AM 2/3/2006, you wrote: >The hardness caused by processing is presently an intellectual battle >between myself and Gayle (covert to this List). > >I'm presently in think mode on this. > >Kemlo Rogerson >Pathology Manager >Ext 3311 >DD 01934 647057 >Mob 07749 754194 > Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From pmarcum <@t> vet.upenn.edu Fri Feb 3 09:53:57 2006 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Fri Feb 3 09:54:12 2006 Subject: Hardness of tissues RE: [Histonet] processing In-Reply-To: <6.0.0.22.1.20060203083424.01b2ed70@gemini.msu.montana.edu> References: <6.0.0.22.1.20060203083424.01b2ed70@gemini.msu.montana.edu> Message-ID: <6.1.1.1.2.20060203104712.019a1bf8@mail.vet.upenn.edu> Ada Feldman also discusses bound water in one of her lectures. I think it is the one about Histology and the Kitchen. I believe Anatech also did a newsletter or flyer with some information on bound water and processing/dehydration etc. Ethel Macrea has also discussed it in the past. It was very interesting and helpful. Animals do appear to be more lean and the lack of fat or lipid content can effect processing. It is why we in the VIR area have very different protocols from those doing human work routinely. We see it here all the time. No Andi you are not making it up and it would be very helpful if someone did more on this subject. Pam Marcum At 10:39 AM 2/3/2006, Gayle Callis wrote: >Not an intellectual battle, just a discussion. > >I would like to share this message from a lady has vast experience >working animal tissues, sent yesterday. NOT all animal tissues, >obviously, are going to have less % of fat, but some are worse than others. > >>Jerry Fredenburg did a wonderful workshop back in the dark ages about 20 >>yrs ago on the chemistry of processing and staining and talked about >>bound water. Funny you should mention it because I was discussing this >>with my student only a few days ago. I'm always meaning to dig out that >>handout where he explained it so well. They always think I'm making up >>the term "bound water". >>I think the animals eat healthier and there are less lipids in their >>tissue. When I get rodents who were given a fatty diet their tissue are >>not so dried out. Their livers are dripping with Oil Red O when I do that >>stain on frozens - so much that sometimes coverslipping pushes out the >>red stained lipids. >> >>Andi >At 01:53 AM 2/3/2006, you wrote: >>The hardness caused by processing is presently an intellectual battle >>between myself and Gayle (covert to this List). >> >>I'm presently in think mode on this. >> >>Kemlo Rogerson >>Pathology Manager >>Ext 3311 >>DD 01934 647057 >>Mob 07749 754194 > >Gayle Callis >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 >406 994-4303 (FAX) > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From laurie.colbert <@t> huntingtonhospital.com Fri Feb 3 10:30:45 2006 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Feb 3 10:30:57 2006 Subject: [Histonet] What are peoples feelings on pre labling slides? Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628006307C95@EXCHANGE1.huntingtonhospital.com> I worked at a hospital for years where we pre-made our slides. Currently, we do not pre make them. I don't feel that there were any more mistakes made when the slides were made out ahead of time. Personally, I make out all my slides for one tray of blocks before I start cutting. I check my block number and slide number before and after picking up each section. I feel that making the change from one method to the other will result in more mistakes at first, but the techs need to always check and double-check their numbering, and once they get used to the new method it shouldn't be an issue. Laurie Colbert -----Original Message----- From: Stephen Peters M.D. [mailto:petepath@yahoo.com] Sent: Friday, February 03, 2006 5:02 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] What are peoples feelings on pre labling slides? I am curious to see if it is considered acceptable practice to pre-label multiple slides before cutting the blocks and picking the tissues up on these pre-labeled slides. We came close to a dangerous misdiagnosis because a tech picked up a malignant section from a " part 2 breast biopsy" on a prelabled part 1 slide. Luckily it made no sense that only one of many slides contained tumor that looked like it was coming from an advanced tumor. After playing match the blocks it was obvious that the malignant part one slide matched a part 2 block. It seems to me that this is a potentially dangerous habit despite the convenience of assembly line labeling. Early in my career I stopped labeling my frozen section slides up front and wait until after I pick up the section. When I am cutting frozens I make variable #s of slides depending on the situation. Working quickly under the pressure of multiple cases it is not hard to pick up the wrong slide and make this mistake. I am curious to hear peoples thoughts. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cindy38017 <@t> aol.com Fri Feb 3 10:41:50 2006 From: cindy38017 <@t> aol.com (cindy38017@aol.com) Date: Fri Feb 3 10:42:12 2006 Subject: [Histonet] New email address Message-ID: <8C7F6F7DD7FC2DC-1104-3BAC@mblk-d37.sysops.aol.com> My new email address is cindy38017@yahoo.com From petepath <@t> yahoo.com Fri Feb 3 10:51:46 2006 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Feb 3 10:51:55 2006 Subject: [Histonet] What are peoples feelings on pre lablingslides? Message-ID: <20060203165147.93469.qmail@web30413.mail.mud.yahoo.com> I want to thank all who responded to my question. These varying opinions all make good points and seem quite valid. This is another example of the variability in in peoples methods that exist in the field of histotechnology. From a medico-legal standpoint I think it would be valuable if practices such as these were better defined so one could properly defend oneself when such a mistake finds it's way to the court room. In this situation of pre-labeling I will suggest to my gang that they should only lable multiple slides for one specimen at a time especially if the two specimens are of the same tissue. If nothing else this reminds us of a potential sourse of error and as Rene and others mentioned the responsibility of the tech in complying with QC measures and double checking these critical steps. Thanks again to all. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From info <@t> instrumedics.com Fri Feb 3 10:57:07 2006 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Feb 3 10:57:31 2006 Subject: [Histonet] histobath Message-ID: <00bb01c628e2$e8540c20$6401a8c0@INSTRUMEDICS22> Bob, You would have a much colder and less toxic system if you switched to the Gentle Jane method which uses a cup of LN2 to chill a heat extractor that is then placed on the Gentle Jane device. It drops at a controlled rate and contacts the tissue first which is uppermost on the mound of CryoGel. The tissue and the gel freezes in 10 -15 seconds at a much lower temperature (with thermal exchange)than in the Histobath and minimizes ice crystal artifact. See the details on our web site www.instrumedics.com Bernice From anh2006 <@t> med.cornell.edu Fri Feb 3 11:37:05 2006 From: anh2006 <@t> med.cornell.edu (Andrea Hooper) Date: Fri Feb 3 11:37:16 2006 Subject: [Histonet] What are peoples feelings on pre labling slides? Message-ID: I believe this to be entirely histotech dependent. I have worked with some histologists who never had a problem as their attention to detail was high ... and I have also worked with others who would constantly confuse slides if labeled ahead of time. Ideally we are all attentive to detail and can handle labeling ahead of time, but practically in the real world that just isn't the case. Everyone is different. I believe a supervisor needs to know their staff well enough to enforce the proper criteria to achieve NO MISTAKES! -----Original Message----- From: Stephen Peters M.D. [petepath@yahoo.com] Sent: Friday, February 03, 2006 5:02 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] What are peoples feelings on pre labling slides? I am curious to see if it is considered acceptable practice to pre-label multiple ? slides before cutting the blocks and picking the tissues up on these pre-labeled ? slides. We came close to a dangerous misdiagnosis because a tech picked up ?? a malignant section from a " part 2 breast biopsy" on a prelabled part 1 slide. Luckily it made no sense that only one of many slides contained tumor that ? looked like it was coming from an advanced tumor. After playing match the blocks ?? it was obvious that the malignant part one slide matched a part 2 block. It seems ? to me that this is a potentially dangerous habit despite the convenience of ? assembly line labeling. Early in my career I stopped labeling my frozen section slides ? up front and wait until after I pick up the section. When I am cutting frozens I? make ?? variable #s of slides depending on the situation. Working quickly under the ?? pressure of multiple cases it is not hard to pick up the wrong slide and make ?? this mistake. I am curious to hear peoples thoughts. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 ? From PMonfils <@t> Lifespan.org Fri Feb 3 12:15:48 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Feb 3 12:15:58 2006 Subject: [Histonet] collecting fresh tissue prior to perfusion Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717660@lsexch.lsmaster.lifespan.org> Caroline, I have done what you describe on a few occasions, not because I wanted to do DNA analysis but because I wanted to do electron microscopy along with standard histology. Cutting off a quarter of a lobe would probably allow excessive leakage during subsequent perfusion, not only because of the surface area involved, but also because you would undoubtedly cut some fairly large vessels. I don't know how much tissue volume you need for DNA studies, but we needed only a tiny biopsy for EM (a couple of cubic millimeters). We cut a tiny notch from the edge of one lobe, using a small scalpel, and the amount of capillary leakage from such a small peripheral incision was negligible. Paul Monfils > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Caroline Bass > Sent: Wednesday, February 1, 2006 10:52 AM > To: Histonet > Subject: [Histonet] collecting fresh tissue prior to perfusion > > Hello, > > I have what may be a silly question but I thought maybe it is worth a > try. I am in a situation where the best way to collect liver tissue > for my purposes is to perfuse the mouse with PFA. However, on > occasion I need a small bit of fresh liver tissue to analyze for > DNA. I was wondering if it is possible to collect a portion of a > liver lobe at some point in the perfusion process before I use > fixative? For example, I could start the perfusion with heparinized > saline, clip a quarter of one lobe of the liver off and then switch > over to the PFA. I would then proceed with the perfusion as normal. > > The only complication I can think of is that the loss of a > significant portion of the liver could effect how well the rest of > the liver perfuses. > > Has anyone tried this or is this something I should avoid? I am > trying to cut down on the number of mice I am using. > > My goal is to obtain a completely fixed liver lobe that I can section > all the way through. It is my understanding that the mouse liver is > too large to fix by immersion in PFA, so it either has to be cut into > blocks or the mouse perfused. > > Thanks, > > Caroline > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From gu.lang <@t> gmx.at Fri Feb 3 12:40:12 2006 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Feb 3 12:40:16 2006 Subject: AW: [Histonet] Re-processing (Johnson) In-Reply-To: Message-ID: <000101c628f1$446ab0f0$eeeea8c0@SERVER01> What kind of processor is used with this method? We have a VIP and were instructed to take care of too much paraffin in the container when starting the run (particels left after cleaning). So it sounds very easy to reprocess the tissue, but I don't want to break down the machine. Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Tony Henwood Gesendet: Donnerstag, 02. Februar 2006 23:23 An: Celebre Julia; Histonet@lists.utsouthwestern.edu Betreff: RE: [Histonet] Re-processing The following technique from Johnson (2003) Histologic 36(1):21-22 works very well: While the tissue is still hot (ie the wax is still molten) blot the tissue dry, place it back in its cassette and place the cassette in 10% formalin for reprocessing. I have often had recourse to use this method and have found it to dramatically improve the results in at least 90% of cases. The excellent results are probably due to the protective nature of the wax present in the adequately processed portions of the block. This insulates the tissue from the harmful effects of ethanol on the adequately processed portions of the tissue preventing the tissue from becoming hard and brittle (Johnson 2003). Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Celebre Julia Sent: Friday, 3 February 2006 4:21 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Re-processing Calling all processing and reprocessing experts out there.... I'm sure all labs over the years have run into processing troubles ranging from instrument malfunctions during the night to misplacement of solutions (usually a water before the xylene), causing pure horror for the technologist who opens up the retort in the morning. It's sad to say that our department is experienced in these situations, and everytime something happens to the processing it's always difficult to decide how best to reprocess the sections. Each time we've reprocessed our sections (usually due to the introduction of water before the xylene or paraffin stations) on shortened absolute alcohol, xylenes and paraffin times, they'v ended up being extremely hard and brittle, which does not make our pathologist happy.... Is there a way of reprocessing tissue without damaging it? Would adding a softener to the alcohols reduce the hardening affects of reprocessing? How much time can be safely shaved off if the tissues have already been through a regular program? Any help is greatly appreciated, Julia Celebre MLT Anatomic Pathology Hamilton General Hospital Hamilton, Ontario Canada 905-527-0271 ext 46179 email: celebrej@hhsc.ca This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lcastillos <@t> icoria.com Fri Feb 3 13:04:27 2006 From: lcastillos <@t> icoria.com (Castillos, Luminita) Date: Fri Feb 3 13:04:37 2006 Subject: [Histonet] LCM & Immunostaining for macrophages Message-ID: Hi, I am doing LCM and my project is to capture pure population of macrophages from synovial tissue, from FFPE slides. I am looking for an immunostaining method for macrophages but to be feasible for LCM work. I will appreciate any advice for my problem. Thank you so much. Regards, Luminita Luminita Castillos, Ph.D. Research Scientist Icoria, Inc., A Clinical Data Inc. company 108 T.W. Alexander Drive P.O. Box 14528 RTP, NC 27709-4528 Phone: 919-425-2967 Cell: 919-302-0485 www.icoria.com lcastillos@icoria.com Effective Dec. 21, 2005, Icoria became a Clinical Data, Inc. company, the worldwide leader in the molecular diagnostic field. The Pharmacogenomics and the Molecular Services Division of Clinical Data, Inc. (now with expanded genomics offerings from Icoria) specializes in the development and commercialization of molecular biology and pharmacogenomics solutions to biotechnology and pharmaceutical companies, academia and government institutions. From sharon.osborn <@t> dnax.org Fri Feb 3 16:11:19 2006 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Feb 3 16:11:52 2006 Subject: [Histonet] RE: pre-labeling slides Message-ID: <29B25753F6B1D51196110002A589D444032C49EC@PALMSG30.us.schp.com> Some people's handwriting leaves room for misinterpretation or no interpretation at all. I have worked in situations with performing the hand labeling at time of cuttng the block; pre-labeling the slides the day before from the blocks made; using the automated slide labelers, etc. Due to the handwriting problem, smearing of the labeling, etc. I prefer the machine labeled slides. These can also serve as the permanent label since a paper label is not needed. Plus, the machine labeled slides are another way to decrease the repetitive motion problems we histology types are prone to due to the nature of our work. As most people have pointed out, it is up to the tech to double check the slide and the block. There can still be mis-labeled slides when done at the time of the cutting by transposing the numbers, sloppy penmanship, etc. It seems to be inherent in humans that some mistakes will be made in the repetitive work such as this. Some labs have the slides and blocks matched again before sending out. This is a time consuming method, yes; however, it can provide that extra measure of assurance that the slides are for the correct cases, etc. Perhaps having a percent of error for X number of specimens is to be considered appropriate. I realize that some techs/supervisors will disagree with me on this...we need to provide a comfort level that techs are not uptight such there is increased chance of labeling error due to the pressure to be "perfect". Knowing there is a check system of some type in place can alleviate this and provide the extra measure of "law suit proofing" for the industry. Sharon Osborn DNAX, SP BioPharma Palo Alto, CA ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From djamesnz <@t> orcon.net.nz Fri Feb 3 16:13:35 2006 From: djamesnz <@t> orcon.net.nz (Darren James) Date: Fri Feb 3 16:14:04 2006 Subject: [Histonet] What are peoples feelings on pre labling slides? In-Reply-To: Message-ID: <000501c6290f$14cceb60$0301010a@DAZZA> I have worked in labs where all variations have been used. I found no more or less mistakes in any of the sites. I personally prefer pre labelling. What works for one person may not work for another. Some people have excellent recall of numbers and attention spans, some do not. I think that at the end of the day there will be mistakes what ever method is used, we are all human after all (well most of us) but it is all about having adequate systems in place to minimise the impact of these mistakes. Have a great day. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Andrea Hooper Sent: Saturday, 4 February 2006 6:37 a.m. To: Histonet (E-mail) Cc: Stephen Peters M.D. Subject: Re: RE: [Histonet] What are peoples feelings on pre labling slides? I believe this to be entirely histotech dependent. I have worked with some histologists who never had a problem as their attention to detail was high ... and I have also worked with others who would constantly confuse slides if labeled ahead of time. Ideally we are all attentive to detail and can handle labeling ahead of time, but practically in the real world that just isn't the case. Everyone is different. I believe a supervisor needs to know their staff well enough to enforce the proper criteria to achieve NO MISTAKES! -----Original Message----- From: Stephen Peters M.D. [petepath@yahoo.com] Sent: Friday, February 03, 2006 5:02 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] What are peoples feelings on pre labling slides? I am curious to see if it is considered acceptable practice to pre-label multiple ? slides before cutting the blocks and picking the tissues up on these pre-labeled ? slides. We came close to a dangerous misdiagnosis because a tech picked up ?? a malignant section from a " part 2 breast biopsy" on a prelabled part 1 slide. Luckily it made no sense that only one of many slides contained tumor that ? looked like it was coming from an advanced tumor. After playing match the blocks ?? it was obvious that the malignant part one slide matched a part 2 block. It seems ? to me that this is a potentially dangerous habit despite the convenience of ? assembly line labeling. Early in my career I stopped labeling my frozen section slides ? up front and wait until after I pick up the section. When I am cutting frozens I? make ?? variable #s of slides depending on the situation. Working quickly under the ?? pressure of multiple cases it is not hard to pick up the wrong slide and make ?? this mistake. I am curious to hear peoples thoughts. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 ? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From n.aljehani <@t> btinternet.com Sun Feb 5 04:53:30 2006 From: n.aljehani <@t> btinternet.com (Dr. Nami Aljehani) Date: Sun Feb 5 04:53:41 2006 Subject: [Histonet] Histotechnologist Job Opportunity Message-ID: <20060205105330.55363.qmail@web86704.mail.ukl.yahoo.com> Histotechnologist Job Opportunity King Fahad Medical City (KFMC) in Riyadh, Saudi Arabia is newly opened medical facility and considered the largest medical facility in the Middle East that specializes in treating rare and specialty diseases (e.g. Oncology, Neurology, Genetic diseases, Cardiac diseases). The complex incorporates 4 hospitals and a medical school. The capacity of this medical city is up to 1200 bed and expected to treat more than 50,000 in-house patients and more than 600,000 outpatients annually. Immediate job opportunities for 2 Histotechnologist positions in the Anatomical pathology and Cytology Department The laboratory is equipped with most of the equipment that is needed to run a modern pathology lab. However, there are some machines that will be arriving in the near future such as the Autoimmune stainer. I'm the technical supervisor with 2 senior Histopathology Technologist and 4 Junior Medical Technologist. Currently the work load is not that heavy but after the Medical City is up and running at full capacity we will be extremely busy. Job Description Maintain a high standard of accuracy and technical competence in the performance of the daily routine procedures, special stains, immunohistochemical stains, and frozen sections. Participates in scheduled, regularly allocated on call duty and flexible scheduling to permit required services, e.g. on weekends or holidays. Participates in continuing education activities and proficiency testing as required or recommended by accrediting agencies. Participates in and contributes to quality assurance (performance improvement) programs as mandated by the Department, the institution and regulatory and accrediting agencies. Performs other relevant duties as requested by immediate supervisor or section chief in the operation of the histology laboratory. Possesses excellent communication and understanding of multi cultural environment. Qualifications: 1. Bachelors degree in Medical Technology or science chemistry and biology courses 2. ASCP Histotechnology (HT) certification or equivalent 3. Five years of experience in an accredited Pathology department 4. Experience with medical photography desirable 5. Experience in Molecular biology techniques desirable 6. Basic acquaintance with computer. If interested, please forward resume to Ms. Lora Wood Head, International Recruitment Unit Telephone: +966 1 465 6666 ext. 8156, 8003 or 2023 Direct Fax Line: +966 1 416 1158 OR Dr Nami Aljehani, PhD, FRMS, FISAC Histopathology and Cytology Supervisor King Fahad Medical City, Riyadh Work Tel # + 966-1-4656666 Ext. 1537 Bleep 3289 Mobile # +966-506544-275 "Success is a state of mind. If you want success, start thinking of yourself as a success." --Joyce Brothers --------------------------------- Yahoo! Messenger NEW - crystal clear PC to PC calling worldwide with voicemail From kemlo <@t> f2s.com Sun Feb 5 10:11:28 2006 From: kemlo <@t> f2s.com (kemlo) Date: Sun Feb 5 10:11:40 2006 Subject: [Histonet] overprocessing Message-ID: <200602051611.k15GBSVT012696@outmail.freedom2surf.net> Formaldehyde fixes we assume by creating cross links between proteins thereby creating a gel; soluble proteins are bound and that gives strength to the gel and helps to withstand processing. These cross links are between basic acid lysine and other groups (amido, peptide, hydroxyl, etc). Formalin is not a coagulant (precipitant) fixative but it's an additive fixative; it does not harden and is reversible; however it stabilises the proteins and prevents subsequent hardening by alcohols. Ethanol disrupts the hydrophobic bonds which maintain the structure of proteins (coagulant fixative); hydrogen bonds are more stable in ethanol than water and so the structure is preserved. Both bound and unbound water molecules will be replaced during fixation. I can only conclude that formalin is unable to prevent the damaging effects of processing if the gel has not been allowed to form adequately or if it the gel is disrupted by 'washing'. I also assume that too much 'processing' collapses the gel (removes soluble proteins) and allow ethanol to 'harden' proteins (look at the bottom of the processing retort or the waste); maybe this collapse is in part due to the removal of 'bound' water or the condensation/ removal of proteins. Different tissues or different species contain differing proteins and therefore the effects of fixation and processing will differ to. The effects of raised temperatures will be to exacerbate the condensation/ removal of proteins from the gel or to increase the deleterious effects of ethanol on 'unfixed' tissues (increase its rate of action). I don't follow the illogical presumption that fat has anything to do with it as the effects would not only differ between species but would differ within species (fat one's would process differently than thin one's, in any given species). It may well be that the difference is the result of the different proteins that make up tissue in the differing species and the vagaries of the stabilising effects of formalin which include its removal by processing. IMHO Kemlo From billingconsultants <@t> yahoo.com Sun Feb 5 15:07:01 2006 From: billingconsultants <@t> yahoo.com (Billing Consultants, LLC) Date: Sun Feb 5 15:07:10 2006 Subject: [Histonet] Attn Vendors Message-ID: <20060205210701.61250.qmail@web54215.mail.yahoo.com> Hi Everyone, I would like to obtain general pricing information on new histology equipment for a client interested in setting up a histology lab. The equipment they are interested in are as follows: Basic Microtome, non-automated Closed system Tissue Processor, up to 100 blocks capacity Cryostat Embedding Center Waterbath Basic automated H&E stainer I am assisting the client in obtaining ballpark figures and contact information for the various vendor options. The client will then contact each vendor directly when the project proceeds beyond this initial stage. Thank you in advance for your assistance. Louri Roberts --------------------------------- Yahoo! Mail - Helps protect you from nasty viruses. From carl.hobbs <@t> kcl.ac.uk Sun Feb 5 15:40:21 2006 From: carl.hobbs <@t> kcl.ac.uk (carlos) Date: Sun Feb 5 15:43:51 2006 Subject: [Histonet] Re: overprocessing Message-ID: <001a01c62a9d$1f9e06c0$0201a8c0@carlos> Hi Kemlo, I must disagree re hardening......if only in my experience of the following: I have processed gelatin ( 1%)blocks after formalin fixation ( 1cm blocks fixed for 2, 5, 10 and 60 days) and find that they all suffer from shrinkage and hardening. My processing is via gradual alcohols to alc/xylene 1:1, then xylene x2 and wax x2. Two hrs in each. Be grateful for enlightenment. Best wishes carl From AnthonyH <@t> chw.edu.au Sun Feb 5 17:01:03 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Sun Feb 5 17:01:20 2006 Subject: [Histonet] Re-processing (Johnson) Message-ID: We use Leica carousel processors. I would not expect too much wax carryover in the VIP using this method Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Saturday, 4 February 2006 5:40 AM To: Histonetliste (Histonetliste) Subject: AW: [Histonet] Re-processing (Johnson) What kind of processor is used with this method? We have a VIP and were instructed to take care of too much paraffin in the container when starting the run (particels left after cleaning). So it sounds very easy to reprocess the tissue, but I don't want to break down the machine. Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Tony Henwood Gesendet: Donnerstag, 02. Februar 2006 23:23 An: Celebre Julia; Histonet@lists.utsouthwestern.edu Betreff: RE: [Histonet] Re-processing The following technique from Johnson (2003) Histologic 36(1):21-22 works very well: While the tissue is still hot (ie the wax is still molten) blot the tissue dry, place it back in its cassette and place the cassette in 10% formalin for reprocessing. I have often had recourse to use this method and have found it to dramatically improve the results in at least 90% of cases. The excellent results are probably due to the protective nature of the wax present in the adequately processed portions of the block. This insulates the tissue from the harmful effects of ethanol on the adequately processed portions of the tissue preventing the tissue from becoming hard and brittle (Johnson 2003). Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Celebre Julia Sent: Friday, 3 February 2006 4:21 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Re-processing Calling all processing and reprocessing experts out there.... I'm sure all labs over the years have run into processing troubles ranging from instrument malfunctions during the night to misplacement of solutions (usually a water before the xylene), causing pure horror for the technologist who opens up the retort in the morning. It's sad to say that our department is experienced in these situations, and everytime something happens to the processing it's always difficult to decide how best to reprocess the sections. Each time we've reprocessed our sections (usually due to the introduction of water before the xylene or paraffin stations) on shortened absolute alcohol, xylenes and paraffin times, they'v ended up being extremely hard and brittle, which does not make our pathologist happy.... Is there a way of reprocessing tissue without damaging it? Would adding a softener to the alcohols reduce the hardening affects of reprocessing? How much time can be safely shaved off if the tissues have already been through a regular program? Any help is greatly appreciated, Julia Celebre MLT Anatomic Pathology Hamilton General Hospital Hamilton, Ontario Canada 905-527-0271 ext 46179 email: celebrej@hhsc.ca This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ 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 From hornd <@t> uthscsa.edu Sun Feb 5 22:19:09 2006 From: hornd <@t> uthscsa.edu (Horn, Diane A) Date: Sun Feb 5 22:19:13 2006 Subject: [Histonet] Immunohistochemistry on mouse teeth Message-ID: <819189266B78D543A02D9D3956EE4F6C030CEC99@addax.win.uthscsa.edu> I am currently doing IHC on develping mouse teeth and I am getting only backgroung staining. The tissues are fixed O/N in 4% paraformaldehyde @4 degrees and embedded in paraffin. I have cut my sections @ 4um. I have tried HIER with Vector unmasking solution (boiling) for 2 minutes and using a kit for PCNA from Zymed. Has anyone done IHC on teeth that could give me some advice on what to try next? Thanks! Diane From jkiernan <@t> uwo.ca Sun Feb 5 23:23:15 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Sun Feb 5 23:22:37 2006 Subject: [Histonet] Re: overprocessing References: <001a01c62a9d$1f9e06c0$0201a8c0@carlos> Message-ID: <43E6DD43.9650D879@uwo.ca> Dear Carlos (whoever and wherever you are), You are quite right, and your observations are supported by published literature that has been through the academic (= accept nothing without proof) peer-review process. Hardness is difficult to evaluate, but anyone can measure changes in volume or linear dimensions. This was all done decades ago, and I'll be happy to send literature references to anyone who's seriously interested. Cross-linked polymers (such as the gels used in PAGE) shrink rather suddenly when water removal (by evaporation or alcohol) hits a critical level. I tried this in the lab after reading a Scientific American article about 20 years ago. There was a rapid change from soft & slimy to hard & gritty, exactly as Sci. Am. said it would be. I can't find the ref for this pre-computers Sci. Am. article. Shrinkage of animal tissues is less than that of highly hydrated polyacrylamide gels or gelatin. A linear measurement in a stained and mounted paraffin section may be about 70% of the in vivo measurement, but the % can vary with the organ. I can send references to anyone who's intersted in following this up. John Kiernan London, Canada _______________________________ carlos wrote: > > Hi Kemlo, > I must disagree re hardening......if only in my experience of the following: > I have processed gelatin ( 1%)blocks after formalin fixation ( 1cm blocks > fixed for 2, 5, 10 and 60 days) and find that they all suffer from shrinkage > and hardening. My processing is via gradual alcohols to alc/xylene 1:1, then > xylene x2 and wax x2. Two hrs in each. > Be grateful for enlightenment. > Best wishes > carl > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Sun Feb 5 23:23:53 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Sun Feb 5 23:23:13 2006 Subject: [Histonet] Immunohistochemistry on mouse teeth References: <819189266B78D543A02D9D3956EE4F6C030CEC99@addax.win.uthscsa.edu> Message-ID: <43E6DD69.F556DEE0@uwo.ca> One possibility is that the sections do not contain the antigen you are trying to detect. Do you have simultaneously processed known positive control sections? John Kiernan Anatomy, UWO London, Canada. ------------------------------ "Horn, Diane A" wrote: > > > I am currently doing IHC on develping mouse teeth and I am getting only backgroung staining. The tissues are fixed O/N in 4% paraformaldehyde @4 degrees and embedded in paraffin. I have cut my sections @ 4um. I have tried HIER with Vector unmasking solution (boiling) for 2 minutes and using a kit for PCNA from Zymed. Has anyone done IHC on teeth that could give me some advice on what to try next? > > Thanks! > Diane > From carl.hobbs <@t> kcl.ac.uk Mon Feb 6 01:57:52 2006 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Mon Feb 6 01:58:24 2006 Subject: [Histonet] re: overprocessing Message-ID: <003301c62af3$07bcc8b0$112b5c9f@Carlos> Thank you for your comments, John. My first post was under a guy called Ted Brain who was always looking into processing /embeding protocols and reagents in an attempt to find the "optimal " for our bone and teeth! It was years ago when I taught Histopathology that I would use , in a simple ( OK, plagiarised) practical on "The effects of Fixation" , 1cm cubes of gelatin, fixed using a range of coagulant/additive fixing fluids to demonstrate the penetration rates and the physical effects that the different fixing agents had on pure protein. A qualitative assessment only....... wouldn't expect more from students at 7.30pm after they'd been at college since 10am. Regarding my initial remark: I didn't notice any shrinkage difference between the short-time fixed gelatin blocks and the ones that had been fixed for ages. But then again, there is the effect of alcohol in the processor which is significant. I wasn't doing this comparison under rigid exptl conditions: we have many perfused- formalin fixed specimens which have been embedded in gelatin, vibratome-cut and that I then wished to process to pwax. Would extended fixation time of the gelatin "stiffen" it sufficiently to prevent shrinkage? The answer , for me, is "No". So, I take off the gelatin, before processing. No "exact science" there, Rene. Sorry! Carl From esther.peters <@t> verizon.net Mon Feb 6 07:33:17 2006 From: esther.peters <@t> verizon.net (Esther Peters) Date: Mon Feb 6 07:33:15 2006 Subject: [Histonet] Re: overprocessing References: <001a01c62a9d$1f9e06c0$0201a8c0@carlos> <43E6DD43.9650D879@uwo.ca> Message-ID: <43E7501D.6050908@verizon.net> Dear John, Thank you for your insights and I would appreciate getting the references. We have enrobed specimens in 1.5% low gelling temperature agarose. While sometimes we have no problems with our short processing runs, sometimes the agarose shrinks and hardens. I pre-embedded bacteria in the agarose to form 1-2 mm-thick disks, then processed and embedded on biopsy times. About half the samples shrank. I had one cassette containing two disks, these were from one that had been sliced in half, so otherwise both were treated the same. One disk shrank and hardened, the other one was fine! So there is a very narrow threshold at which the water removal goes "critical." I have not worked with HistoGel. Is this a better product for this purpose compared to agarose? Esther Peters, Ph.D. George Mason University John Kiernan wrote: > Dear Carlos (whoever and wherever you are), > > You are quite right, and your observations are > supported by published literature that has been > through the academic (= accept nothing without > proof) peer-review process. > > Hardness is difficult to evaluate, but anyone can > measure changes in volume or linear dimensions. > This was all done decades ago, and I'll be happy > to send literature references to anyone who's > seriously interested. > > Cross-linked polymers > (such as the gels used in PAGE) shrink rather > suddenly when water removal (by evaporation or > alcohol) hits a critical level. I tried this in > the lab after reading a Scientific American > article about 20 years ago. There was a rapid > change from soft & slimy to hard & gritty, exactly > as Sci. Am. said it would be. I can't find the ref > for this pre-computers Sci. Am. article. > > Shrinkage of animal tissues is less than that of > highly hydrated polyacrylamide gels or gelatin. A > linear measurement in a stained and mounted > paraffin section may be about 70% of the in vivo > measurement, but the % can vary with the organ. > > I can send references to anyone who's intersted in > following this up. > > John Kiernan > London, Canada > _______________________________ > carlos wrote: > >>Hi Kemlo, >>I must disagree re hardening......if only in my experience of the following: >>I have processed gelatin ( 1%)blocks after formalin fixation ( 1cm blocks >>fixed for 2, 5, 10 and 60 days) and find that they all suffer from shrinkage >>and hardening. My processing is via gradual alcohols to alc/xylene 1:1, then >>xylene x2 and wax x2. Two hrs in each. >>Be grateful for enlightenment. >>Best wishes >>carl >> >>_______________________________________________ >>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 > > From b003046 <@t> nf.au.dk Mon Feb 6 07:54:16 2006 From: b003046 <@t> nf.au.dk (b003046@nf.au.dk) Date: Mon Feb 6 07:54:27 2006 Subject: [Histonet] Negative control for rabbit IgG Message-ID: <1139234056.43e755083f1e5@webmail.nf.au.dk> Hi, I am trying to get the DAKO polyclonal rabbit anti-human c-kit (CD117) to work in rats in paraffin. At this moment I get nice results but to be completely sure that it is positiv I need to get my negative control to work. I am using a negativ control for rabbit IgG, (DAKO;X0903) which gives me a lot of non-specific binding. Do anybody have a suggestion for another negative control I could use? Thanks, Mette K. Hagensen Department of Cardiology B, Research Unit Aarhus University Hospital, Skejby Hospital Brendstrupgaardsvej 100 8200 Aarhus N Denmark Phone: +45 89496237 Mail: b003046@nf.au.dk From petepath <@t> yahoo.com Mon Feb 6 08:18:42 2006 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Mon Feb 6 08:18:55 2006 Subject: [Histonet] Avoiding bilaterality mistakes Message-ID: <20060206141842.63935.qmail@web30408.mail.mud.yahoo.com> After our recent near miss on a bilateral breast case ( see fridays post :What are peoples feelings on pre labling slides?) we will be using different colored cassettes and matching slides for each side. I got this idea from Joyce Weems who told me she uses colored agar markers for laterality ( Thanks Joyce). This should help to prevent confusion at the grossing, cutting and reading stages. Thanks to all who answered my first post. I thought I would share what I hope is a good solution. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From DavidH <@t> marketlabinc.com Mon Feb 6 08:35:39 2006 From: DavidH <@t> marketlabinc.com (David Haagsma) Date: Mon Feb 6 08:35:49 2006 Subject: [Histonet] Immunofluorescence Text Message-ID: <19E3602A16438E48B51A4250CA04B5F64A2C07@exchange.marketlab.com> Dear Histonet; We have a customer that is looking for a good immuno book. We currently do not offer a book of this type and I do not have any recommendations for her - can you help her out? Thank you, Dave Haagsma MT(ASCP) MarketLab Inc. Subject: Find Product Form name...........: Caroline Ghaleb suggestion.....: Dear Sir, You have a good number of educational book, unfortunately I'm looking to buy a book about immunofluorescence. I appreciate if you can let me know if you can provide such a book. Thanks a lot. Caroline Ghaleb phone..........: email..........: teknolab@rcn.com From Ronnie_Houston <@t> bshsi.com Mon Feb 6 08:51:34 2006 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Mon Feb 6 08:53:07 2006 Subject: [Histonet] Iron demonstration in smears Message-ID: <00FBB1F3374BE24AAB7ACC6FCC7C617F561F46@bsrexms01.bshsir.com> One of our pathologists is asking how long can smears be left at room temperature before being stained for hemosiderin; he is primarily concerned with bone marrow and urine preps. Thanks ronnie Ronnie Houston Director of Anatomic Pathology Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 2877972 (804) 2877906 - fax ronnie_houston@bshsi.com ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone, and permanently delete the original e-mail, attachment(s), and any copies. From kemlo.rogerson <@t> waht.swest.nhs.uk Mon Feb 6 09:19:59 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Mon Feb 6 09:20:22 2006 Subject: [Histonet] Iron demonstration in smears Message-ID: Obviously if they are to be 'fixed' you don't use a fixative containing acid; if air dried, air dry then fix them in methanol (not Pap fix as it contains glacial acetic acid) and in both cases they ought to last as long as Pap smears, which I guess is months. As long as the proteins are stabilised than everything ought to be Ok; fungus however can be a problem as I've seen Pap smears infested with hyphae after a few months. If you don't store them properly you can get such contamination; interesting Carbo wax in the Pap Fix is supposed to act as a barrier to many things; maybe you could take the acetic acid out? Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Houston, Ronnie [mailto:Ronnie_Houston@bshsi.com] Sent: Monday, February 06, 2006 2:52 PM To: Histonet (E-mail) Subject: [Histonet] Iron demonstration in smears Importance: High One of our pathologists is asking how long can smears be left at room temperature before being stained for hemosiderin; he is primarily concerned with bone marrow and urine preps. Thanks ronnie Ronnie Houston Director of Anatomic Pathology Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 2877972 (804) 2877906 - fax ronnie_houston@bshsi.com ____________________________________________________________________________ ____________________________________________________ ____________________________________________________________________________ ____________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone, and permanently delete the original e-mail, attachment(s), and any copies. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tkngflght <@t> yahoo.com Mon Feb 6 09:41:56 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Mon Feb 6 09:42:05 2006 Subject: [Histonet] Iron demonstration in smears In-Reply-To: <00FBB1F3374BE24AAB7ACC6FCC7C617F561F46@bsrexms01.bshsir.com> Message-ID: <20060206154157.55307.qmail@web50911.mail.yahoo.com> In my experience open time is very long, having stained slides that were years old with consistent results to those stained within days of collection. Staining specificity and 'detail' depends more on your staining method than on storage time. If you make your reagents up in bulk or purchase pre-made, and if you stain at room temp or microwave, your open time will be shorter due to reduced sensitivity of the test method. One group I worked for researched FE++ staining sensitivity for a REALLY long time and found that making the reagents up fresh EACH AND EVERY TIME proved beneficial (weigh out the reagents and store them in sealed dry alliquots for speed on clinical benches) and once constitututed, heat (temp??) in an H2O bath to temp before staining for uniformity, using mixed reagent only once immediately after preparation for one batch. This overcame any storage issues where we could reproduce staining several years after dry-storing squash, smear and touch preps at room temp filed with our archive H&Es. I do not have a copy of the procedure or reference but if anyone is interested I'd be happy to try to find it again. We had another process of spinning down aspirates at a certain speed (2800 rpm I think??) for ten minutes and the spicules layered above the red cells. These were siphoned off and manipulated on skin burn dressing before fixing/blocking to create much cleaner aspirate blocks. One of the pathologist in the group pattented a spin tube with a screw-off bottom just for this but orange topped 10ml slant bottomed tubes folks use to transport RPMS and such work just fine (sealed tops) Hope this helps a bit-- Cheryl Kerry Full Staff Inc. "Houston, Ronnie" wrote: One of our pathologists is asking how long can smears be left at room temperature before being stained for hemosiderin; he is primarily concerned with bone marrow and urine preps. Thanks ronnie Ronnie Houston Director of Anatomic Pathology Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 2877972 (804) 2877906 - fax ronnie_houston@bshsi.com ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone, and permanently delete the original e-mail, attachment(s), and any copies. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Feb 6 10:02:11 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Feb 6 10:02:18 2006 Subject: Color coding Re: [Histonet] Avoiding bilaterality mistakes In-Reply-To: <20060206141842.63935.qmail@web30408.mail.mud.yahoo.com> References: <20060206141842.63935.qmail@web30408.mail.mud.yahoo.com> Message-ID: <6.0.0.22.1.20060206085707.01b64480@gemini.msu.montana.edu> Color coding has bailed us out of problems many, many times. This is an excellent suggestion. I am very impressed at use of automated slide labeling IF specimen volume/workload and budget allows it, it certainly is easier on the hands too. At 07:18 AM 2/6/2006, you wrote: >After our recent near miss on a bilateral breast case ( see fridays post >:What > are peoples feelings on pre labling slides?) we will be using different > colored > cassettes and matching slides for each side. I got this idea from Joyce > Weems who told me she uses colored agar markers for laterality ( Thanks > Joyce). This should > help to prevent confusion at the grossing, cutting and reading stages. > Thanks to all > who answered my first post. I thought I would share what I hope is a > good solution. >Stephen Peters M.D. Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From PMonfils <@t> Lifespan.org Mon Feb 6 11:00:36 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon Feb 6 11:00:49 2006 Subject: [Histonet] Iron demonstration in smears Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717662@lsexch.lsmaster.lifespan.org> In reference to the previously posted comment about fungal growth on stored slides ... This problem is almost always the result of refrigeration. There is a natural tendency to expect better preservation under refrigeration than at room temperature, and this is certainly true for tubes of blood, whole tissue specimens, lunch meat and cheese. But slides - either cut sections or smears - are a different story. Fungi have four basic requirements for optimal growth - a nutrient source, warmth, darkness, and moisture. Moist materials like those mentioned above have to be refrigerated. On the other hand, while slides stored in a slide box at room temperature in a drawer or on a shelf may provide nutrients, warmth and darkness, they are in a dessicated state, and without moisture fungi won't grow. (Of course this might not apply if you work in a very humid climate and don't have air conditioning in the lab). The problem with refrigeration is that it provides the additional growth requirement - moisture; and while the relatively low temperature of a refrigerator will slow the growth of fungi, it won't stop such growth, as anyone realizes who has thrown out a moldy piece of lunch meat or cheese. To stop fungal growth thermally you need sub-freezing temperatures. Therefore the best long-term preservation of dry fixed specimens is at room temperature. Methanol fixation is good, as already stated by another poster, and several types of cytology spray fixatives also work very well. From HornHV <@t> archildrens.org Mon Feb 6 11:07:06 2006 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Mon Feb 6 11:07:30 2006 Subject: Color coding Re: [Histonet] Avoiding bilaterality mistakes Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDF82@EMAIL.archildrens.org> We use color coding too and I presented a poster about this at NSH. We color code all our biopsies. Each biopsy case receives a different color cassette. Those blocks are put on the same color slides as the biopsy case. It does help with labeling errors. We have 6 different colors we alternate using. Most of our blocks are biopsies. This would work for large cases too, like breast resections, colon resections, prostate chips..... Hazel -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Monday, February 06, 2006 10:02 AM To: Stephen Peters M.D.; Histonet@lists.utsouthwestern.edu Subject: Color coding Re: [Histonet] Avoiding bilaterality mistakes Color coding has bailed us out of problems many, many times. This is an excellent suggestion. I am very impressed at use of automated slide labeling IF specimen volume/workload and budget allows it, it certainly is easier on the hands too. At 07:18 AM 2/6/2006, you wrote: >After our recent near miss on a bilateral breast case ( see fridays post >:What > are peoples feelings on pre labling slides?) we will be using different > colored > cassettes and matching slides for each side. I got this idea from Joyce > Weems who told me she uses colored agar markers for laterality ( Thanks > Joyce). This should > help to prevent confusion at the grossing, cutting and reading stages. > Thanks to all > who answered my first post. I thought I would share what I hope is a > good solution. >Stephen Peters M.D. Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From pruegg <@t> ihctech.net Mon Feb 6 11:50:47 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Mon Feb 6 11:50:50 2006 Subject: [Histonet] Immunofluorescence Text In-Reply-To: <19E3602A16438E48B51A4250CA04B5F64A2C07@exchange.marketlab.com> Message-ID: <200602061750.k16HodDZ008025@chip.viawest.net> The NSH IHC Resource Group has put together a list of reference books for IHC. I don't think there is specifically one on just F-IHC but several of them relate. You can join the IHCRG online at www.ihcrg.org for this and many other benefits relating to IHC. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of David Haagsma Sent: Monday, February 06, 2006 7:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Immunofluorescence Text Dear Histonet; We have a customer that is looking for a good immuno book. We currently do not offer a book of this type and I do not have any recommendations for her - can you help her out? Thank you, Dave Haagsma MT(ASCP) MarketLab Inc. Subject: Find Product Form name...........: Caroline Ghaleb suggestion.....: Dear Sir, You have a good number of educational book, unfortunately I'm looking to buy a book about immunofluorescence. I appreciate if you can let me know if you can provide such a book. Thanks a lot. Caroline Ghaleb phone..........: email..........: teknolab@rcn.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Mon Feb 6 11:51:11 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Mon Feb 6 11:51:02 2006 Subject: [Histonet] Carstair's Stain Kits Message-ID: <83AACDB0810528418AA106F9AE9B7F7E013056C8@sjhaexc02.sjha.org> Does anyone know of a source for Carstair's in a kit? Thanks in advance. j Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From Ronnie_Houston <@t> bshsi.com Mon Feb 6 11:56:30 2006 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Mon Feb 6 12:00:28 2006 Subject: [Histonet] Immunofluorescence Text Message-ID: <00FBB1F3374BE24AAB7ACC6FCC7C617F561F4C@bsrexms01.bshsir.com> the best text I have come across dealing with Immunofluorescence is: Protein Localization by Fluorescence Microscopy: a practical approach ed. VJ Allan Oxford University Press 2000 ISBN 0-19-963741-5 (hbk) 0-19-963740-7 (pbk) Ronnie Houston Director of Anatomic Pathology Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 2877972 (804) 2877906 - fax ronnie_houston@bshsi.com -----Original Message----- From: Patsy Ruegg [mailto:pruegg@ihctech.net] Sent: Monday, February 06, 2006 12:51 PM To: 'David Haagsma'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Immunofluorescence Text The NSH IHC Resource Group has put together a list of reference books for IHC. I don't think there is specifically one on just F-IHC but several of them relate. You can join the IHCRG online at www.ihcrg.org for this and many other benefits relating to IHC. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of David Haagsma Sent: Monday, February 06, 2006 7:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Immunofluorescence Text Dear Histonet; We have a customer that is looking for a good immuno book. We currently do not offer a book of this type and I do not have any recommendations for her - can you help her out? Thank you, Dave Haagsma MT(ASCP) MarketLab Inc. Subject: Find Product Form name...........: Caroline Ghaleb suggestion.....: Dear Sir, You have a good number of educational book, unfortunately I'm looking to buy a book about immunofluorescence. I appreciate if you can let me know if you can provide such a book. Thanks a lot. Caroline Ghaleb phone..........: email..........: teknolab@rcn.com _______________________________________________ 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 ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone, and permanently delete the original e-mail, attachment(s), and any copies. From mlb <@t> nmr.mgh.harvard.edu Mon Feb 6 12:47:25 2006 From: mlb <@t> nmr.mgh.harvard.edu (mlb@nmr.mgh.harvard.edu) Date: Mon Feb 6 12:47:37 2006 Subject: [Histonet] "MBS" in Luxol Fast Blue MBS Message-ID: <1146.132.183.203.6.1139251645.squirrel@mail.nmr.mgh.harvard.edu> Hi, I had tried sending this message upon joining histonet, but I didn't see it in the archive. That could be why I haven't gotten a reply yet! Here it is again: Hi, I've been using luxol fast blue MBS for myelin staining and would like to know what the acronym "MBS" represents. I have read many primary sources, such as articles by Kluver and Barrera as well as Salthouse, and I have performed internet searches with Google, all to no avail (other than finding histonet!). Thanks for your help! Megan =-) p.s. Also, any explanations of the other acronyms used in luxol dyes (ARN, G and the "N" in MBSN) would be much appreciated! From gcallis <@t> montana.edu Mon Feb 6 12:59:27 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Feb 6 12:59:36 2006 Subject: [Histonet] Poster on RE: Color coding In-Reply-To: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDF82@EMAIL.archildrens .org> References: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDF82@EMAIL.archildrens.org> Message-ID: <6.0.0.22.1.20060206115815.01b4cab0@gemini.msu.montana.edu> Hazel, I remember your excellent poster, it was very informative. Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From SBarnes <@t> elch.org Mon Feb 6 13:34:53 2006 From: SBarnes <@t> elch.org (Sue Barnes) Date: Mon Feb 6 13:35:02 2006 Subject: [Histonet] NEW TISSUE PROCESSOR Message-ID: We are in the market for a new tissue processor. I am looking for one that is compatable with recycling of alcohol. Could I please hear from those of you who recycle alcohol and how well your processor works for you. Susan Barnes East Liverpool City Hospital East Liverpool, Ohio From jqb7 <@t> cdc.gov Mon Feb 6 13:42:53 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Mon Feb 6 13:52:40 2006 Subject: [Histonet] "MBS" in Luxol Fast Blue MBS Message-ID: Luxol fast blue MBSN is the chemical description for the specific dye, Solvent Blue 38 C32H12CuN8Na2O6S2 F.W. 732.16 Jeanine Bartlett, BS, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of mlb@nmr.mgh.harvard.edu Sent: Monday, February 06, 2006 1:47 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] "MBS" in Luxol Fast Blue MBS Hi, I had tried sending this message upon joining histonet, but I didn't see it in the archive. That could be why I haven't gotten a reply yet! Here it is again: Hi, I've been using luxol fast blue MBS for myelin staining and would like to know what the acronym "MBS" represents. I have read many primary sources, such as articles by Kluver and Barrera as well as Salthouse, and I have performed internet searches with Google, all to no avail (other than finding histonet!). Thanks for your help! Megan =-) p.s. Also, any explanations of the other acronyms used in luxol dyes (ARN, G and the "N" in MBSN) would be much appreciated! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sjchtascp <@t> yahoo.com Mon Feb 6 14:04:07 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Mon Feb 6 14:04:16 2006 Subject: [Histonet] small dryer Message-ID: <20060206200407.41079.qmail@web90210.mail.scd.yahoo.com> Does anyone know where I might be able to get a small forced air dryer, holding 2-3 racks of slides? Steve --------------------------------- Brings words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. From mariac <@t> creighton.edu Mon Feb 6 15:01:37 2006 From: mariac <@t> creighton.edu (Maria Christensen) Date: Mon Feb 6 15:07:58 2006 Subject: [Histonet] small dryer In-Reply-To: <20060206200407.41079.qmail@web90210.mail.scd.yahoo.com> References: <20060206200407.41079.qmail@web90210.mail.scd.yahoo.com> Message-ID: >Does anyone know where I might be able to get a >small forced air dryer, holding 2-3 racks of >slides? > > Steve > > >--------------------------------- >Brings words and photos together (easily) with > PhotoMail - it's free and works with Yahoo! Mail. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet We have one from TBS, Slide Dryer II. It works pretty well for what we use it (dry 130-160 slides O.N. at 37C) but it can dry slides in about half an hour. I think it lists for $1500 at Fisher Scientific, but we get a nice discount from them. Hope this helps! -- Mar?a Maria A. Christensen Technical Associate Dept. of Medical Microbiology & Immunology Creighton University School of Medicine 2500 California Plaza Omaha, Nebraska 68178-0404 voice (402) 280-2678 e-mail mariac@creighton.edu From portera <@t> msu.edu Mon Feb 6 13:25:02 2006 From: portera <@t> msu.edu (Amy Porter) Date: Mon Feb 6 15:35:48 2006 Subject: [Histonet] Immunohistochemistry on mouse teeth References: <819189266B78D543A02D9D3956EE4F6C030CEC99@addax.win.uthscsa.edu> Message-ID: <001001c62b53$08344100$8e7a0923@HistoJJ> Diane - Are you using a mouse primary? Didn't make a comment about this in your original post. If you are you would want to be using mouse on mouse staining techniques or kits. We always use another piece of tissue from the same animal if possible as a postive control, usually something from G.I. tract. Lots of proliferation going on there. Hope this helps out, ----- Original Message ----- From: "Horn, Diane A" To: Sent: Sunday, February 05, 2006 11:19 PM Subject: [Histonet] Immunohistochemistry on mouse teeth I am currently doing IHC on develping mouse teeth and I am getting only backgroung staining. The tissues are fixed O/N in 4% paraformaldehyde @4 degrees and embedded in paraffin. I have cut my sections @ 4um. I have tried HIER with Vector unmasking solution (boiling) for 2 minutes and using a kit for PCNA from Zymed. Has anyone done IHC on teeth that could give me some advice on what to try next? Thanks! Diane _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From barry_m <@t> ozemail.com.au Mon Feb 6 15:51:24 2006 From: barry_m <@t> ozemail.com.au (Barry Madigan) Date: Mon Feb 6 15:51:45 2006 Subject: [Histonet] MSH6 In-Reply-To: Message-ID: <000001c62b67$7a6eb7a0$0201010a@WORKSTATION1> Hello Josie, The antibodies for mismatch repair genes that we use consists of MLH1, MSH2 and MSH6 all of which we obtain from BD (Becton Dickinson). They are all performed on our BOND MAX Immunostainer using the high pH retrieval solution with EDTA (30 min) and a Vision Polymer Detection System. We have no troubles with them except sometimes we find that with archival cases we need to apply extra heat for MLH1. When this extra step is performed the MLH1 internal controls come up beautifully. Before we had the BOND MAX we used to heat retrieve using a microwave pressure cooker for 8 minutes on high in TRIS/EDTA pH 9.0 for MLH1. For the MSH6 antibody we currently obtain a dilution of 1:200. MSH2 1:500 and MLH1 1:100. Hope this is of some help. Regards Barry Barry Madigan Immunohistochemistry QHPS-Royal Brisbane Hospital Campus Australia -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Orr, Rebecca Sent: Saturday, 4 February 2006 1:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] MSH6 Hi Josie, How many different types of protocols did you try on the XT? With the different temp and Cell conditioning options I'm surprised you didn't get something. Are you running the MLH-1 and MSH-2 as well? How are they? My experience with the Microsateliite Instabilities is 4+ staining in 2 out of 3 antibodies, depending on the control tissue. The third antibody would compare at 2+ or a bit stronger. I have the best of both worlds with a Benchmark and a Nemesis. I experienced the poor staining results of these particular antibodies and I decided to run them on the Nemesis with Biocare polymer detection and they are gorgeous. I use the Biocare antibodies, too. In case you don't have that option, I would consider trying another control tissue, and running through all the CC options. I'm not sure if they behave differently without heat, did you try running a protocol with out heat in the detection? Do you have access to the polymer from Ventana? Maybe try that? Or run the antibody at 1:5 or 1:10. Or do you have a pressure cooker to try the HEIR offline. If you get good staining with offline HEIR then you can rule out the detection as the issue, too. Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 -----Original Message----- Message: 8 Date: Wed, 1 Feb 2006 13:54:36 -0600 From: "Nava, Josefa" Subject: [Histonet] looking for Antibodies-MSH6 and PMS2 To: Message-ID: <2C515C1049EAF5459EFD8C9B929078A41945A8@phdex03.txhealth.org> Content-Type: text/plain; charset="us-ascii" Hello Everyone , I am using Ventana BMK/XT , can someone tell me a good source of MSH6 and PMS2 that will work on the Ventana Machine. I appreciate any information. Thank you. Josie _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Mon Feb 6 16:05:30 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Mon Feb 6 16:05:39 2006 Subject: [Histonet] Immunohistochemistry on mouse teeth In-Reply-To: <001001c62b53$08344100$8e7a0923@HistoJJ> Message-ID: <200602062205.k16M5NDZ009781@chip.viawest.net> Have you decalcified the mouse teeth (I would imagine that you did)? How was the performed? Decal can have a determental effect on IHC. I use formic acid for good IHC results. If the calcium is not removed it can block access to the antigen site. Just some thoughts. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Porter Sent: Monday, February 06, 2006 12:25 PM To: Horn, Diane A; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Immunohistochemistry on mouse teeth Diane - Are you using a mouse primary? Didn't make a comment about this in your original post. If you are you would want to be using mouse on mouse staining techniques or kits. We always use another piece of tissue from the same animal if possible as a postive control, usually something from G.I. tract. Lots of proliferation going on there. Hope this helps out, ----- Original Message ----- From: "Horn, Diane A" To: Sent: Sunday, February 05, 2006 11:19 PM Subject: [Histonet] Immunohistochemistry on mouse teeth I am currently doing IHC on develping mouse teeth and I am getting only backgroung staining. The tissues are fixed O/N in 4% paraformaldehyde @4 degrees and embedded in paraffin. I have cut my sections @ 4um. I have tried HIER with Vector unmasking solution (boiling) for 2 minutes and using a kit for PCNA from Zymed. Has anyone done IHC on teeth that could give me some advice on what to try next? Thanks! Diane _______________________________________________ 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 From jkiernan <@t> uwo.ca Mon Feb 6 16:12:53 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Mon Feb 6 16:13:04 2006 Subject: [Histonet] "MBS" in Luxol Fast Blue MBS References: <1146.132.183.203.6.1139251645.squirrel@mail.nmr.mgh.harvard.edu> Message-ID: <43E7C9E5.B2FC7F08@uwo.ca> Your message appeared all right, but nobody answered. It probably means nobody knows what the MBS stands for. Luxol fast blues are not a chemical group of dyes. They are arylguanidinium salts of anionic dyes (rather than the usual sodium salts). This makes the dyes soluble in alcohol but not in water. When sections are stained, the organic cation stays in the alcohol, and the coloured anion lodges in the rather myelin. Like any other anionic dye, a luxol can be differentiated by alkali. Luxol fast blue MBS is a phthalocyanine dye; luxol fast blues G and ARN are azo dyes. See Clasen et al. 1973 J. Neuropath. Exp. Neurol. 32:271-283 for more on this subject, including how to make your own luxol-type dyes. John Kiernan London, Canada ----------------------------- mlb@nmr.mgh.harvard.edu wrote: > > Hi, I had tried sending this message upon joining > histonet, but I didn't see it in the archive. That > could be why I haven't gotten a reply yet! Here it > is again: > > Hi, I've been using luxol fast blue MBS for myelin > staining and would like to know what the acronym "MBS" > represents. > > I have read many primary sources, such as articles by > Kluver and Barrera as well as Salthouse, and I have > performed internet searches with Google, all to no > avail (other than finding histonet!). > > Thanks for your help! > Megan > =-) > > p.s. Also, any explanations of the other acronyms used > in luxol dyes (ARN, G and the "N" in MBSN) would > be much appreciated! > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From becky.garrison <@t> jax.ufl.edu Mon Feb 6 16:14:00 2006 From: becky.garrison <@t> jax.ufl.edu (Garrison, Becky) Date: Mon Feb 6 16:14:33 2006 Subject: [Histonet] CPT codes for muscle biopsies Message-ID: Could any of you share the CPT codes used for muscle biopsy work ups? My question is in 2 parts: 1) We currently charge: 88314 per stain for ATP, NADH, Esterase, Alkaline Phosphatase, Acid Phosphatase, AMPDA, Myophorylase, Cyto C Oxidase, and SDH on snap frozen muscle tissue and 88313 per stain for Trichrome, PAS, Oil Red O (ORO) on snap frozen muscle tissue in addition to 88305 for the biopsy itself. One pathologist is suggesting that we should be charging 88319 for the ATP thru SDH above and 88314 for the Trichrome thru ORO above and that 88313 to be used only on Trichromes, PAS and ORO on paraffin embedded tissue (such as for the nerve biopsies). In reviewing the 2006 CPT code book, it does seem that we may be charging incorrectly. 2) If 88319 is the correct answer to the first part: 88319 is not listed as an add on code. Do you still charge an 88305 for the biopsy? How would you charge a muscle biospsy, received fresh, divided into 2 parts as follows: First part is snap frozen in isopentane; frozen sections are stained with ATP x 3, SDH, Trichrome, ORO. Second part is submitted for routine processing and paraffin sections are stained with H&E, Trichrome and ORO? Thanks for your input. Becky Garrison Pathology Supervisor Jacksonville, FL 904-244-6237 From tkngflght <@t> yahoo.com Mon Feb 6 18:01:57 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Mon Feb 6 18:02:05 2006 Subject: [Histonet] Can anyone explain or at least empathise? In-Reply-To: <43E7C9E5.B2FC7F08@uwo.ca> Message-ID: <20060207000157.95181.qmail@web50913.mail.yahoo.com> Hey Fellow Histonetters: I read all the postings on here and generally understand all of it, but the more I read the more I realize I've forgotten more than I probably remember. (I haven't done a muscle stain group in quite a while!!). Does anyone else feel this way??? (Yes, I've passed 40, I'm sure someone out there can explain the mechanism of memory loss for me!!!) Thanks for slowing down the brain drain!! Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one tech at a time. admin@fullstaff.org 281.852.9457 office 281.883.7704 cell From tkngflght <@t> yahoo.com Mon Feb 6 18:01:57 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Mon Feb 6 18:02:07 2006 Subject: [Histonet] Can anyone explain or at least empathise? In-Reply-To: <43E7C9E5.B2FC7F08@uwo.ca> Message-ID: <20060207000157.95181.qmail@web50913.mail.yahoo.com> Hey Fellow Histonetters: I read all the postings on here and generally understand all of it, but the more I read the more I realize I've forgotten more than I probably remember. (I haven't done a muscle stain group in quite a while!!). Does anyone else feel this way??? (Yes, I've passed 40, I'm sure someone out there can explain the mechanism of memory loss for me!!!) Thanks for slowing down the brain drain!! Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one tech at a time. admin@fullstaff.org 281.852.9457 office 281.883.7704 cell From gangli.phd <@t> gmail.com Mon Feb 6 18:36:09 2006 From: gangli.phd <@t> gmail.com (Gang Li) Date: Mon Feb 6 18:36:17 2006 Subject: [Histonet] Barcode Labeling on Tissue Slides Message-ID: <120b7bf0602061636l18f78fb7m315a28e5c321e5c6@mail.gmail.com> Could anybody kindly tell me the barcode standards used for tissue slides labeling? Best wishes Gang Li, Ph.D. Biomedical Photometrics Inc. Waterloo, Ontario, CANADA From brucea <@t> unimelb.edu.au Mon Feb 6 20:14:52 2006 From: brucea <@t> unimelb.edu.au (Bruce Abaloz) Date: Mon Feb 6 20:15:19 2006 Subject: [Histonet] Kangaroo Mammary Tissue Message-ID: Dear Histonetter's, I am looking for stains that will identify the basic contents of cultured, mammary alveoli (mammospheres) of Kangaroo. Broad lipid ( ANY FAT STAIN THAT CAN BE DONE ON PARAFFIN EMBEDDED TISSUE???POSSIBLE??), carbohydrate (PAS?? ANY OTHER SUGGESTIONS) and protein stains (???) are required to determine what - IF any, different hormone combinations have/ effect on milk production. Any suggestions would be wonderful......our Histologist has made several good suggestions for me to try & we are both interested to see 'WHAT' POSSIBILITIES/suggestions the scientific minds in Histo-land come up with. Thankyou in advance & Cheers, Sonia -- BRUCE ABALOZ PH:61383446282 HISTOLOGIST FAX:61383447909 DEPT.of ZOOLOGY EMAIL: brucea@unimelb.edu.au THE UNIVERSITY Of MELBOURNE. VICTORIA. AUSTRALIA 3010 Nobody Can Make You Feel Inferior Without YOUR Permission - Eleanor Roosevelt DANCE LIKE NO-ONE'S WATCHING Q: What's a specimen? A: An Italian astronaut. ******************************************************************************** This email and any attachments may contain personal information or information that is otherwise privileged, confidential or otherwise protected from disclosure/subject of copyright. Any use, disclosure or copying of any part of it is prohibited. The University does not warrant that this email or any attachments are free from viruses or defects. Please check any attachments for viruses and defects before opening them. If this email is received in error please delete it and notify us by return email. The University does not necessarily share or endorse the views expressed in this email. From vanann702 <@t> skmc.gov.ae Mon Feb 6 22:11:12 2006 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Mon Feb 6 22:10:33 2006 Subject: [Histonet] NEW TISSUE PROCESSOR Message-ID: <7E070A7B959A9F42BE732545E5CF6210948D4D@SKMCEMAIL.skmc.gov.ae> We use recycled alcohol and have a Sakura VIP5 - in my opinion they are the best - no problems whatsoever. Recycled alcohol should not make any difference. Annieinarabia -----Original Message----- From: Sue Barnes [mailto:SBarnes@elch.org] Sent: Monday, February 06, 2006 11:35 PM To: Histonet (E-mail) Subject: [Histonet] NEW TISSUE PROCESSOR We are in the market for a new tissue processor. I am looking for one that is compatable with recycling of alcohol. Could I please hear from those of you who recycle alcohol and how well your processor works for you. Susan Barnes East Liverpool City Hospital East Liverpool, Ohio _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo.rogerson <@t> waht.swest.nhs.uk Tue Feb 7 02:48:32 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Tue Feb 7 02:48:44 2006 Subject: [Histonet] Can anyone explain or at least empathise? Message-ID: I've rationalised from the following standpoint; When you are younger you usually have less to think about and therefore you can focus your thoughts better. As you get older you have more to think about and usually less time and your thoughts become unfocused. I mean my son arranged perfectly easily to organise his snow boarding holiday but fails regularly to pay his rent. I think when you get older you also care more about forgetting; damn where are my glasses!!! Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Cheryl [mailto:tkngflght@yahoo.com] Sent: Tuesday, February 07, 2006 12:02 AM To: histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: [Histonet] Can anyone explain or at least empathise? Hey Fellow Histonetters: I read all the postings on here and generally understand all of it, but the more I read the more I realize I've forgotten more than I probably remember. (I haven't done a muscle stain group in quite a while!!). Does anyone else feel this way??? (Yes, I've passed 40, I'm sure someone out there can explain the mechanism of memory loss for me!!!) Thanks for slowing down the brain drain!! Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one tech at a time. admin@fullstaff.org 281.852.9457 office 281.883.7704 cell _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jorge.tornero <@t> gmail.com Tue Feb 7 06:32:52 2006 From: jorge.tornero <@t> gmail.com (Jorge Tornero) Date: Tue Feb 7 06:33:02 2006 Subject: [Histonet] Technovit 7100 resin removal Message-ID: <8c964a790602070432r457655a2x@mail.gmail.com> Hi all, I would like to remove the resin in my slides prior to stain. Do you know any protocol to do this? Thank you very much in advance Jorge Tornero IEO C?diz - Spain From jhaviland <@t> mdanderson.org Tue Feb 7 08:43:35 2006 From: jhaviland <@t> mdanderson.org (jhaviland@mdanderson.org) Date: Tue Feb 7 08:43:43 2006 Subject: [Histonet] troubles with CDK4 Message-ID: Hello: I am trying to do IHC with rabbit anti human CDK4 (D-22) from Santa Cruz on salivary gland tissue. I am having background problems which is leading to difficulty in figuring out what is positive and what is negative. I have used LSAB2 kit from Dako, Rabbit Envision Polymer from Dako and the Rabbit ABC Elite kit from Vector. I do a 3% hydrogen peroxide treatment for 20 minutes and do antigen retrieval with 10mM Citric Buffer pH 6.0 for 45 minutes in a steamer and a 20 minute cool down. I do a blocking step and have used either serum free or goat serum for a block. I already checked with the company about my protocol and it is the same as theirs. I tried doing it on skin which is reccommended as a control but still have background problems. Thanks Joie Haviland Senior Research Assistant MD Anderson Cancer Research Center From Susan.Ferrigon <@t> sanofi-aventis.com Tue Feb 7 08:53:48 2006 From: Susan.Ferrigon <@t> sanofi-aventis.com (Susan.Ferrigon@sanofi-aventis.com) Date: Tue Feb 7 08:54:05 2006 Subject: [Histonet] Retic staining on rodent livers Message-ID: Does anyone have problems with staining for reticulin on rodent livers?? if not, do you have a particular method you use?? I use Gordon and Sweet and find the fibres don't stain up very well and the background is grainy. Any advise. Thanks Susan ------------------------------ Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de ce courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuellement pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ------------------------------ From p.simpson <@t> improvision.com Tue Feb 7 09:04:07 2006 From: p.simpson <@t> improvision.com (Pippa Simpson) Date: Tue Feb 7 09:04:48 2006 Subject: [Histonet] Job posting : Sales and Support opportunities at Improvision Inc Message-ID: <017d01c62bf7$bedc6870$0701a8c0@improvision.com> Please forgive the commercial intrusion. Improvision is currently considering applicants for roles within our Sales and Technical Support teams. For further details please visit http://www.improvision.com/careers/ or contact recruitment@improvision.com. Please do not reply to the listserver. Kind regards Pippa Simpson Improvision Human Resources From julien_lambreydesouza <@t> uqar.qc.ca Tue Feb 7 09:50:25 2006 From: julien_lambreydesouza <@t> uqar.qc.ca (Julien Lambrey de Souza) Date: Tue Feb 7 09:50:36 2006 Subject: [Histonet] decalcifying before HBQ stain Message-ID: <75c6cee121c47a552bf01aa9b6910f1b@uqar.qc.ca> Hello everyone, Just a quick question: Is it necessary to decalcify before staining with the HBQ protocol? Thanks, Julien Lambrey de Souza Research assistant, Evolutionary biology University of Quebec at Rimouski Tel: (418) 723-1986 #1714 Fax: (418) 724-1849 From cpomajzl <@t> cpllabs.com Tue Feb 7 10:12:11 2006 From: cpomajzl <@t> cpllabs.com (Chris Pomajzl) Date: Tue Feb 7 10:10:54 2006 Subject: [Histonet] JOB OPENING IN AUSTIN TEXAS Message-ID: <006901c62c01$43393590$26fca8c0@CSP> Greetings: We have multiple shift openings in Austin, Texas. If you might be interested in a fast-paced, high-volume laboratory, please give me a call. Duties include embedding, sectioning, special stains, etc. Competetive salary, health benefits, and a friendly working environment. Thanks. Sincerely, Chris Pomajzl, HTL (ASCP) Histology Supervisor Clinical Pathology Laboratories, Inc. 512.873.1660 (o) 512.873.5004 (f) cpomajzl@cpllabs.com CONFIDENTIALITY NOTICE: This communication is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged, confidential, and exempt from disclosure under applicable law. If you are not the intended recipient, you are notified that any use, dissemination, distribution, or copying of this communication is strictly prohibited. If you have received this message in error, please notify the sender immediately and destroy this communication. From rkrug <@t> sial.com Tue Feb 7 10:17:43 2006 From: rkrug <@t> sial.com (Robert Krug) Date: Tue Feb 7 10:18:36 2006 Subject: [Histonet] Luxol dyes Message-ID: Megan: Saw Dr Kiernan's posting on Histonet. I cannot tell you what the MBS stands for, but I can add a little insight into naming dyes. Dyes are given trivial names in most cases. This really means the person originating the dye may call the dye whatever they please. At one time there were may have existed guidelines. If such guidelines existed, no one seems to be familiar with the specifics at this late date. Today we use scientific nomenclature with set conventions to name products such as 1-Cyclohexyl-2-phyrrolidone. Personally I prefer the trivial names. In some cases the abbreviations come from the German dye industry - who were really pioneers in the field. The SF in Light Green SF yellowish = SaureFarbstoff (German) = acid dye English terms are also sometimes abbreviated FCF in Fast Green FCF = For Coloring Food There are a series of fluorescent dyes available such as PKH26, PKH67. In this particular case I cannot be 100% sure, but the person who filed the patent on these dyes just happened to have the initials PKH. Coincidence? In this particular case I would guess that the 26th and the 67th attempts were keepers. Naphthol compounds are often named Naphthol AS-MX phosphate or possibly Naphthol AS-BI phosphate. The structures are evidently different, but no one I have talked to has been able to explain why one product is given as the AS-MX designation and the next product is AS-BI If you look at Conn's Biological Stains, 9th Edition, you will find the text lists many more obscure synonyms - as compared to the 10th Edition. In previous years, companies often gave common dyes unique names. They may have believed they gained some competitive advantage from this practice. Even today dyes may often go by more than one name. Sometime this is done when the company markets to various industries. If you are selling to the printing industry, you might name a dye XXXX. If you market dyes for histological use, you might name a dye YYYY. Today I believe the trend is for companies to sell the dye by the name given the most common product of commerce. This is probably why fewer synonyms are listed in the 10th Edition. Synonyms however are here to stay. Sodium thiosulfate is still referred to as hypo in older textbooks. This is because "hypo" was the term used in photography. One of the many useful functions served by the Biological Stain Commission was the creation of CI or Color Index numbers. It really doesn't matter what a company names a dye. If Product X and Product Y from competing companies have the same CI number, there is a good chance the dyes may be used in the same staining procedures. If the dyes are Certified by the BSC, this gives even more indication the dye should be acceptable for use. The CI number varies from the CAS number. For the CAS numbers to be identical, the molecular structure should be identical. Not so with Color Index numbers. For example, Basic Fuchsin may be composed of 100% Pararosaniline, or a mixture of pararosaniline, rosaniline, magenta II and magenta III. The pararosaniline may be the chloride or the acetate form. So although the blends may vary from manufacturer to manufacturer and possibly even between lot numbers for a specific manufacturer, the dye must perform in an acceptable manner to receive certification from the Biological Stain Commission. In the 10th Edition of Conn's, no CI number is assigned to Basic Fuchsin, although the individual homologs are given given unique homologs. The 9th Edition of Conn's assigned Basic Fuchsin the CI number for Pararosanline. The real test is how well the dyes perform in actual staining procedures. Anyone really interested in dyes for biological use should try to obtain a copy of the methods used by the Biological Stain Commission. Analysis and testing of biological stains - The Biological Stain Commission Procedures (Biotechnic & Histochemistry 2002, 77(5&6): 237-275 Methylene Blue is another prime example for a dye which is composed of a series of closely related homologs. However in this case Methylene Blue is assigned a Color Index number, as opposed to Basic Fuchsin which was not assigned a unique CI number in the 10th Edition of Conn's. Again the major homologs are assigned CI numbers. Other dyes by comparison are relatively pure and may be almost 100% pure. However sometimes purity is not all. Some of the impurities may be present/added/act as stabilizers. In the case of Nile Blue, the presence of Nile Red as a contaminant may be highly desirable. For other dyes the formulations are constantly shifting, even though the name remains relatively constant. Alcian Blue is a dye which has a tendency to explode during manufacture. This has resulted in a constant tinkering with the method of production over the years. The dye sold today was probably manufactured very differently than an Alcian Blue manufactured decades ago. If the product of commerce continues to provide acceptable staining results in the methods commonly associated with Alcian Blue, the dye will be sold & certified as Alcian Blue. It would be impractical to give each slight variation in product a unique name or CAS number. Trying to perform a literature search would be made extremely difficult to almost impossible. Tradition methyl green has not been produced commercially for many years (35-45+ ?). The actual product of commerce today is ethyl green. In this case methyl green and ethyl green are really not synonyms. However because ethyl green may be used for the same purposes as methyl green, the product of commerce continues to be called methyl green - very much to the annoyance of Dr Kiernan :) Hope this helps, Best Regards, Bob Krug Sigma-Aldrich St Louis, MO From bhewlett <@t> cogeco.ca Tue Feb 7 10:37:48 2006 From: bhewlett <@t> cogeco.ca (bhewlett@cogeco.ca) Date: Tue Feb 7 10:37:57 2006 Subject: [Histonet] decalcifying before HBQ stain Message-ID: <43e8ccdc.319.3f3d.9906@cogeco.ca> > Julien, Just what is the HBQ protocol? Bryan > Hello everyone, > > Just a quick question: Is it necessary to decalcify before staining > with the HBQ protocol? > > Thanks, > > > Julien Lambrey de Souza > Research assistant, Evolutionary biology > University of Quebec at Rimouski > > Tel: (418) 723-1986 #1714 > Fax: (418) 724-1849 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From julien_lambreydesouza <@t> uqar.qc.ca Tue Feb 7 11:50:31 2006 From: julien_lambreydesouza <@t> uqar.qc.ca (Julien Lambrey de Souza) Date: Tue Feb 7 11:50:41 2006 Subject: [Histonet] RE: Decalcifying before HBQ Message-ID: <634240d185312d7b6108d64157d0b1e0@uqar.qc.ca> Sorry. I guess I should have elaborated a bit on the stain. HBQ = Hall and Brunt's Quadruple stain Used to differenciate bone from cartillage and chondroid bone. Deparaffinized sections are stained in aqueous celestine blue (0,5% in 5% ammonium sulphate and 14% glycerin), Mayer's hematox, alcian blue (1% in 1% acetic acid, pH 2,6), Phosphomolybdic acid1% and aqueous direct red 0,5%. All the protocols and pictures I have seen in publications are done on decalcified tissue. My question is the following: If my specimen cuts out nice sections without decalcification, will I get the intended staining (same as seen in publications) or will it be different from decalcified sections? How does decalcification influence staining results? Thanks for the imput. Julien Lambrey de Souza Research assistant, Evolutionary biology University of Quebec at Rimouski Tel: (418) 723-1986 #1714 Fax: (418) 724-1849 From germckeon <@t> excite.com Tue Feb 7 11:52:02 2006 From: germckeon <@t> excite.com (germckeon@excite.com) Date: Tue Feb 7 11:52:13 2006 Subject: [Histonet] Technovit 7100 resin removal Message-ID: <20060207175202.C3E6137628@xprdmailfe14.nwk.excite.com> Hello Jorge, I am also trying to find out the same thing. Would it be possible to send me any hints you may get? Many thanks, Gerald McKeon Trinity College, Dublin, Ireland. [Histonet] Technovit 7100 resin removal Hi all, I would like to remove the resin in my slides prior to stain. Do you know any protocol to do this? Thank you very much in advance Jorge Tornero IEO Cαdiz - Spain _______________________________________________ Join Excite! - http://www.excite.com The most personalized portal on the Web! From liz <@t> premierlab.com Tue Feb 7 12:21:05 2006 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Tue Feb 7 12:21:16 2006 Subject: [Histonet] prions and frozen sections Message-ID: <000001c62c13$4281a030$95d48a80@Chlipala> Is anyone familiar with the precautions that need to be taken when cutting frozen sections (unfixed, un treated) on prion infected tissue (CWD and scrapie) in deer and sheep, any advice would be helpful. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From TillRenee <@t> uams.edu Tue Feb 7 12:39:45 2006 From: TillRenee <@t> uams.edu (Till, Renee) Date: Tue Feb 7 12:40:28 2006 Subject: [Histonet] lysozyme Message-ID: <11F927674DEBDC43B960809A7403C5D232805E@MAILPED.ad.uams.edu> Does anyone know of a lysozyme antibody that will stain mouse tissue? Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 ================================================================================================ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ================================================================================================ From RJLevier <@t> LancasterGeneral.org Tue Feb 7 12:44:47 2006 From: RJLevier <@t> LancasterGeneral.org (LeVier, Rebecca J) Date: Tue Feb 7 12:45:02 2006 Subject: [Histonet] Cutting Message-ID: <0FDBF29200C637468CCC1F9E7E85A3D214E8FD@MAIL-LR.lha.org> Hello Histonetters....... I have a question regarding blocks and how many each person is cutting per day. Does anyone have a limit that they find to be acceptable and how many would you consider to be too many to cut in one day by one person? Also with that could any of you help me with how many techs you have and the amount of total blocks you have on an average day and do all of the techs cut or is there set responsibilities that only on some cut? Thank you in advance for all the help. Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From RJLevier <@t> LancasterGeneral.org Tue Feb 7 12:48:22 2006 From: RJLevier <@t> LancasterGeneral.org (LeVier, Rebecca J) Date: Tue Feb 7 12:48:41 2006 Subject: [Histonet] Cassette Labeling Message-ID: <0FDBF29200C637468CCC1F9E7E85A3D214E8FE@MAIL-LR.lha.org> Could anyone give me some feedback on the Leica Cassette Labeler or the Sakura Cassette Labeler? Thanks Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From dndsomi <@t> aol.com Tue Feb 7 12:49:38 2006 From: dndsomi <@t> aol.com (dndsomi@aol.com) Date: Tue Feb 7 12:49:53 2006 Subject: [Histonet] Morgue Cooler Temperature Message-ID: <8C7FA2E61A40630-AE4-4D4E@FWM-D20.sysops.aol.com> Can someone please give me a reference for CAP morgue cooler temperature ranges? Thanks From Barry.R.Rittman <@t> uth.tmc.edu Tue Feb 7 13:15:34 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Tue Feb 7 13:15:37 2006 Subject: [Histonet] lysozyme Message-ID: Renee Although I cannot suggest an antibody, have you considered using Lendrum's phloxine tartrazine for this? Barry From nancy.troiano <@t> yale.edu Tue Feb 7 13:33:22 2006 From: nancy.troiano <@t> yale.edu (Nancy W. Troiano) Date: Tue Feb 7 13:33:35 2006 Subject: [Histonet] Technovit 7100 resin removal Message-ID: <5.2.1.1.2.20060207143225.00c38b48@email.med.yale.edu> I don't know of any way to remove Technovit 7100 resin (GMA) prior to staining. We stain right through it. You'd have to embed in methylmethacrylate if you want a resin that is removable prior to staining. From TJJ <@t> Stowers-Institute.org Tue Feb 7 13:43:04 2006 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Tue Feb 7 13:43:26 2006 Subject: [Histonet] Re: troubles with CDK4 Message-ID: Dear Joie, I can't find a Cdk4 (D-22) antibody in their catalog. I see a C-22 and an H-22. If you are using the C-22, which they indicate can be used in IHC-paraffin embedded material, call them and ask what their staining conditions are (including fixative used). The detection reagents you are using might be ultra sensitive (too sensitive) for this antibody. Either extend the dilution of the primary antibody, or try using an HRP conjugated anti-rabbit secondary antibody at the concentration you are currently using to see if that lowers the background staining. The staining intensity in the picture in the Santa Cruz catalog of the C-22 on human skin is very weak. Perhaps that's the best signal they can get with this antibody while keeping the background noise in check. You might also try pre-blocking the primary antibody by adding 2-5% serum concentration of the species you are staining to the antibody diluent you are using. Make your antibody dilution in this and let it incubate in the tube prior to placing it on the slide. There are a variety of things you can try, but be aware that you may never get it working suitably despite the company's claims. You might need to look elsewhere for a different source. Gayle Callis turned me on to this site: http://www.exactantigen.com/ and another good one is http://www.abcam.com Good luck! Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From michael.owen <@t> fda.hhs.gov Tue Feb 7 14:20:08 2006 From: michael.owen <@t> fda.hhs.gov (Owen, Michael P) Date: Tue Feb 7 14:21:11 2006 Subject: [Histonet] prions and frozen sections Message-ID: From: Owen, Michael P Sent: Tuesday, February 07, 2006 10:27 AM To: ABSA: Biosafety Discussion List Subject: FW: [Histonet] prions and frozen sections Dear Biosafety List Members, Anyone want to take a shot at this question from Histonet? From: Sonia Rosenberger [mailto:srosen@berkeley.edu] Sent: Tuesday, February 07, 2006 12:08 PM To: liz@premierlab.com Cc: Owen, Michael P Subject: Re: FW: [Histonet] prions and frozen sections See the guidance "Best Management Practices for Handling TSE Tissues" for CWD tissues at http://www.aavld.org/aavld-3/current_news.jsp. Cheers, Sonia Michael P. Owen, Regulatory Microbiologist U.S. FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.hhs.gov From gcallis <@t> montana.edu Tue Feb 7 14:37:01 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Feb 7 14:37:26 2006 Subject: [Histonet] Technovit 7100 resin removal In-Reply-To: <20060207175202.C3E6137628@xprdmailfe14.nwk.excite.com> References: <20060207175202.C3E6137628@xprdmailfe14.nwk.excite.com> Message-ID: <6.0.0.22.1.20060207133426.01b3f750@gemini.msu.montana.edu> Techovits 7100 is a Glycol methacrylate (GMA) formulation and once polymerized, cannot be removed from the section or etched either. Water may soften the plastic but it does NOT dissolve it away. If you want to remove a plastic from a section for Immunostaining, then the Technovits 9100, methyl methacrylate would be a better choice for embedding, then use 60C xylene or a solvent that will dissolve the MMA away. At 10:52 AM 2/7/2006, you wrote: >Hello Jorge, > >I am also trying to find out the same thing. Would it be possible to send >me any hints you may get? > > > >Many thanks, > >Gerald McKeon > >Trinity College, > >Dublin, Ireland. > > > > > >[Histonet] Technovit 7100 resin removal > > > >Hi all, > > > >I would like to remove the resin in my slides prior to stain. Do you > >know any protocol to do this? > > > >Thank you very much in advance > > > >Jorge Tornero > >IEO C?diz - Spain > > > >_______________________________________________ >Join Excite! - http://www.excite.com >The most personalized portal on the Web! > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From pruegg <@t> ihctech.net Tue Feb 7 14:53:14 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Tue Feb 7 14:53:19 2006 Subject: [Histonet] Technovit 7100 resin removal In-Reply-To: <6.0.0.22.1.20060207133426.01b3f750@gemini.msu.montana.edu> Message-ID: <200602072053.k17Kr5DZ016078@chip.viawest.net> True Gayle, there are techniques to etch or soften the GMA but it cannot be dissolved. We used to use a very strong ethoxide (alcohol in sodium hydroxide) solution to etch before staining (this method does not fair well for IHC). As you mentioned, the best bet to do IHC is to use MMA. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Tuesday, February 07, 2006 1:37 PM To: germckeon@excite.com; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Technovit 7100 resin removal Techovits 7100 is a Glycol methacrylate (GMA) formulation and once polymerized, cannot be removed from the section or etched either. Water may soften the plastic but it does NOT dissolve it away. If you want to remove a plastic from a section for Immunostaining, then the Technovits 9100, methyl methacrylate would be a better choice for embedding, then use 60C xylene or a solvent that will dissolve the MMA away. At 10:52 AM 2/7/2006, you wrote: >Hello Jorge, > >I am also trying to find out the same thing. Would it be possible to >send me any hints you may get? > > > >Many thanks, > >Gerald McKeon > >Trinity College, > >Dublin, Ireland. > > > > > >[Histonet] Technovit 7100 resin removal > > > >Hi all, > > > >I would like to remove the resin in my slides prior to stain. Do you > >know any protocol to do this? > > > >Thank you very much in advance > > > >Jorge Tornero > >IEO C?diz - Spain > > > >_______________________________________________ >Join Excite! - http://www.excite.com >The most personalized portal on the Web! > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Feb 7 15:20:11 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Feb 7 15:20:28 2006 Subject: [Histonet] prions and frozen sections In-Reply-To: References: Message-ID: <6.0.0.22.1.20060207140121.01b6bb30@gemini.msu.montana.edu> Our laboratory is BSL2 for prion work, murine and hamster animal models - brain, spinal cord and other tissues. A microtomy room is dedicated to prion work, and also contains a cryostat dedicated to prion work ONLY whether the tissus is fixed or not prior to cryomicrotomy. There is no way we can totally, 100% decontaminate the cryostat for prions, BSL2 or not, and I have too many other people, including students and technicians working on another cryostat and have to worry about prion decontamination. The researcher bought and maintains his own cryostat and hopefully it has a long life as no repairman would go inside it now. For paraffin work (also cryostat), workers take the necessary precautions, gloves, safety glasses, disposable lab coats, etc. when sectioning paraffin blocks and frozen sections. The microtome, waterbath are also dedicated to prion, nothing else is cut on these. Water from waterbath is dumped into a carboy of 6N NaOH (sitting in a big tub to collect any spills) until it reaches a volume where the NaOH becomes 2N simply by dilution, all is allowed to sit. We used to use bleach, that has gone by the wayside in favor of the NaOH. Carboy is then picked up by our biohazard safety laboratory on campus. Dry waste, towels, wipes, etc are collected for incineration i.e paraffin trimmings also. If we worked with sheep or CWD, then we would put sticky floor mats in front of doorway, and under microtome area. Trimmings should stay in the room. Sounds like a very uptight attitude for handling prions, but better to take stricter, proper precautions now and then have to change later. Some people collect alcohols into bleach but when we did this, we noticed a huge heat reaction - that does NOT happen with NaOH so we opted for the latter just not not have the heat problem. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From jqb7 <@t> cdc.gov Tue Feb 7 15:18:34 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Tue Feb 7 15:32:01 2006 Subject: [Histonet] prions and frozen sections References: Message-ID: I have replied privately. In short, I feel unless you have a dedicated room and equipment this should not be handled in a routine lab. Jeanine Bartlett, BS, HT(ASCP) Centers for Disease Control Atlanta, GA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Owen, Michael P Sent: Tue 2/7/2006 3:20 PM To: 'Histonet' Cc: Subject: [Histonet] prions and frozen sections From: Owen, Michael P Sent: Tuesday, February 07, 2006 10:27 AM To: ABSA: Biosafety Discussion List Subject: FW: [Histonet] prions and frozen sections Dear Biosafety List Members, Anyone want to take a shot at this question from Histonet? From: Sonia Rosenberger [mailto:srosen@berkeley.edu] Sent: Tuesday, February 07, 2006 12:08 PM To: liz@premierlab.com Cc: Owen, Michael P Subject: Re: FW: [Histonet] prions and frozen sections See the guidance "Best Management Practices for Handling TSE Tissues" for CWD tissues at http://www.aavld.org/aavld-3/current_news.jsp. Cheers, Sonia Michael P. Owen, Regulatory Microbiologist U.S. FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.hhs.gov _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Tue Feb 7 15:39:04 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Tue Feb 7 15:39:09 2006 Subject: [Histonet] Technovit 7100 resin removal Message-ID: Sodium methoxide has been used to remove epoxy resin, will this not work on Technovit resin ? Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Tuesday, February 07, 2006 2:53 PM To: 'Gayle Callis'; germckeon@excite.com; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Technovit 7100 resin removal True Gayle, there are techniques to etch or soften the GMA but it cannot be dissolved. We used to use a very strong ethoxide (alcohol in sodium hydroxide) solution to etch before staining (this method does not fair well for IHC). As you mentioned, the best bet to do IHC is to use MMA. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Tuesday, February 07, 2006 1:37 PM To: germckeon@excite.com; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Technovit 7100 resin removal Techovits 7100 is a Glycol methacrylate (GMA) formulation and once polymerized, cannot be removed from the section or etched either. Water may soften the plastic but it does NOT dissolve it away. If you want to remove a plastic from a section for Immunostaining, then the Technovits 9100, methyl methacrylate would be a better choice for embedding, then use 60C xylene or a solvent that will dissolve the MMA away. At 10:52 AM 2/7/2006, you wrote: >Hello Jorge, > >I am also trying to find out the same thing. Would it be possible to >send me any hints you may get? > > > >Many thanks, > >Gerald McKeon > >Trinity College, > >Dublin, Ireland. > > > > > >[Histonet] Technovit 7100 resin removal > > > >Hi all, > > > >I would like to remove the resin in my slides prior to stain. Do you > >know any protocol to do this? > > > >Thank you very much in advance > > > >Jorge Tornero > >IEO C?diz - Spain > > > >_______________________________________________ >Join Excite! - http://www.excite.com >The most personalized portal on the Web! > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ 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 From pruegg <@t> ihctech.net Tue Feb 7 15:40:37 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Tue Feb 7 15:40:56 2006 Subject: [Histonet] troubles with CDK4 In-Reply-To: Message-ID: <200602072140.k17LegDZ028399@chip.viawest.net> Try using your serum free protein block before the secondary reagent as well as before the primary. In my experience most non specific staining is from one or more of 5 things: 1. The secondary or labeled polymer is binding non specifically to the tissue (this can be alleviated by using a really good f(ab)fragmented secondary which has been absorbed to the species of your primary antibody). 2. The endogenous peroxidase has not been quenched. 3. There is endogenous biotin left and you are using an AB detection system. 4. You have over treated by HIER or EIER (this often shows up as unexpected nuclear staining). 5. What is thought to be non-specific staining is actually specific binding to tissue IgG. I am sure there are many other causes for non specific staining, but in my experience these five are most common. Did you get the same BG with LSAB detection that you got with labeled polymer? If so, then it is not the endogenous biotin causing your problem. Also, to really find out, run your slides adding one step at a time to see where you get non specific staining. Forinstance, stain a set of slides leaving out the primary antibody for all, don't block with h202 and do the DAB to see what it looks like with out blocking, do the block for endogenous peroxidase and then go directly to DAB leaving out the secondary and primary steps and if you are still getting staining that means your peroxidase block is not working properly, keep adding the detection reagents without the primary antibody until you get staining. If you don't ever get staining until you add the primary antibody then you can assume that it is specific staining related to that antibody. Sometimes we get undesired "specific" staining, which we often call "non specific". Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of jhaviland@mdanderson.org Sent: Tuesday, February 07, 2006 7:44 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] troubles with CDK4 Hello: I am trying to do IHC with rabbit anti human CDK4 (D-22) from Santa Cruz on salivary gland tissue. I am having background problems which is leading to difficulty in figuring out what is positive and what is negative. I have used LSAB2 kit from Dako, Rabbit Envision Polymer from Dako and the Rabbit ABC Elite kit from Vector. I do a 3% hydrogen peroxide treatment for 20 minutes and do antigen retrieval with 10mM Citric Buffer pH 6.0 for 45 minutes in a steamer and a 20 minute cool down. I do a blocking step and have used either serum free or goat serum for a block. I already checked with the company about my protocol and it is the same as theirs. I tried doing it on skin which is reccommended as a control but still have background problems. Thanks Joie Haviland Senior Research Assistant MD Anderson Cancer Research Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pmarcum <@t> vet.upenn.edu Tue Feb 7 16:02:38 2006 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Tue Feb 7 16:02:55 2006 Subject: [Histonet] Technovit 7100 resin removal In-Reply-To: References: Message-ID: <6.1.1.1.2.20060207165758.019979d0@mail.vet.upenn.edu> Actually it will soften slightly and cause the GMA to curl and wrinkle badly however it will not remove it from the section. It may lift it off the slide. Unfortunately nothing has been found to successfully remove GMA and in fact it continues to polymerize as it ages making it even more difficult to remove each day. The polymerization even seems to continue on the slides once the sections are picked up. It has always seemed strange to me that Methoxide as we called it would remove epoxy and not GMA however it is a fact. Pam Marcum UPENN Vet School At 04:39 PM 2/7/2006, Rittman, Barry R wrote: >Sodium methoxide has been used to remove epoxy resin, will this not work >on Technovit resin ? >Barry > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg >Sent: Tuesday, February 07, 2006 2:53 PM >To: 'Gayle Callis'; germckeon@excite.com; Histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] Technovit 7100 resin removal > >True Gayle, there are techniques to etch or soften the GMA but it cannot be >dissolved. We used to use a very strong ethoxide (alcohol in sodium >hydroxide) solution to etch before staining (this method does not fair well >for IHC). As you mentioned, the best bet to do IHC is to use MMA. >Patsy > > >Patsy Ruegg, HT(ASCP)QIHC >IHCtech, LLC >Fitzsimmons BioScience Park >12635 Montview Blvd. Suite 216 >Aurora, CO 80010 >P-720-859-4060 >F-720-859-4110 >wk email pruegg@ihctech.net >web site www.ihctech.net > > >This email is confidential and intended solely for the use of the Person(s) >('the intended recipient') to whom it was addressed. Any views or opinions >presented are solely those of the author. It may contain information that is >privileged & confidential within the meaning of applicable law. Accordingly >any dissemination, distribution, copying, or other use of this message, or >any of its contents, by any person other than the intended recipient may >constitute a breach of civil or criminal law and is strictly prohibited. If >you are NOT the intended recipient please contact the sender and dispose of >this e-mail as soon as possible. > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis >Sent: Tuesday, February 07, 2006 1:37 PM >To: germckeon@excite.com; Histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Technovit 7100 resin removal > >Techovits 7100 is a Glycol methacrylate (GMA) formulation and once >polymerized, cannot be removed from the section or etched either. Water >may soften the plastic but it does NOT dissolve it away. If you want to >remove a plastic from a section for Immunostaining, then the Technovits >9100, methyl methacrylate would be a better choice for embedding, then use >60C xylene or a solvent that will dissolve the MMA away. > > At 10:52 AM 2/7/2006, you wrote: > > > > > > >Hello Jorge, > > > >I am also trying to find out the same thing. Would it be possible to > >send me any hints you may get? > > > > > > > >Many thanks, > > > >Gerald McKeon > > > >Trinity College, > > > >Dublin, Ireland. > > > > > > > > > > > >[Histonet] Technovit 7100 resin removal > > > > > > > >Hi all, > > > > > > > >I would like to remove the resin in my slides prior to stain. Do you > > > >know any protocol to do this? > > > > > > > >Thank you very much in advance > > > > > > > >Jorge Tornero > > > >IEO C?diz - Spain > > > > > > > >_______________________________________________ > >Join Excite! - http://www.excite.com > >The most personalized portal on the Web! > > > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Gayle Callis >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 >406 994-4303 (FAX) > > > >_______________________________________________ >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 Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From ander093 <@t> tc.umn.edu Tue Feb 7 16:13:49 2006 From: ander093 <@t> tc.umn.edu (LuAnn Anderson) Date: Tue Feb 7 16:14:11 2006 Subject: [Histonet] prions and frozen sections In-Reply-To: <6.0.0.22.1.20060207140121.01b6bb30@gemini.msu.montana.edu> References: <6.0.0.22.1.20060207140121.01b6bb30@gemini.msu.montana.edu> Message-ID: <6.2.3.4.0.20060207161245.01d9dc70@ander093.email.umn.edu> Ditto~~but absolutely no frozens allowed. LuAnn Anderson HT(ASCP) Neuropathology Lab University of Minnesota At 03:20 PM 2/7/2006, Gayle Callis wrote: >Our laboratory is BSL2 for prion work, murine and hamster animal >models - brain, spinal cord and other tissues. > >A microtomy room is dedicated to prion work, and also contains a >cryostat dedicated to prion work ONLY whether the tissus is fixed or >not prior to cryomicrotomy. There is no way we can totally, 100% >decontaminate the cryostat for prions, BSL2 or not, and I have too >many other people, including students and technicians working on >another cryostat and have to worry about prion >decontamination. The researcher bought and maintains his own >cryostat and hopefully it has a long life as no repairman would go >inside it now. > >For paraffin work (also cryostat), workers take the necessary >precautions, gloves, safety glasses, disposable lab coats, etc. when >sectioning paraffin blocks and frozen sections. The microtome, >waterbath are also dedicated to prion, nothing else is cut on these. > >Water from waterbath is dumped into a carboy of 6N NaOH (sitting in >a big tub to collect any spills) until it reaches a volume where the >NaOH becomes 2N simply by dilution, all is allowed to sit. We used >to use bleach, that has gone by the wayside in favor of the >NaOH. Carboy is then picked up by our biohazard safety laboratory >on campus. Dry waste, towels, wipes, etc are collected for >incineration i.e paraffin trimmings also. If we worked with sheep >or CWD, then we would put sticky floor mats in front of doorway, and >under microtome area. Trimmings should stay in the room. > >Sounds like a very uptight attitude for handling prions, but better >to take stricter, proper precautions now and then have to change later. > >Some people collect alcohols into bleach but when we did this, we >noticed a huge heat reaction - that does NOT happen with NaOH so we >opted for the latter just not not have the heat problem. > > > > >Gayle Callis >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 >406 994-4303 (FAX) > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From histolog <@t> fcv.unl.edu.ar Tue Feb 7 16:51:13 2006 From: histolog <@t> fcv.unl.edu.ar (Lab. Invest. Histol.) Date: Tue Feb 7 16:51:26 2006 Subject: [Histonet] PBS..??? In-Reply-To: <6.2.3.4.0.20060207161245.01d9dc70@ander093.email.umn.edu> Message-ID: I wanted to know that ventages and/or disadvantages have the different ones preparations of PBS for immunohistochemistry. The biggest questions are: Potassium phosphate monobasic or Sodium phosphate monobasic???? 0.1M, 0.02M or 0.01M ???? pH 7.00 , pH 7.20 or pH 7.40???? Which is the it formulates that you use? Thanks!!! Hugo ------------------------------------------------------------------- Dr. Hugo H. Ortega (DMV, PhD) Departament of Cellular Biology Faculty of Veterinary Sciences Universidad Nacional del Litoral R.P. Kreder 2805 - Esperanza (3080) Santa Fe - ARGENTINA Tel. (54)3496-420639 Fax. (54)3496-426304 http://fcv.unl.edu.ar/histolog/ http://fcv.unl.edu.ar/bioterio/ From julien_lambreydesouza <@t> uqar.qc.ca Tue Feb 7 16:53:32 2006 From: julien_lambreydesouza <@t> uqar.qc.ca (Julien Lambrey de Souza) Date: Tue Feb 7 16:54:38 2006 Subject: [Histonet] Celestine blue B Message-ID: <24a5b4a1ec0ded0269729de93511c01c@uqar.qc.ca> Hello. I have a protocol referecing Dury's and Wallington's book: Carleton's histological techniques. I have to do this protocol fast and don't have time to get access to the book. My protocol uses Mayer's haematoxylin and celestine blue B as used by these authors in the book. Would someone be kind enough to share the info. Thankyou, Julien Lambrey de Souza Research assistant, Evolutionary biology University of Quebec at Rimouski Tel: (418) 723-1986 #1714 Fax: (418) 724-1849 From craig.case <@t> thermo.com Tue Feb 7 17:26:55 2006 From: craig.case <@t> thermo.com (Case, Craig T.) Date: Tue Feb 7 17:27:05 2006 Subject: [Histonet] Attn Vendors Message-ID: Louri: We would be happy to supply you with prices and information if you can tell me where your client is located. Best Regards, Craig Craig T. Case Western Regional Manager Clinical Diagnostics Anatomical Pathology Thermo Electron Corporation 800-245-6212 ext. 4721 563-663-0428 (cell) 563-583-9244 (fax) craig.case@thermo.com From llewllew <@t> shaw.ca Tue Feb 7 18:09:09 2006 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Tue Feb 7 18:09:12 2006 Subject: [Histonet] Celestine blue B References: <24a5b4a1ec0ded0269729de93511c01c@uqar.qc.ca> Message-ID: <001a01c62c43$e23a9980$690a4246@yourlk4rlmsu> Go to http://stainsfile.info/StainsFile/stain/nuclei/celb_hem.htm it is there. Bryan Llewellyn ----- Original Message ----- From: "Julien Lambrey de Souza" To: Sent: Tuesday, February 07, 2006 2:53 PM Subject: [Histonet] Celestine blue B > Hello. > > I have a protocol referecing Dury's and Wallington's book: Carleton's > histological techniques. I have to do this protocol fast and don't have > time to get access to the book. > > My protocol uses Mayer's haematoxylin and celestine blue B as used by > these authors in the book. > > Would someone be kind enough to share the info. > > Thankyou, > > > Julien Lambrey de Souza > Research assistant, Evolutionary biology > University of Quebec at Rimouski > > Tel: (418) 723-1986 #1714 > Fax: (418) 724-1849 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jwatson <@t> gnf.org Tue Feb 7 18:19:14 2006 From: jwatson <@t> gnf.org (James Watson) Date: Tue Feb 7 18:19:32 2006 Subject: [Histonet] De Olmos Staining Message-ID: We have been doing the De Olmos sating for degenerating nerve fibers, but have run into a problem of the red blood cells picking up the silver stain. Has anyone had this problem and what did you do to correct it. Thank you. James Watson HT, ASCP Facilities Manager of Histology GNF, Genomics Institute of the Novartis Research Foundation Room C015 858-332-4647 jwatson@gnf.org From tahseen <@t> brain.net.pk Tue Feb 7 12:47:46 2006 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Tue Feb 7 20:50:35 2006 Subject: [Histonet] CISH Message-ID: <007701c62c16$fe656b80$972bfea9@m7c0y4> Hi All, I was wondering if anyone can let me know the simple steps involved in performing CISH . Thanks in advance, Muhammad Tahseen, MLT(Punjab Medical Faculty) MT(JIMTEF)Japan Histology Supervisor SKMCH & RC Pakistan. From L.Driessen <@t> orthop.umcn.nl Wed Feb 8 00:18:42 2006 From: L.Driessen <@t> orthop.umcn.nl (L.Driessen@orthop.umcn.nl) Date: Wed Feb 8 00:18:55 2006 Subject: [Histonet] RE: Technovit 7100 resin removal Message-ID: <2E2AC813A84E054C8E8E572131082BE214552C@umcnet14.umcn.nl> To my knowledge you can't!!! Perhaps there is a solvent that does remove the resin but I'm affraid that it will also distroy your tissue. I have never read or heard of anyone who can. Try MMA (like Technovit 9100) instead of GMA (Technovit 7100). l?on Driessen Orthopaedic Research Lab UMC St. Radboud, Nijmegen The Netherlands -----Oorspronkelijk bericht----- Van: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]Namens histonet-request@lists.utsouthwestern.edu Verzonden: dinsdag 7 februari 2006 19:22 Aan: histonet@lists.utsouthwestern.edu Onderwerp: Histonet Digest, Vol 27, Issue 10 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-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@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. RE: Immunofluorescence Text (Houston, Ronnie) 2. "MBS" in Luxol Fast Blue MBS (mlb@nmr.mgh.harvard.edu) 3. Poster on RE: Color coding (Gayle Callis) 4. NEW TISSUE PROCESSOR (Sue Barnes) 5. RE: "MBS" in Luxol Fast Blue MBS (Bartlett, Jeanine) 6. small dryer (Steven Coakley) 7. Re: small dryer (Maria Christensen) 8. Re: Immunohistochemistry on mouse teeth (Amy Porter) 9. RE: MSH6 (Barry Madigan) 10. RE: Immunohistochemistry on mouse teeth (Patsy Ruegg) 11. Re: "MBS" in Luxol Fast Blue MBS (John Kiernan) 12. CPT codes for muscle biopsies (Garrison, Becky) 13. Can anyone explain or at least empathise? (Cheryl) 14. Can anyone explain or at least empathise? (Cheryl) 15. Barcode Labeling on Tissue Slides (Gang Li) 16. Kangaroo Mammary Tissue (Bruce Abaloz) 17. RE: NEW TISSUE PROCESSOR (Anne Van Binsbergen) 18. RE: Can anyone explain or at least empathise? (Kemlo Rogerson) 19. Technovit 7100 resin removal (Jorge Tornero) 20. troubles with CDK4 (jhaviland@mdanderson.org) 21. Retic staining on rodent livers (Susan.Ferrigon@sanofi-aventis.com) 22. Job posting : Sales and Support opportunities at Improvision Inc (Pippa Simpson) 23. decalcifying before HBQ stain (Julien Lambrey de Souza) 24. JOB OPENING IN AUSTIN TEXAS (Chris Pomajzl) 25. Luxol dyes (Robert Krug) 26. re: decalcifying before HBQ stain (bhewlett@cogeco.ca) 27. RE: Decalcifying before HBQ (Julien Lambrey de Souza) 28. Technovit 7100 resin removal (germckeon@excite.com) ---------------------------------------------------------------------- Message: 1 Date: Mon, 6 Feb 2006 12:56:30 -0500 From: "Houston, Ronnie" Subject: RE: [Histonet] Immunofluorescence Text To: 'Patsy Ruegg' , 'David Haagsma' , histonet@lists.utsouthwestern.edu Message-ID: <00FBB1F3374BE24AAB7ACC6FCC7C617F561F4C@bsrexms01.bshsir.com> Content-Type: text/plain the best text I have come across dealing with Immunofluorescence is: Protein Localization by Fluorescence Microscopy: a practical approach ed. VJ Allan Oxford University Press 2000 ISBN 0-19-963741-5 (hbk) 0-19-963740-7 (pbk) Ronnie Houston Director of Anatomic Pathology Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 2877972 (804) 2877906 - fax ronnie_houston@bshsi.com -----Original Message----- From: Patsy Ruegg [mailto:pruegg@ihctech.net] Sent: Monday, February 06, 2006 12:51 PM To: 'David Haagsma'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Immunofluorescence Text The NSH IHC Resource Group has put together a list of reference books for IHC. I don't think there is specifically one on just F-IHC but several of them relate. You can join the IHCRG online at www.ihcrg.org for this and many other benefits relating to IHC. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of David Haagsma Sent: Monday, February 06, 2006 7:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Immunofluorescence Text Dear Histonet; We have a customer that is looking for a good immuno book. We currently do not offer a book of this type and I do not have any recommendations for her - can you help her out? Thank you, Dave Haagsma MT(ASCP) MarketLab Inc. Subject: Find Product Form name...........: Caroline Ghaleb suggestion.....: Dear Sir, You have a good number of educational book, unfortunately I'm looking to buy a book about immunofluorescence. I appreciate if you can let me know if you can provide such a book. Thanks a lot. Caroline Ghaleb phone..........: email..........: teknolab@rcn.com _______________________________________________ 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 ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone, and permanently delete the original e-mail, attachment(s), and any copies. ------------------------------ Message: 2 Date: Mon, 6 Feb 2006 13:47:25 -0500 (EST) From: mlb@nmr.mgh.harvard.edu Subject: [Histonet] "MBS" in Luxol Fast Blue MBS To: histonet@lists.utsouthwestern.edu Message-ID: <1146.132.183.203.6.1139251645.squirrel@mail.nmr.mgh.harvard.edu> Content-Type: text/plain;charset=iso-8859-1 Hi, I had tried sending this message upon joining histonet, but I didn't see it in the archive. That could be why I haven't gotten a reply yet! Here it is again: Hi, I've been using luxol fast blue MBS for myelin staining and would like to know what the acronym "MBS" represents. I have read many primary sources, such as articles by Kluver and Barrera as well as Salthouse, and I have performed internet searches with Google, all to no avail (other than finding histonet!). Thanks for your help! Megan =-) p.s. Also, any explanations of the other acronyms used in luxol dyes (ARN, G and the "N" in MBSN) would be much appreciated! ------------------------------ Message: 3 Date: Mon, 06 Feb 2006 11:59:27 -0700 From: Gayle Callis Subject: [Histonet] Poster on RE: Color coding To: "Horn, Hazel V" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20060206115815.01b4cab0@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Hazel, I remember your excellent poster, it was very informative. Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 ------------------------------ Message: 4 Date: Mon, 6 Feb 2006 14:34:53 -0500 From: "Sue Barnes" Subject: [Histonet] NEW TISSUE PROCESSOR To: "Histonet \(E-mail\)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" We are in the market for a new tissue processor. I am looking for one that is compatable with recycling of alcohol. Could I please hear from those of you who recycle alcohol and how well your processor works for you. Susan Barnes East Liverpool City Hospital East Liverpool, Ohio ------------------------------ Message: 5 Date: Mon, 6 Feb 2006 14:42:53 -0500 From: "Bartlett, Jeanine" Subject: RE: [Histonet] "MBS" in Luxol Fast Blue MBS To: , Message-ID: Content-Type: text/plain; charset="US-ASCII" Luxol fast blue MBSN is the chemical description for the specific dye, Solvent Blue 38 C32H12CuN8Na2O6S2 F.W. 732.16 Jeanine Bartlett, BS, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of mlb@nmr.mgh.harvard.edu Sent: Monday, February 06, 2006 1:47 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] "MBS" in Luxol Fast Blue MBS Hi, I had tried sending this message upon joining histonet, but I didn't see it in the archive. That could be why I haven't gotten a reply yet! Here it is again: Hi, I've been using luxol fast blue MBS for myelin staining and would like to know what the acronym "MBS" represents. I have read many primary sources, such as articles by Kluver and Barrera as well as Salthouse, and I have performed internet searches with Google, all to no avail (other than finding histonet!). Thanks for your help! Megan =-) p.s. Also, any explanations of the other acronyms used in luxol dyes (ARN, G and the "N" in MBSN) would be much appreciated! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Mon, 6 Feb 2006 12:04:07 -0800 (PST) From: Steven Coakley Subject: [Histonet] small dryer To: Histonet@lists.utsouthwestern.edu Message-ID: <20060206200407.41079.qmail@web90210.mail.scd.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Does anyone know where I might be able to get a small forced air dryer, holding 2-3 racks of slides? Steve --------------------------------- Brings words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. ------------------------------ Message: 7 Date: Mon, 6 Feb 2006 15:01:37 -0600 From: Maria Christensen Subject: Re: [Histonet] small dryer To: Steven Coakley , Message-ID: Content-Type: text/plain; charset="iso-8859-1" ; format="flowed" >Does anyone know where I might be able to get a >small forced air dryer, holding 2-3 racks of >slides? > > Steve > > >--------------------------------- >Brings words and photos together (easily) with > PhotoMail - it's free and works with Yahoo! Mail. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet We have one from TBS, Slide Dryer II. It works pretty well for what we use it (dry 130-160 slides O.N. at 37C) but it can dry slides in about half an hour. I think it lists for $1500 at Fisher Scientific, but we get a nice discount from them. Hope this helps! -- Mar?a Maria A. Christensen Technical Associate Dept. of Medical Microbiology & Immunology Creighton University School of Medicine 2500 California Plaza Omaha, Nebraska 68178-0404 voice (402) 280-2678 e-mail mariac@creighton.edu ------------------------------ Message: 8 Date: Mon, 6 Feb 2006 14:25:02 -0500 From: "Amy Porter" Subject: Re: [Histonet] Immunohistochemistry on mouse teeth To: "Horn, Diane A" , Message-ID: <001001c62b53$08344100$8e7a0923@HistoJJ> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Diane - Are you using a mouse primary? Didn't make a comment about this in your original post. If you are you would want to be using mouse on mouse staining techniques or kits. We always use another piece of tissue from the same animal if possible as a postive control, usually something from G.I. tract. Lots of proliferation going on there. Hope this helps out, ----- Original Message ----- From: "Horn, Diane A" To: Sent: Sunday, February 05, 2006 11:19 PM Subject: [Histonet] Immunohistochemistry on mouse teeth I am currently doing IHC on develping mouse teeth and I am getting only backgroung staining. The tissues are fixed O/N in 4% paraformaldehyde @4 degrees and embedded in paraffin. I have cut my sections @ 4um. I have tried HIER with Vector unmasking solution (boiling) for 2 minutes and using a kit for PCNA from Zymed. Has anyone done IHC on teeth that could give me some advice on what to try next? Thanks! Diane _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Tue, 7 Feb 2006 07:51:24 +1000 From: "Barry Madigan" Subject: RE: [Histonet] MSH6 To: Message-ID: <000001c62b67$7a6eb7a0$0201010a@WORKSTATION1> Content-Type: text/plain; charset="us-ascii" Hello Josie, The antibodies for mismatch repair genes that we use consists of MLH1, MSH2 and MSH6 all of which we obtain from BD (Becton Dickinson). They are all performed on our BOND MAX Immunostainer using the high pH retrieval solution with EDTA (30 min) and a Vision Polymer Detection System. We have no troubles with them except sometimes we find that with archival cases we need to apply extra heat for MLH1. When this extra step is performed the MLH1 internal controls come up beautifully. Before we had the BOND MAX we used to heat retrieve using a microwave pressure cooker for 8 minutes on high in TRIS/EDTA pH 9.0 for MLH1. For the MSH6 antibody we currently obtain a dilution of 1:200. MSH2 1:500 and MLH1 1:100. Hope this is of some help. Regards Barry Barry Madigan Immunohistochemistry QHPS-Royal Brisbane Hospital Campus Australia -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Orr, Rebecca Sent: Saturday, 4 February 2006 1:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] MSH6 Hi Josie, How many different types of protocols did you try on the XT? With the different temp and Cell conditioning options I'm surprised you didn't get something. Are you running the MLH-1 and MSH-2 as well? How are they? My experience with the Microsateliite Instabilities is 4+ staining in 2 out of 3 antibodies, depending on the control tissue. The third antibody would compare at 2+ or a bit stronger. I have the best of both worlds with a Benchmark and a Nemesis. I experienced the poor staining results of these particular antibodies and I decided to run them on the Nemesis with Biocare polymer detection and they are gorgeous. I use the Biocare antibodies, too. In case you don't have that option, I would consider trying another control tissue, and running through all the CC options. I'm not sure if they behave differently without heat, did you try running a protocol with out heat in the detection? Do you have access to the polymer from Ventana? Maybe try that? Or run the antibody at 1:5 or 1:10. Or do you have a pressure cooker to try the HEIR offline. If you get good staining with offline HEIR then you can rule out the detection as the issue, too. Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 -----Original Message----- Message: 8 Date: Wed, 1 Feb 2006 13:54:36 -0600 From: "Nava, Josefa" Subject: [Histonet] looking for Antibodies-MSH6 and PMS2 To: Message-ID: <2C515C1049EAF5459EFD8C9B929078A41945A8@phdex03.txhealth.org> Content-Type: text/plain; charset="us-ascii" Hello Everyone , I am using Ventana BMK/XT , can someone tell me a good source of MSH6 and PMS2 that will work on the Ventana Machine. I appreciate any information. Thank you. Josie _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 10 Date: Mon, 6 Feb 2006 15:05:30 -0700 From: "Patsy Ruegg" Subject: RE: [Histonet] Immunohistochemistry on mouse teeth To: "'Amy Porter'" , "'Horn, Diane A'" , Message-ID: <200602062205.k16M5NDZ009781@chip.viawest.net> Content-Type: text/plain; charset="US-ASCII" Have you decalcified the mouse teeth (I would imagine that you did)? How was the performed? Decal can have a determental effect on IHC. I use formic acid for good IHC results. If the calcium is not removed it can block access to the antigen site. Just some thoughts. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Porter Sent: Monday, February 06, 2006 12:25 PM To: Horn, Diane A; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Immunohistochemistry on mouse teeth Diane - Are you using a mouse primary? Didn't make a comment about this in your original post. If you are you would want to be using mouse on mouse staining techniques or kits. We always use another piece of tissue from the same animal if possible as a postive control, usually something from G.I. tract. Lots of proliferation going on there. Hope this helps out, ----- Original Message ----- From: "Horn, Diane A" To: Sent: Sunday, February 05, 2006 11:19 PM Subject: [Histonet] Immunohistochemistry on mouse teeth I am currently doing IHC on develping mouse teeth and I am getting only backgroung staining. The tissues are fixed O/N in 4% paraformaldehyde @4 degrees and embedded in paraffin. I have cut my sections @ 4um. I have tried HIER with Vector unmasking solution (boiling) for 2 minutes and using a kit for PCNA from Zymed. Has anyone done IHC on teeth that could give me some advice on what to try next? Thanks! Diane _______________________________________________ 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 ------------------------------ Message: 11 Date: Mon, 06 Feb 2006 17:12:53 -0500 From: John Kiernan Subject: Re: [Histonet] "MBS" in Luxol Fast Blue MBS To: mlb@nmr.mgh.harvard.edu Cc: histonet@lists.utsouthwestern.edu Message-ID: <43E7C9E5.B2FC7F08@uwo.ca> Content-Type: text/plain; charset=us-ascii Your message appeared all right, but nobody answered. It probably means nobody knows what the MBS stands for. Luxol fast blues are not a chemical group of dyes. They are arylguanidinium salts of anionic dyes (rather than the usual sodium salts). This makes the dyes soluble in alcohol but not in water. When sections are stained, the organic cation stays in the alcohol, and the coloured anion lodges in the rather myelin. Like any other anionic dye, a luxol can be differentiated by alkali. Luxol fast blue MBS is a phthalocyanine dye; luxol fast blues G and ARN are azo dyes. See Clasen et al. 1973 J. Neuropath. Exp. Neurol. 32:271-283 for more on this subject, including how to make your own luxol-type dyes. John Kiernan London, Canada ----------------------------- mlb@nmr.mgh.harvard.edu wrote: > > Hi, I had tried sending this message upon joining > histonet, but I didn't see it in the archive. That > could be why I haven't gotten a reply yet! Here it > is again: > > Hi, I've been using luxol fast blue MBS for myelin > staining and would like to know what the acronym "MBS" > represents. > > I have read many primary sources, such as articles by > Kluver and Barrera as well as Salthouse, and I have > performed internet searches with Google, all to no > avail (other than finding histonet!). > > Thanks for your help! > Megan > =-) > > p.s. Also, any explanations of the other acronyms used > in luxol dyes (ARN, G and the "N" in MBSN) would > be much appreciated! > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Mon, 6 Feb 2006 17:14:00 -0500 From: "Garrison, Becky" Subject: [Histonet] CPT codes for muscle biopsies To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Could any of you share the CPT codes used for muscle biopsy work ups? My question is in 2 parts: 1) We currently charge: 88314 per stain for ATP, NADH, Esterase, Alkaline Phosphatase, Acid Phosphatase, AMPDA, Myophorylase, Cyto C Oxidase, and SDH on snap frozen muscle tissue and 88313 per stain for Trichrome, PAS, Oil Red O (ORO) on snap frozen muscle tissue in addition to 88305 for the biopsy itself. One pathologist is suggesting that we should be charging 88319 for the ATP thru SDH above and 88314 for the Trichrome thru ORO above and that 88313 to be used only on Trichromes, PAS and ORO on paraffin embedded tissue (such as for the nerve biopsies). In reviewing the 2006 CPT code book, it does seem that we may be charging incorrectly. 2) If 88319 is the correct answer to the first part: 88319 is not listed as an add on code. Do you still charge an 88305 for the biopsy? How would you charge a muscle biospsy, received fresh, divided into 2 parts as follows: First part is snap frozen in isopentane; frozen sections are stained with ATP x 3, SDH, Trichrome, ORO. Second part is submitted for routine processing and paraffin sections are stained with H&E, Trichrome and ORO? Thanks for your input. Becky Garrison Pathology Supervisor Jacksonville, FL 904-244-6237 ------------------------------ Message: 13 Date: Mon, 6 Feb 2006 16:01:57 -0800 (PST) From: Cheryl Subject: [Histonet] Can anyone explain or at least empathise? To: histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Message-ID: <20060207000157.95181.qmail@web50913.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hey Fellow Histonetters: I read all the postings on here and generally understand all of it, but the more I read the more I realize I've forgotten more than I probably remember. (I haven't done a muscle stain group in quite a while!!). Does anyone else feel this way??? (Yes, I've passed 40, I'm sure someone out there can explain the mechanism of memory loss for me!!!) Thanks for slowing down the brain drain!! Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one tech at a time. admin@fullstaff.org 281.852.9457 office 281.883.7704 cell ------------------------------ Message: 14 Date: Mon, 6 Feb 2006 16:01:57 -0800 (PST) From: Cheryl Subject: [Histonet] Can anyone explain or at least empathise? To: histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Message-ID: <20060207000157.95181.qmail@web50913.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hey Fellow Histonetters: I read all the postings on here and generally understand all of it, but the more I read the more I realize I've forgotten more than I probably remember. (I haven't done a muscle stain group in quite a while!!). Does anyone else feel this way??? (Yes, I've passed 40, I'm sure someone out there can explain the mechanism of memory loss for me!!!) Thanks for slowing down the brain drain!! Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one tech at a time. admin@fullstaff.org 281.852.9457 office 281.883.7704 cell ------------------------------ Message: 15 Date: Mon, 6 Feb 2006 19:36:09 -0500 From: Gang Li Subject: [Histonet] Barcode Labeling on Tissue Slides To: histonet@lists.utsouthwestern.edu Message-ID: <120b7bf0602061636l18f78fb7m315a28e5c321e5c6@mail.gmail.com> Content-Type: text/plain; charset=ISO-8859-1 Could anybody kindly tell me the barcode standards used for tissue slides labeling? Best wishes Gang Li, Ph.D. Biomedical Photometrics Inc. Waterloo, Ontario, CANADA ------------------------------ Message: 16 Date: Tue, 07 Feb 2006 13:14:52 +1100 From: Bruce Abaloz Subject: [Histonet] Kangaroo Mammary Tissue To: histoNet@Pathology.swmed.edu Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" Dear Histonetter's, I am looking for stains that will identify the basic contents of cultured, mammary alveoli (mammospheres) of Kangaroo. Broad lipid ( ANY FAT STAIN THAT CAN BE DONE ON PARAFFIN EMBEDDED TISSUE???POSSIBLE??), carbohydrate (PAS?? ANY OTHER SUGGESTIONS) and protein stains (???) are required to determine what - IF any, different hormone combinations have/ effect on milk production. Any suggestions would be wonderful......our Histologist has made several good suggestions for me to try & we are both interested to see 'WHAT' POSSIBILITIES/suggestions the scientific minds in Histo-land come up with. Thankyou in advance & Cheers, Sonia -- BRUCE ABALOZ PH:61383446282 HISTOLOGIST FAX:61383447909 DEPT.of ZOOLOGY EMAIL: brucea@unimelb.edu.au THE UNIVERSITY Of MELBOURNE. VICTORIA. AUSTRALIA 3010 Nobody Can Make You Feel Inferior Without YOUR Permission - Eleanor Roosevelt DANCE LIKE NO-ONE'S WATCHING Q: What's a specimen? A: An Italian astronaut. ******************************************************************************** This email and any attachments may contain personal information or information that is otherwise privileged, confidential or otherwise protected from disclosure/subject of copyright. Any use, disclosure or copying of any part of it is prohibited. The University does not warrant that this email or any attachments are free from viruses or defects. Please check any attachments for viruses and defects before opening them. If this email is received in error please delete it and notify us by return email. The University does not necessarily share or endorse the views expressed in this email. ------------------------------ Message: 17 Date: Tue, 7 Feb 2006 08:11:12 +0400 From: "Anne Van Binsbergen" Subject: RE: [Histonet] NEW TISSUE PROCESSOR To: "Sue Barnes" , "Histonet (E-mail)" Message-ID: <7E070A7B959A9F42BE732545E5CF6210948D4D@SKMCEMAIL.skmc.gov.ae> Content-Type: text/plain; charset="us-ascii" We use recycled alcohol and have a Sakura VIP5 - in my opinion they are the best - no problems whatsoever. Recycled alcohol should not make any difference. Annieinarabia -----Original Message----- From: Sue Barnes [mailto:SBarnes@elch.org] Sent: Monday, February 06, 2006 11:35 PM To: Histonet (E-mail) Subject: [Histonet] NEW TISSUE PROCESSOR We are in the market for a new tissue processor. I am looking for one that is compatable with recycling of alcohol. Could I please hear from those of you who recycle alcohol and how well your processor works for you. Susan Barnes East Liverpool City Hospital East Liverpool, Ohio _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Tue, 7 Feb 2006 08:48:32 -0000 From: Kemlo Rogerson Subject: RE: [Histonet] Can anyone explain or at least empathise? To: "'Cheryl'" , histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain I've rationalised from the following standpoint; When you are younger you usually have less to think about and therefore you can focus your thoughts better. As you get older you have more to think about and usually less time and your thoughts become unfocused. I mean my son arranged perfectly easily to organise his snow boarding holiday but fails regularly to pay his rent. I think when you get older you also care more about forgetting; damn where are my glasses!!! Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Cheryl [mailto:tkngflght@yahoo.com] Sent: Tuesday, February 07, 2006 12:02 AM To: histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: [Histonet] Can anyone explain or at least empathise? Hey Fellow Histonetters: I read all the postings on here and generally understand all of it, but the more I read the more I realize I've forgotten more than I probably remember. (I haven't done a muscle stain group in quite a while!!). Does anyone else feel this way??? (Yes, I've passed 40, I'm sure someone out there can explain the mechanism of memory loss for me!!!) Thanks for slowing down the brain drain!! Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one tech at a time. admin@fullstaff.org 281.852.9457 office 281.883.7704 cell _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 19 Date: Tue, 7 Feb 2006 13:32:52 +0100 From: Jorge Tornero Subject: [Histonet] Technovit 7100 resin removal To: histonet@lists.utsouthwestern.edu Message-ID: <8c964a790602070432r457655a2x@mail.gmail.com> Content-Type: text/plain; charset=ISO-8859-1 Hi all, I would like to remove the resin in my slides prior to stain. Do you know any protocol to do this? Thank you very much in advance Jorge Tornero IEO C?diz - Spain ------------------------------ Message: 20 Date: Tue, 7 Feb 2006 08:43:35 -0600 From: jhaviland@mdanderson.org Subject: [Histonet] troubles with CDK4 To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Hello: I am trying to do IHC with rabbit anti human CDK4 (D-22) from Santa Cruz on salivary gland tissue. I am having background problems which is leading to difficulty in figuring out what is positive and what is negative. I have used LSAB2 kit from Dako, Rabbit Envision Polymer from Dako and the Rabbit ABC Elite kit from Vector. I do a 3% hydrogen peroxide treatment for 20 minutes and do antigen retrieval with 10mM Citric Buffer pH 6.0 for 45 minutes in a steamer and a 20 minute cool down. I do a blocking step and have used either serum free or goat serum for a block. I already checked with the company about my protocol and it is the same as theirs. I tried doing it on skin which is reccommended as a control but still have background problems. Thanks Joie Haviland Senior Research Assistant MD Anderson Cancer Research Center ------------------------------ Message: 21 Date: Tue, 7 Feb 2006 14:53:48 +0000 From: Susan.Ferrigon@sanofi-aventis.com Subject: [Histonet] Retic staining on rodent livers To: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Does anyone have problems with staining for reticulin on rodent livers?? if not, do you have a particular method you use?? I use Gordon and Sweet and find the fibres don't stain up very well and the background is grainy. Any advise. Thanks Susan ------------------------------ Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de ce courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuellement pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ------------------------------ ------------------------------ Message: 22 Date: Tue, 7 Feb 2006 15:04:07 -0000 From: "Pippa Simpson" Subject: [Histonet] Job posting : Sales and Support opportunities at Improvision Inc To: Message-ID: <017d01c62bf7$bedc6870$0701a8c0@improvision.com> Content-Type: text/plain; charset="iso-8859-1" Please forgive the commercial intrusion. Improvision is currently considering applicants for roles within our Sales and Technical Support teams. For further details please visit http://www.improvision.com/careers/ or contact recruitment@improvision.com. Please do not reply to the listserver. Kind regards Pippa Simpson Improvision Human Resources ------------------------------ Message: 23 Date: Tue, 7 Feb 2006 10:50:25 -0500 From: Julien Lambrey de Souza Subject: [Histonet] decalcifying before HBQ stain To: histonet@lists.utsouthwestern.edu Message-ID: <75c6cee121c47a552bf01aa9b6910f1b@uqar.qc.ca> Content-Type: text/plain; charset=US-ASCII; format=flowed Hello everyone, Just a quick question: Is it necessary to decalcify before staining with the HBQ protocol? Thanks, Julien Lambrey de Souza Research assistant, Evolutionary biology University of Quebec at Rimouski Tel: (418) 723-1986 #1714 Fax: (418) 724-1849 ------------------------------ Message: 24 Date: Tue, 7 Feb 2006 10:12:11 -0600 From: "Chris Pomajzl" Subject: [Histonet] JOB OPENING IN AUSTIN TEXAS To: Message-ID: <006901c62c01$43393590$26fca8c0@CSP> Content-Type: text/plain; charset="iso-8859-1" Greetings: We have multiple shift openings in Austin, Texas. If you might be interested in a fast-paced, high-volume laboratory, please give me a call. Duties include embedding, sectioning, special stains, etc. Competetive salary, health benefits, and a friendly working environment. Thanks. Sincerely, Chris Pomajzl, HTL (ASCP) Histology Supervisor Clinical Pathology Laboratories, Inc. 512.873.1660 (o) 512.873.5004 (f) cpomajzl@cpllabs.com CONFIDENTIALITY NOTICE: This communication is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged, confidential, and exempt from disclosure under applicable law. If you are not the intended recipient, you are notified that any use, dissemination, distribution, or copying of this communication is strictly prohibited. If you have received this message in error, please notify the sender immediately and destroy this communication. ------------------------------ Message: 25 Date: Tue, 7 Feb 2006 10:17:43 -0600 From: Robert Krug Subject: [Histonet] Luxol dyes To: histonet@lists.utsouthwestern.edu Cc: mlb@nmr.harvard.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Megan: Saw Dr Kiernan's posting on Histonet. I cannot tell you what the MBS stands for, but I can add a little insight into naming dyes. Dyes are given trivial names in most cases. This really means the person originating the dye may call the dye whatever they please. At one time there were may have existed guidelines. If such guidelines existed, no one seems to be familiar with the specifics at this late date. Today we use scientific nomenclature with set conventions to name products such as 1-Cyclohexyl-2-phyrrolidone. Personally I prefer the trivial names. In some cases the abbreviations come from the German dye industry - who were really pioneers in the field. The SF in Light Green SF yellowish = SaureFarbstoff (German) = acid dye English terms are also sometimes abbreviated FCF in Fast Green FCF = For Coloring Food There are a series of fluorescent dyes available such as PKH26, PKH67. In this particular case I cannot be 100% sure, but the person who filed the patent on these dyes just happened to have the initials PKH. Coincidence? In this particular case I would guess that the 26th and the 67th attempts were keepers. Naphthol compounds are often named Naphthol AS-MX phosphate or possibly Naphthol AS-BI phosphate. The structures are evidently different, but no one I have talked to has been able to explain why one product is given as the AS-MX designation and the next product is AS-BI If you look at Conn's Biological Stains, 9th Edition, you will find the text lists many more obscure synonyms - as compared to the 10th Edition. In previous years, companies often gave common dyes unique names. They may have believed they gained some competitive advantage from this practice. Even today dyes may often go by more than one name. Sometime this is done when the company markets to various industries. If you are selling to the printing industry, you might name a dye XXXX. If you market dyes for histological use, you might name a dye YYYY. Today I believe the trend is for companies to sell the dye by the name given the most common product of commerce. This is probably why fewer synonyms are listed in the 10th Edition. Synonyms however are here to stay. Sodium thiosulfate is still referred to as hypo in older textbooks. This is because "hypo" was the term used in photography. One of the many useful functions served by the Biological Stain Commission was the creation of CI or Color Index numbers. It really doesn't matter what a company names a dye. If Product X and Product Y from competing companies have the same CI number, there is a good chance the dyes may be used in the same staining procedures. If the dyes are Certified by the BSC, this gives even more indication the dye should be acceptable for use. The CI number varies from the CAS number. For the CAS numbers to be identical, the molecular structure should be identical. Not so with Color Index numbers. For example, Basic Fuchsin may be composed of 100% Pararosaniline, or a mixture of pararosaniline, rosaniline, magenta II and magenta III. The pararosaniline may be the chloride or the acetate form. So although the blends may vary from manufacturer to manufacturer and possibly even between lot numbers for a specific manufacturer, the dye must perform in an acceptable manner to receive certification from the Biological Stain Commission. In the 10th Edition of Conn's, no CI number is assigned to Basic Fuchsin, although the individual homologs are given given unique homologs. The 9th Edition of Conn's assigned Basic Fuchsin the CI number for Pararosanline. The real test is how well the dyes perform in actual staining procedures. Anyone really interested in dyes for biological use should try to obtain a copy of the methods used by the Biological Stain Commission. Analysis and testing of biological stains - The Biological Stain Commission Procedures (Biotechnic & Histochemistry 2002, 77(5&6): 237-275 Methylene Blue is another prime example for a dye which is composed of a series of closely related homologs. However in this case Methylene Blue is assigned a Color Index number, as opposed to Basic Fuchsin which was not assigned a unique CI number in the 10th Edition of Conn's. Again the major homologs are assigned CI numbers. Other dyes by comparison are relatively pure and may be almost 100% pure. However sometimes purity is not all. Some of the impurities may be present/added/act as stabilizers. In the case of Nile Blue, the presence of Nile Red as a contaminant may be highly desirable. For other dyes the formulations are constantly shifting, even though the name remains relatively constant. Alcian Blue is a dye which has a tendency to explode during manufacture. This has resulted in a constant tinkering with the method of production over the years. The dye sold today was probably manufactured very differently than an Alcian Blue manufactured decades ago. If the product of commerce continues to provide acceptable staining results in the methods commonly associated with Alcian Blue, the dye will be sold & certified as Alcian Blue. It would be impractical to give each slight variation in product a unique name or CAS number. Trying to perform a literature search would be made extremely difficult to almost impossible. Tradition methyl green has not been produced commercially for many years (35-45+ ?). The actual product of commerce today is ethyl green. In this case methyl green and ethyl green are really not synonyms. However because ethyl green may be used for the same purposes as methyl green, the product of commerce continues to be called methyl green - very much to the annoyance of Dr Kiernan :) Hope this helps, Best Regards, Bob Krug Sigma-Aldrich St Louis, MO ------------------------------ Message: 26 Date: Tue, 07 Feb 2006 11:37:48 -0500 From: bhewlett@cogeco.ca Subject: re: [Histonet] decalcifying before HBQ stain To: Julien Lambrey de Souza , histonet@lists.utsouthwestern.edu Message-ID: <43e8ccdc.319.3f3d.9906@cogeco.ca> > Julien, Just what is the HBQ protocol? Bryan > Hello everyone, > > Just a quick question: Is it necessary to decalcify before staining > with the HBQ protocol? > > Thanks, > > > Julien Lambrey de Souza > Research assistant, Evolutionary biology > University of Quebec at Rimouski > > Tel: (418) 723-1986 #1714 > Fax: (418) 724-1849 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 27 Date: Tue, 7 Feb 2006 12:50:31 -0500 From: Julien Lambrey de Souza Subject: [Histonet] RE: Decalcifying before HBQ To: histonet@lists.utsouthwestern.edu Message-ID: <634240d185312d7b6108d64157d0b1e0@uqar.qc.ca> Content-Type: text/plain; charset=US-ASCII; format=flowed Sorry. I guess I should have elaborated a bit on the stain. HBQ = Hall and Brunt's Quadruple stain Used to differenciate bone from cartillage and chondroid bone. Deparaffinized sections are stained in aqueous celestine blue (0,5% in 5% ammonium sulphate and 14% glycerin), Mayer's hematox, alcian blue (1% in 1% acetic acid, pH 2,6), Phosphomolybdic acid1% and aqueous direct red 0,5%. All the protocols and pictures I have seen in publications are done on decalcified tissue. My question is the following: If my specimen cuts out nice sections without decalcification, will I get the intended staining (same as seen in publications) or will it be different from decalcified sections? How does decalcification influence staining results? Thanks for the imput. Julien Lambrey de Souza Research assistant, Evolutionary biology University of Quebec at Rimouski Tel: (418) 723-1986 #1714 Fax: (418) 724-1849 ------------------------------ Message: 28 Date: Tue, 7 Feb 2006 12:52:02 -0500 (EST) From: "germckeon@excite.com" Subject: [Histonet] Technovit 7100 resin removal To: jorge.tornero@gmail.com Cc: histonet@lists.utsouthwestern.edu Message-ID: <20060207175202.C3E6137628@xprdmailfe14.nwk.excite.com> Content-Type: text/plain; charset="us-ascii" Hello Jorge, I am also trying to find out the same thing. Would it be possible to send me any hints you may get? Many thanks, Gerald McKeon Trinity College, Dublin, Ireland. [Histonet] Technovit 7100 resin removal Hi all, I would like to remove the resin in my slides prior to stain. Do you know any protocol to do this? Thank you very much in advance Jorge Tornero IEO C?diz - Spain _______________________________________________ Join Excite! - http://www.excite.com The most personalized portal on the Web! ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 27, Issue 10 **************************************** From carl.hobbs <@t> kcl.ac.uk Wed Feb 8 08:07:58 2006 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Wed Feb 8 08:08:43 2006 Subject: [Histonet] re: Technovit 7100 resin removal Message-ID: <001201c62cb9$10d89880$112b5c9f@Carlos> In all my years, I have never been able to remove the resin ( of ANY GMA-based resin) and yet still retain the section fixed to a slide, sigh! I bought the same Technovit resin for immunohistochemical applications yet have never found it to be a viable alternative to pwax sections. Methyl methac is the only resin that can be removed, IMHO, still retaining the section on the slide. Carl From SAllen <@t> exchange.hsc.mb.ca Wed Feb 8 08:34:14 2006 From: SAllen <@t> exchange.hsc.mb.ca (Sharon Allen) Date: Wed Feb 8 08:35:34 2006 Subject: [Histonet] Celestine blue B Message-ID: <36FE435D6D2F1D489174B22362A961B66B8313@hscxntmx0006.hsc.mb.ca> Hi, I started using Celestine Blue in our method for Gomori's Trichrome & it made a great improvement. I'm not sure why! Put the slides in Celestine Blue for 2 min. before the Mayer's Haematoxylin. Recipe is: Ferric ammonium sulfate...........5.0 gm Dissolve in distilled water.......100 ml. Celestine blue....................0.5 gm Boil for 3 min. to dissolve, filter, when cooled add- Glycerin..........................14 ml Stable for at least 6 months (Carleton's Histological Techniques 4th edition P.133) Hope this helps. Sharon sallen@hsc.mb.ca -----Original Message----- This e-mail and/or any documents in this transmission is intended for the address(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. From TillRenee <@t> uams.edu Wed Feb 8 08:43:26 2006 From: TillRenee <@t> uams.edu (Till, Renee) Date: Wed Feb 8 08:43:52 2006 Subject: [Histonet] argyrophil and argentaffin Message-ID: <11F927674DEBDC43B960809A7403C5D232866C@MAILPED.ad.uams.edu> I had heard that if you are doing a Grimelius for argyrophil cells, that an argentaffin stain should also be performed to be able to differentiate between the two cells. Is this true for any argyrophil stain? Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 ================================================================================================ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ================================================================================================ From bhewlett <@t> cogeco.ca Wed Feb 8 09:13:41 2006 From: bhewlett <@t> cogeco.ca (bhewlett@cogeco.ca) Date: Wed Feb 8 09:14:37 2006 Subject: [Histonet] argyrophil and argentaffin Message-ID: <43ea0aa5.322.b62.22262@cogeco.ca> > Renee, All argentaffin cells are also argyrophil by definition. Therefore, to distinguish the true argyrophil population, one should perform both stains and subtract the argentaffin population from the argyrophil. Bryan > I had heard that if you are doing a Grimelius for argyrophil cells, that > an argentaffin stain should also be performed to be able to > differentiate between the two cells. Is this true for any argyrophil > stain? > > > > Renee' Till, HT > > > > Arkansas Childrens Nutrition Center > > 1120 Marshall > > Little Rock, AR > > 72202 > > > > > > > =============================================================================================== > Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended > recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or > distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and > destroy all copies of the original message. > ===============================================================================================_______________________ > ________________________ Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bhewlett <@t> cogeco.ca Wed Feb 8 09:15:24 2006 From: bhewlett <@t> cogeco.ca (bhewlett@cogeco.ca) Date: Wed Feb 8 09:15:33 2006 Subject: [Histonet] argyrophil and argentaffin Message-ID: <43ea0b0c.40.65c1.3974@cogeco.ca> > Renee, All argentaffin cells are also argyrophil by definition. Therefore, to distinguish the true argyrophil population, one should perform both stains and subtract the argentaffin population from the argyrophil. Bryan > I had heard that if you are doing a Grimelius for argyrophil cells, that > an argentaffin stain should also be performed to be able to > differentiate between the two cells. Is this true for any argyrophil > stain? > > > > Renee' Till, HT > > > > Arkansas Childrens Nutrition Center > > 1120 Marshall > > Little Rock, AR > > 72202 > > > > > > > =============================================================================================== > Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended > recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or > distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and > destroy all copies of the original message. > ===============================================================================================_______________________ > ________________________ Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Samuel.Jones2 <@t> med.va.gov Wed Feb 8 09:08:12 2006 From: Samuel.Jones2 <@t> med.va.gov (Samuel.Jones2@med.va.gov) Date: Wed Feb 8 09:25:56 2006 Subject: [Histonet] Alcohol Disposal Message-ID: <476BAB689BEA3E4DBE6F8D5A90CAC2CF056FC7DA@vhantxexc1.v17.med.va.gov> Hello All: I've been asked by my peers to see how labs are collecting and disposing of their waste alcohol. Thoughts please. Thanks Samuel E. Jones, MS, HT(ASCP)HTL, QIHC Supervisor, Anatomic Pathology VA North Texas Health Care System 4500 South Lancaster Road, 113 Dallas, Texas 75216 Phone: 214-857-0659 e-mail: samuel.jones2@med.va.gov From Barry.R.Rittman <@t> uth.tmc.edu Wed Feb 8 09:42:40 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Wed Feb 8 09:42:43 2006 Subject: [Histonet] Celestine blue B Message-ID: We used to use this formula for general staining with Celestine blue but came across a formula in the literature many years ago that gave us a much more intense stain and also lasted a much longer time. Sorry but cannot remember the reference. >From memory so hope that this is correct. 0.5 gms Celestine blue powder into a glass beaker. Under the fume hood add 0.5 ml of concentrated sulfuric acid and mix with a glass rod into a paste. During this time the mixture bubbles and gives off fumes hence the fume hood. Add 100 ml of 5% aqueous iron alum containing 14 ml of glycerin. Mix well and filter. This solution stains very rapidly to start with and staining time decreases a little after that so you need to test it on sections for half a minute when it is first made. A good way to test whether Celestine blue stains are past their useful staining life is to swirl the solution around the bottle or flask and if the solution fails to stick somewhat to the glass then need to make up a fresh batch. Filter each time used. We really liked this modification as the nuclear stain when using van Gieson stain as it is not differentiated by the picric acid. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sharon Allen Sent: Wednesday, February 08, 2006 8:34 AM To: 'Julien Lambrey de Souza'; Histonet (E-mail) Subject: RE: [Histonet] Celestine blue B Hi, I started using Celestine Blue in our method for Gomori's Trichrome & it made a great improvement. I'm not sure why! Put the slides in Celestine Blue for 2 min. before the Mayer's Haematoxylin. Recipe is: Ferric ammonium sulfate...........5.0 gm Dissolve in distilled water.......100 ml. Celestine blue....................0.5 gm Boil for 3 min. to dissolve, filter, when cooled add- Glycerin..........................14 ml Stable for at least 6 months (Carleton's Histological Techniques 4th edition P.133) Hope this helps. Sharon sallen@hsc.mb.ca -----Original Message----- This e-mail and/or any documents in this transmission is intended for the address(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. From gentras <@t> vetmed.auburn.edu Wed Feb 8 09:47:36 2006 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Wed Feb 8 09:47:50 2006 Subject: Fwd: Re: [Histonet] Alcohol Disposal Message-ID: <6.0.1.1.0.20060208094734.01a81e48@mailhost.vetmed.auburn.edu> >Date: Wed, 08 Feb 2006 09:47:25 -0600 >To: Samuel.Jones2@med.va.gov >From: "Atoska S. Gentry" >Subject: Re: [Histonet] Alcohol Disposal > > >Hello, our campus' Risk Management Department picks up our waste Alcohol >along with other chemical waste. I'm not sure of their means of disposal. >Best wishes. Atoska > > >At 09:08 AM 2/8/2006, you wrote: >> >> >>Hello All: >> >> >> >>I've been asked by my peers to see how labs are collecting and disposing of >>their waste alcohol. Thoughts please. Thanks >> >> >> >>Samuel E. Jones, MS, HT(ASCP)HTL, QIHC >> >>Supervisor, Anatomic Pathology >> >>VA North Texas Health Care System >> >>4500 South Lancaster Road, 113 >> >>Dallas, Texas 75216 >> >>Phone: 214-857-0659 >> >>e-mail: samuel.jones2@med.va.gov >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sjchtascp <@t> yahoo.com Wed Feb 8 10:18:09 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Wed Feb 8 10:18:23 2006 Subject: [Histonet] backup cryostat Message-ID: <20060208161809.48345.qmail@web90208.mail.scd.yahoo.com> Our company is looking for a backup cryostat. Steve --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! From Malcolm.McCallum <@t> tamut.edu Wed Feb 8 10:39:25 2006 From: Malcolm.McCallum <@t> tamut.edu (Malcolm McCallum) Date: Wed Feb 8 10:40:27 2006 Subject: [Histonet] infrared vs uv in sequencing Message-ID: anyone familiar with the advantages/disadvantages? Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Till, Renee Sent: Wed 2/8/2006 8:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] argyrophil and argentaffin I had heard that if you are doing a Grimelius for argyrophil cells, that an argentaffin stain should also be performed to be able to differentiate between the two cells. Is this true for any argyrophil stain? Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 ================================================================================================ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ================================================================================================ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tkngflght <@t> yahoo.com Wed Feb 8 10:52:15 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Wed Feb 8 10:52:27 2006 Subject: [Histonet] Alcohol Disposal--a few choices In-Reply-To: <6.0.1.1.0.20060208094734.01a81e48@mailhost.vetmed.auburn.edu> Message-ID: <20060208165216.21889.qmail@web50902.mail.yahoo.com> Hi Samuel- This kind of issue usually starts with the water department in your district. Some places have enough water going through their system that this kind of waste can go down the drain. It is considered safe and cost effective ($0!!) because of the dilution factors. Hospitals use HUGE quantities of water so they might even make an exception if it's not in the base policies. Before you contact your water district, talk with your risk management and environmental services people--they may already know or might want to talk to the water people for you. If you end up not being able to dump it, (and the water company can usually answer you within a few days) you may want to consider recycling. BR and one other company (??) can do a cost analysis with just a few questions to determine if this won't SAVE your department a buncha bucks (including your people's time in the equasion.) These systems are SO effective the end result is sometimes better than the original solutions you got from your suppliers. You can do just alcohols or both Alc. and Xylenes. These systems are infinitely more useable and safer than they were 20 years ago. The most expensive options are usually waste haulers and they will charge you more for certain contaminants. Xylene, biologicals, etc. So when you get a quote from a waste hauler or incinerator, ask if they are going to recycle it themselves (should get you a lower price) or burn it, and any additional cost for other cross-contaminants from your stain line or processors. Being a tree-hugger from way back, I always chose the recycle method when it is fiscally responsible (and it usually is). Hope this helps! Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one tech at a time. admin@fullstaff.org 281.852.9457 o 281.883.7704 c "Atoska S. Gentry" wrote: >Date: Wed, 08 Feb 2006 09:47:25 -0600 >To: Samuel.Jones2@med.va.gov >From: "Atoska S. Gentry" >Subject: Re: [Histonet] Alcohol Disposal > > >Hello, our campus' Risk Management Department picks up our waste Alcohol >along with other chemical waste. I'm not sure of their means of disposal. >Best wishes. Atoska > > >At 09:08 AM 2/8/2006, you wrote: >> >> >>Hello All: >> >> >> >>I've been asked by my peers to see how labs are collecting and disposing of >>their waste alcohol. Thoughts please. Thanks >> >> >> >>Samuel E. Jones, MS, HT(ASCP)HTL, QIHC >> >>Supervisor, Anatomic Pathology >> >>VA North Texas Health Care System >> >>4500 South Lancaster Road, 113 >> >>Dallas, Texas 75216 >> >>Phone: 214-857-0659 >> >>e-mail: samuel.jones2@med.va.gov >> >> >> >>_______________________________________________ >>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 From meint002 <@t> umn.edu Wed Feb 8 11:51:38 2006 From: meint002 <@t> umn.edu (meint002) Date: Wed Feb 8 11:51:49 2006 Subject: [Histonet] request feedback on benchtop tissue processors Message-ID: <200602081751.k18HpcSv018541@vanguard.software.umn.edu> I have been newly hired to do tissue research concerning muscular dystrophy and genetic cases. The lab wants to purchase a tissue processor and since the sample volume would be low, we are looking a benchtop model. So far I have read up about the following models; Leica TP1020, Sakura Tissue Tek VIP 150, TBS ATP-120 by Triangle Biomedical Sciences, and STP 120 spin tissue processors. Please let me know if you either have one of these models and what you like or dislike about it. I would appreciate all feedback you can provide. We are also looking into the purchase of a low volume stainer for immunohistochemical staining. I would appreciate any feedback on these models as well. Thanks very much. Joyce Meints Histologist University of Minnesota Paul and Sheila Wellstone Muscular Dystrophy Center MMC 206 420 Delaware St. SE Minneapolis, MN 55455-0392 e-mail: meint002@umn.edu lab phone: 612-626-4703 From rjbuesa <@t> yahoo.com Wed Feb 8 11:56:10 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Feb 8 11:56:18 2006 Subject: [Histonet] request feedback on benchtop tissue processors In-Reply-To: <200602081751.k18HpcSv018541@vanguard.software.umn.edu> Message-ID: <20060208175610.34431.qmail@web61213.mail.yahoo.com> Joyce: According with my experience Sakura tissue procesors are the best. By the way it does not matter if they are benchtop or "self standing", both have the same capacity. One model or the other will depend on your bench top availability. Ren? J. meint002 wrote: I have been newly hired to do tissue research concerning muscular dystrophy and genetic cases. The lab wants to purchase a tissue processor and since the sample volume would be low, we are looking a benchtop model. So far I have read up about the following models; Leica TP1020, Sakura Tissue Tek VIP 150, TBS ATP-120 by Triangle Biomedical Sciences, and STP 120 spin tissue processors. Please let me know if you either have one of these models and what you like or dislike about it. I would appreciate all feedback you can provide. We are also looking into the purchase of a low volume stainer for immunohistochemical staining. I would appreciate any feedback on these models as well. Thanks very much. Joyce Meints Histologist University of Minnesota Paul and Sheila Wellstone Muscular Dystrophy Center MMC 206 420 Delaware St. SE Minneapolis, MN 55455-0392 e-mail: meint002@umn.edu lab phone: 612-626-4703 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! From pruegg <@t> ihctech.net Wed Feb 8 12:10:01 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Wed Feb 8 12:10:11 2006 Subject: [Histonet] request feedback on benchtop tissue processors In-Reply-To: <20060208175610.34431.qmail@web61213.mail.yahoo.com> Message-ID: <200602081810.k18I9xDZ001624@chip.viawest.net> Joyce, You might also want to consider the microwave tissue processors, there are several out there, I have heard good things about the ?Miles system. As for autostainers. The Dako Autostainer is not small capacity as it will do 48 slides at once (you do not have to use it at capacity) for me it is the most cost effective because it is a completely open system which means you can shop around for reagents and not get stuck with the very expensive per packaged kits some are locked into. Also, for research purposes the DAKO is a lot more versatile as it is easy to use more exotic antibodies that may be of a species other than mouse or rabbit. My thoughts, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Wednesday, February 08, 2006 10:56 AM To: meint002; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] request feedback on benchtop tissue processors Joyce: According with my experience Sakura tissue procesors are the best. By the way it does not matter if they are benchtop or "self standing", both have the same capacity. One model or the other will depend on your bench top availability. Ren? J. meint002 wrote: I have been newly hired to do tissue research concerning muscular dystrophy and genetic cases. The lab wants to purchase a tissue processor and since the sample volume would be low, we are looking a benchtop model. So far I have read up about the following models; Leica TP1020, Sakura Tissue Tek VIP 150, TBS ATP-120 by Triangle Biomedical Sciences, and STP 120 spin tissue processors. Please let me know if you either have one of these models and what you like or dislike about it. I would appreciate all feedback you can provide. We are also looking into the purchase of a low volume stainer for immunohistochemical staining. I would appreciate any feedback on these models as well. Thanks very much. Joyce Meints Histologist University of Minnesota Paul and Sheila Wellstone Muscular Dystrophy Center MMC 206 420 Delaware St. SE Minneapolis, MN 55455-0392 e-mail: meint002@umn.edu lab phone: 612-626-4703 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Feb 8 12:34:23 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Feb 8 12:34:38 2006 Subject: Sorry reply was for everyone on floor model vs Re: [Histonet] request feedback on benchtop tissue processors In-Reply-To: <20060208175610.34431.qmail@web61213.mail.yahoo.com> References: <200602081751.k18HpcSv018541@vanguard.software.umn.edu> <20060208175610.34431.qmail@web61213.mail.yahoo.com> Message-ID: <6.0.0.22.1.20060208113310.01b4da28@gemini.msu.montana.edu> The joy of any floor model is easy access to rear of machine for repairs, etc - they roll around very nicely or put the bench model on a sturdy cart so you can access the back of instrument. At 10:56 AM 2/8/2006, you wrote: >Joyce: > According with my experience Sakura tissue procesors are the best. By > the way it does not matter if they are benchtop or "self standing", both > have the same capacity. One model or the other will depend on your bench > top availability. > Ren? J. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From pathrm35 <@t> adelphia.net Wed Feb 8 12:50:12 2006 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Wed Feb 8 12:50:21 2006 Subject: [Histonet] Alcohol Disposal Message-ID: <18782805.1139424612490.JavaMail.root@web24> Samuel, We have been using a chemical waste company to pick up our waste alcohol and xylene. They have proven to be unreliable and expensive so we sought another means. We are currently testing a CBG recycler for our alcohol and xylene waste. We have had very good results and have reduced our waste ten fold. Ron Martin ---- Samuel.Jones2@med.va.gov wrote: > > > Hello All: > > > > I've been asked by my peers to see how labs are collecting and disposing of > their waste alcohol. Thoughts please. Thanks > > > > Samuel E. Jones, MS, HT(ASCP)HTL, QIHC > > Supervisor, Anatomic Pathology > > VA North Texas Health Care System > > 4500 South Lancaster Road, 113 > > Dallas, Texas 75216 > > Phone: 214-857-0659 > > e-mail: samuel.jones2@med.va.gov > > > From nadams <@t> CapeCodHealth.org Wed Feb 8 13:22:37 2006 From: nadams <@t> CapeCodHealth.org (Adams, Nancy) Date: Wed Feb 8 13:23:11 2006 Subject: [Histonet] microwave with ventilation capabilities/vendors welcome Message-ID: <17A1862099540D458C8FE9380C2BC461C09330@fh2xmail.fhdomain1.capecodhealth.org> We are looking for a new microwave that can be ventilated to meet the new CAP question ANP.29430. We are using microwave outside of a hood and have existing ventilation to hook it in to. Thanks. Nancy Rutledge ************************************************************ This email and any files transmitted with it are confidential. And intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please contact the system administrator for Cape Cod Healthcare. Administrator@CapeCodHealth.org ************************************************************ From gpbnas <@t> yahoo.es Wed Feb 8 14:04:48 2006 From: gpbnas <@t> yahoo.es (Guillermo Palao) Date: Wed Feb 8 14:04:59 2006 Subject: [Histonet] Weak hematoxylin counterstaining in immunohistochemistry Message-ID: <20060208200448.68755.qmail@web26209.mail.ukl.yahoo.com> Dear All, I am quite new to the histological techniques, but the fact is that I have been counterstaining with Harris Hematoxylin immunohistochemistry frozen samples fixed with acetone without problems until now. With different antibodies but similarly prepared slides counterstaining of nuclei is very weak, only their outer ring being clearly stained. Why is this happpening? Any help on how to resolve this issue is really welcomed. Guillermo Palao, MD. Ph.D. Laboratorio de Reumatolog?a Centro de Investigaci?n Hospital 12 de Octubre Avda de C?rdoba s/n Madrid 28041 Spain --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com From Eric <@t> ategra.com Wed Feb 8 13:58:16 2006 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Wed Feb 8 14:41:32 2006 Subject: [Histonet] Asking for your help ... Message-ID: Hi Histonetters, I just wanted to give you a warning about Pam Barker/Relia Inc, and ask for your help. Here at Ategra, we used to have an employee named Pam Barker. She resigned from Ategra last year (Jan 20 2005) and started her own company named Relia LLC. Since she resigned we here at Ategra were alarmed to find that Pam Barker has contacted Ategra clients and candidates in violation of her non-compete agreement. The court has agreed and has issued an Injunction (like a restraining order) against Pam Barker/Relia LLC. This Injunction (like a restraining order) specifically prohibits Pam Barker from contacting Ategra clients/candidates (including prospective clients/candidates) From Jan 05 2006 until Aug 8th 2006 (if not later) Because I strive to keep Ategra's contact lists strictly confidential, I would like to ask a favor of you: The favor I ask is this : if you have been contacted by Pam Barker OR Relia LLC if you could PLEASE LET ME KNOW ? Your name will be kept confidential and no other involvement from you or your firm is necessary. All I need to know is the appx date she has contacted you. Thank You in Advance For Your Help Eric Dye, PS: If you have any questions about this, call me at 800-466-9919 ext 223 PSS: If you need to see a copy of the Injunction (restraining order) against Pam Barker, click here: http://www.ategra.com/injunction.html PSSS: I have openings for HistoTechs throughout the US. If you are thinking of a making career move, feel free to call me. ---------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd Winter Park, FL 32792-6721 VOICE: 800-466-9919 ext 223 FAX: 407-671-6075 EMAIL: Eric@ategra.com ---------------------------------------------------- From LuckG <@t> empirehealth.org Wed Feb 8 14:42:13 2006 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Wed Feb 8 14:42:24 2006 Subject: [Histonet] microwave with ventilation capabilities/vendors we lcome Message-ID: Nancy, et.al., I answered this question as "N/A" (not applicable) as the reagents we use in the microwave are only "non-hazardous" (e.g. aqueous solutions, most biological stains, no pathogens or carcinogens. More specifically we are not processing any tissues or using common tissue processing agents. See the comment section for this CAP question. Does this seem to be an appropriate interpretation of the intent of this question? Thanks, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org www.deaconessmedicalcenter.org -----Original Message----- From: Adams, Nancy [mailto:nadams@CapeCodHealth.org] Sent: Wednesday, February 08, 2006 11:23 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] microwave with ventilation capabilities/vendors welcome We are looking for a new microwave that can be ventilated to meet the new CAP question ANP.29430. We are using microwave outside of a hood and have existing ventilation to hook it in to. Thanks. Nancy Rutledge ************************************************************ This email and any files transmitted with it are confidential. And intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please contact the system administrator for Cape Cod Healthcare. Administrator@CapeCodHealth.org ************************************************************ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dellav <@t> musc.edu Wed Feb 8 15:01:29 2006 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Wed Feb 8 15:02:07 2006 Subject: [Histonet] microwave with ventilation capabilities/vendors welcome Message-ID: Greg's interpretation is correct. if you are not heating solutions that will lead to hazardous vapors, you have met the standard. Vinnie Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> "Luck, Greg D." 02/08/06 03:42PM >>> Nancy, et.al., I answered this question as "N/A" (not applicable) as the reagents we use in the microwave are only "non-hazardous" (e.g. aqueous solutions, most biological stains, no pathogens or carcinogens. More specifically we are not processing any tissues or using common tissue processing agents. See the comment section for this CAP question. Does this seem to be an appropriate interpretation of the intent of this question? Thanks, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org www.deaconessmedicalcenter.org -----Original Message----- From: Adams, Nancy [mailto:nadams@CapeCodHealth.org] Sent: Wednesday, February 08, 2006 11:23 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] microwave with ventilation capabilities/vendors welcome We are looking for a new microwave that can be ventilated to meet the new CAP question ANP.29430. We are using microwave outside of a hood and have existing ventilation to hook it in to. Thanks. Nancy Rutledge ************************************************************ This email and any files transmitted with it are confidential. And intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please contact the system administrator for Cape Cod Healthcare. Administrator@CapeCodHealth.org ************************************************************ _______________________________________________ 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 From mike.voor <@t> louisville.edu Wed Feb 8 18:47:30 2006 From: mike.voor <@t> louisville.edu (Michael J Voor) Date: Wed Feb 8 18:50:49 2006 Subject: [Histonet] plastic embedded bone sections Message-ID: Dear Histonet, We are trying to section PMMA (Technovit 9100 New) embedded undecalcified bone samples (rat tibias). We plan to use a Microm sliding microtome. We need advice on exactly what blade to use and what blade angle would be best to create sections on the order of 7-10 microns. Thanks for any help. Michael J. Voor, Ph.D. Orthopaedic Bioengineering Laboratory Director of Research Department of Orthopaedic Surgery University of Louisville Louisville, KY 40292 (502) 852-7673 FAX: (502)852-7227 e-mail: mike.voor@louisville.edu From louise.renton <@t> gmail.com Thu Feb 9 01:13:37 2006 From: louise.renton <@t> gmail.com (louise renton) Date: Thu Feb 9 01:13:45 2006 Subject: [Histonet] plastic embedded bone sections In-Reply-To: References: Message-ID: We routinely cut human and non-human primate bones embedded in 9100 new on an automated Leica SM2500 microtome ( not familiar with the microm model - is it automated?). We use a tungsten carbide tipped knife to cut 6mu sections. I best regards On 2/9/06, Michael J Voor wrote: > Dear Histonet, > > We are trying to section PMMA (Technovit 9100 New) embedded > undecalcified bone samples (rat tibias). We plan to use a Microm > sliding microtome. We need advice on exactly what blade to use and what > blade angle would be best to create sections on the order of 7-10 > microns. Thanks for any help. > > > Michael J. Voor, Ph.D. > Orthopaedic Bioengineering Laboratory > Director of Research > Department of Orthopaedic Surgery > University of Louisville > Louisville, KY 40292 > (502) 852-7673 FAX: (502)852-7227 > e-mail: mike.voor@louisville.edu > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "Does a conceited cowboy, only ride on pompous grass?. From ree3 <@t> leicester.ac.uk Thu Feb 9 04:09:39 2006 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Thu Feb 9 04:09:49 2006 Subject: [Histonet] mouse ovaries Message-ID: Anyone know where I can find out about the histology of mouse ovaries??? Many thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER...U.K...... From BMolinari <@t> heart.thi.tmc.edu Thu Feb 9 05:29:44 2006 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Thu Feb 9 05:36:16 2006 Subject: [Histonet] Alcohol Disposal Message-ID: We collect all alcohols that are above a 70% solution. The waste is picked up by the hospital and we pay a fee for disposal. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Samuel.Jones2@med.va.gov Sent: Wednesday, February 08, 2006 9:08 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Alcohol Disposal Hello All: I've been asked by my peers to see how labs are collecting and disposing of their waste alcohol. Thoughts please. Thanks Samuel E. Jones, MS, HT(ASCP)HTL, QIHC Supervisor, Anatomic Pathology VA North Texas Health Care System 4500 South Lancaster Road, 113 Dallas, Texas 75216 Phone: 214-857-0659 e-mail: samuel.jones2@med.va.gov From pmcardle <@t> ebsciences.com Thu Feb 9 06:46:10 2006 From: pmcardle <@t> ebsciences.com (Phil McArdle) Date: Thu Feb 9 06:46:46 2006 Subject: [Histonet] microwave with ventilation capabilities/vendors welcome In-Reply-To: <17A1862099540D458C8FE9380C2BC461C09330@fh2xmail.fhdomain1.capecodhealth.org> References: <17A1862099540D458C8FE9380C2BC461C09330@fh2xmail.fhdomain1.capecodhealth.org> Message-ID: <43EB3992.9050900@ebsciences.com> Hello: Energy Beam Sciences, a pioneer in microwave-assisted staining and tissue processing, has both general-purpose ("staining") and processing laboratory microwaves. All our microwaves feature powered fume extraction systems with vent-system interlock, which shuts the microwave down in the event of vent obstruction or failure. Currently, our H2200 model is the lowest-priced vented laboratory microwave available. We also offer a wide range of microwave- and histology-related accessories, including an affordable microwave leakage detector (model H2501). We are always happy to provide quotations, and in the case of our H2850, demonstration units are available (and encouraged). We invite you to visit our website: http://www.ebsstore.com/control/category/~category_id=C Best regards, Phil McArdle -- Phil McArdle Microwave Product Manager Energy Beam Sciences, Inc. Tel: 800.992.9037 x 341 Fax: 860.653.0422 PMcardle@ebsciences.com www.ebsciences.com "ADDING BRILLIANCE TO YOUR VISION" Adams, Nancy wrote: > > > We are looking for a new microwave that can be ventilated to > meet the new CAP question ANP.29430. We are using microwave outside of > a hood and have existing ventilation to hook it in to. > > Thanks. > > Nancy Rutledge > > > > ************************************************************ > This email and any files transmitted with it are confidential. And intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please contact the system administrator for Cape Cod Healthcare. > Administrator@CapeCodHealth.org > ************************************************************ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From kmerriam2003 <@t> yahoo.com Thu Feb 9 06:53:24 2006 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Thu Feb 9 06:53:32 2006 Subject: [Histonet] Slide scanner for Quantum Dot Fluorescence Message-ID: <20060209125324.62946.qmail@web50309.mail.yahoo.com> Hello All, I am looking for a slide scanner (captures images of entire tissue sections on slides) that has fluorescent capabilities. We currently have an Aperio slide scanner for our routine light microscopy slides, we are very happy with it, but we will be branching out into Quantum Dots and would like a scanner that can do the same thing for fluorescent-stained slides. Does anyone have any ideas or recommendations? Thanks, Kim Kim Merriam Novartis Cambridge, MA --------------------------------- Yahoo! Mail - Helps protect you from nasty viruses. From sluhisto <@t> yahoo.com Thu Feb 9 07:19:32 2006 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Thu Feb 9 07:19:43 2006 Subject: [Histonet] Asking for your help ... In-Reply-To: Message-ID: <20060209131932.7826.qmail@web51006.mail.yahoo.com> Eric: I do not believe that this is the place for this type of post. As I indicated to you in a private post, I personally do not believe that we here on the histonet can be considered "clients" simply because you have posted here or even if one of us has asked about a position. I feel that the only people that you can consider "clients" are those who have signed up for your services either as an employer or employee that you have placed. I find your post offensive and unprofessional. You have every right to contact your true clients and elicit their support in your endeavors but I feel that posting here is inappropriate. Susan Eric Dye wrote: Hi Histonetters, I just wanted to give you a warning about Pam Barker/Relia Inc, and ask for your help. Here at Ategra, we used to have an employee named Pam Barker. She resigned from Ategra last year (Jan 20 2005) and started her own company named Relia LLC. Since she resigned we here at Ategra were alarmed to find that Pam Barker has contacted Ategra clients and candidates in violation of her non-compete agreement. The court has agreed and has issued an Injunction (like a restraining order) against Pam Barker/Relia LLC. This Injunction (like a restraining order) specifically prohibits Pam Barker from contacting Ategra clients/candidates (including prospective clients/candidates) From Jan 05 2006 until Aug 8th 2006 (if not later) Because I strive to keep Ategra's contact lists strictly confidential, I would like to ask a favor of you: The favor I ask is this : if you have been contacted by Pam Barker OR Relia LLC if you could PLEASE LET ME KNOW ? Your name will be kept confidential and no other involvement from you or your firm is necessary. All I need to know is the appx date she has contacted you. Thank You in Advance For Your Help Eric Dye, PS: If you have any questions about this, call me at 800-466-9919 ext 223 PSS: If you need to see a copy of the Injunction (restraining order) against Pam Barker, click here: http://www.ategra.com/injunction.html PSSS: I have openings for HistoTechs throughout the US. If you are thinking of a making career move, feel free to call me. ---------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd Winter Park, FL 32792-6721 VOICE: 800-466-9919 ext 223 FAX: 407-671-6075 EMAIL: Eric@ategra.com ---------------------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! From DDDeltour <@t> mar.med.navy.mil Thu Feb 9 07:48:13 2006 From: DDDeltour <@t> mar.med.navy.mil (DDDeltour@mar.med.navy.mil) Date: Thu Feb 9 07:48:51 2006 Subject: [Histonet] Asking for your help ... Message-ID: <3F500F8B416C554EBB21FF16642F72E959CDBE@marxchg03.mar.med.navy.mil> Susan, I have to agree with your assessment. A private email would have been sufficient. I am not a client of anyone. Both Eric and Pam have posted jobs on here and have emailed with jobs. I am not going to "sign" with only one "headhunter" anyway. It would be ridiculous to limit myself to one set of job opportunities. I consider myself a free agent and open to the highest bidder :) Douglas D. Deltour HT(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histology SLU Sent: Thursday, February 09, 2006 8:20 AM To: Eric Dye; Histonetters Subject: Re: [Histonet] Asking for your help ... Eric: I do not believe that this is the place for this type of post. As I indicated to you in a private post, I personally do not believe that we here on the histonet can be considered "clients" simply because you have posted here or even if one of us has asked about a position. I feel that the only people that you can consider "clients" are those who have signed up for your services either as an employer or employee that you have placed. I find your post offensive and unprofessional. You have every right to contact your true clients and elicit their support in your endeavors but I feel that posting here is inappropriate. Susan Eric Dye wrote: Hi Histonetters, I just wanted to give you a warning about Pam Barker/Relia Inc, and ask for your help. Here at Ategra, we used to have an employee named Pam Barker. She resigned from Ategra last year (Jan 20 2005) and started her own company named Relia LLC. Since she resigned we here at Ategra were alarmed to find that Pam Barker has contacted Ategra clients and candidates in violation of her non-compete agreement. The court has agreed and has issued an Injunction (like a restraining order) against Pam Barker/Relia LLC. This Injunction (like a restraining order) specifically prohibits Pam Barker from contacting Ategra clients/candidates (including prospective clients/candidates) From Jan 05 2006 until Aug 8th 2006 (if not later) Because I strive to keep Ategra's contact lists strictly confidential, I would like to ask a favor of you: The favor I ask is this : if you have been contacted by Pam Barker OR Relia LLC if you could PLEASE LET ME KNOW ? Your name will be kept confidential and no other involvement from you or your firm is necessary. All I need to know is the appx date she has contacted you. Thank You in Advance For Your Help Eric Dye, PS: If you have any questions about this, call me at 800-466-9919 ext 223 PSS: If you need to see a copy of the Injunction (restraining order) against Pam Barker, click here: http://www.ategra.com/injunction.html PSSS: I have openings for HistoTechs throughout the US. If you are thinking of a making career move, feel free to call me. ---------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd Winter Park, FL 32792-6721 VOICE: 800-466-9919 ext 223 FAX: 407-671-6075 EMAIL: Eric@ategra.com ---------------------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bob-meyer <@t> northwestern.edu Thu Feb 9 07:52:40 2006 From: bob-meyer <@t> northwestern.edu (bob-meyer@northwestern.edu) Date: Thu Feb 9 07:52:48 2006 Subject: [Histonet] CISH Message-ID: <20060209135240.86B2535C55@casbah.it.northwestern.edu> The steps for performing CISH are not simple. I suggest using the GenPoint kit from DAKO (K0620). Other companies have kits as well, and most companies put the product insert on their web page. That will give you some idea of the steps and complexity involved. Bob Meyer Northwestern University ==============Original message text=============== On Tue, 07 Feb 2006 6:47:46 pm +0000 "Muhammad Tahseen" wrote: Hi All, I was wondering if anyone can let me know the simple steps involved in performing CISH . Thanks in advance, Muhammad Tahseen, MLT(Punjab Medical Faculty) MT(JIMTEF)Japan Histology Supervisor SKMCH & RC Pakistan. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ===========End of original message text=========== From SARAH.REEVES <@t> ekht.nhs.uk Thu Feb 9 08:14:56 2006 From: SARAH.REEVES <@t> ekht.nhs.uk (SARAH REEVES) Date: Thu Feb 9 08:15:58 2006 Subject: [Histonet] Automated ABDPAS Message-ID: Does anyone automate their alcian blue diastase periodic acid schiffs? Do you do the whole or part of the stain on the stainer? Are your solutions fresh or reused? Any advice would be appreciated as we are looking to automate ABDPAS and giemsa for gastric biopsies. Sarah >>> histonet-request@lists.utsouthwestern.edu 02/08/06 6:01 pm >>> 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-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@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. re: Technovit 7100 resin removal (Carl Hobbs) 2. RE: Celestine blue B (Sharon Allen) 3. argyrophil and argentaffin (Till, Renee) 4. re: argyrophil and argentaffin (bhewlett@cogeco.ca) 5. re: argyrophil and argentaffin (bhewlett@cogeco.ca) 6. Alcohol Disposal (Samuel.Jones2@med.va.gov) 7. RE: Celestine blue B (Rittman, Barry R) 8. Fwd: Re: [Histonet] Alcohol Disposal (Atoska S. Gentry) 9. backup cryostat (Steven Coakley) 10. infrared vs uv in sequencing (Malcolm McCallum) 11. Alcohol Disposal--a few choices (Cheryl) 12. request feedback on benchtop tissue processors (meint002) 13. Re: request feedback on benchtop tissue processors (Rene J Buesa) ---------------------------------------------------------------------- Message: 1 Date: Wed, 8 Feb 2006 14:07:58 -0000 From: "Carl Hobbs" Subject: [Histonet] re: Technovit 7100 resin removal To: Message-ID: <001201c62cb9$10d89880$112b5c9f@Carlos> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original In all my years, I have never been able to remove the resin ( of ANY GMA-based resin) and yet still retain the section fixed to a slide, sigh! I bought the same Technovit resin for immunohistochemical applications yet have never found it to be a viable alternative to pwax sections. Methyl methac is the only resin that can be removed, IMHO, still retaining the section on the slide. Carl ------------------------------ Message: 2 Date: Wed, 8 Feb 2006 08:34:14 -0600 From: Sharon Allen Subject: RE: [Histonet] Celestine blue B To: 'Julien Lambrey de Souza' , "Histonet (E-mail)" Message-ID: <36FE435D6D2F1D489174B22362A961B66B8313@hscxntmx0006.hsc.mb.ca> Content-Type: text/plain; charset="us-ascii" Hi, I started using Celestine Blue in our method for Gomori's Trichrome & it made a great improvement. I'm not sure why! Put the slides in Celestine Blue for 2 min. before the Mayer's Haematoxylin. Recipe is: Ferric ammonium sulfate...........5.0 gm Dissolve in distilled water.......100 ml. Celestine blue....................0.5 gm Boil for 3 min. to dissolve, filter, when cooled add- Glycerin..........................14 ml Stable for at least 6 months (Carleton's Histological Techniques 4th edition P.133) Hope this helps. Sharon sallen@hsc.mb.ca -----Original Message----- This e-mail and/or any documents in this transmission is intended for the address(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. ------------------------------ Message: 3 Date: Wed, 8 Feb 2006 08:43:26 -0600 From: "Till, Renee" Subject: [Histonet] argyrophil and argentaffin To: histonet@lists.utsouthwestern.edu Message-ID: <11F927674DEBDC43B960809A7403C5D232866C@MAILPED.ad.uams.edu> Content-Type: text/plain; charset=us-ascii I had heard that if you are doing a Grimelius for argyrophil cells, that an argentaffin stain should also be performed to be able to differentiate between the two cells. Is this true for any argyrophil stain? Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 ================================================================================================ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ================================================================================================ ------------------------------ Message: 4 Date: Wed, 08 Feb 2006 10:13:41 -0500 From: bhewlett@cogeco.ca Subject: re: [Histonet] argyrophil and argentaffin To: "Till,Renee" , histonet@lists.utsouthwestern.edu Message-ID: <43ea0aa5.322.b62.22262@cogeco.ca> > Renee, All argentaffin cells are also argyrophil by definition. Therefore, to distinguish the true argyrophil population, one should perform both stains and subtract the argentaffin population from the argyrophil. Bryan > I had heard that if you are doing a Grimelius for argyrophil cells, that > an argentaffin stain should also be performed to be able to > differentiate between the two cells. Is this true for any argyrophil > stain? > > > > Renee' Till, HT > > > > Arkansas Childrens Nutrition Center > > 1120 Marshall > > Little Rock, AR > > 72202 > > > > > > > =============================================================================================== > Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended > recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or > distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and > destroy all copies of the original message. > ===============================================================================================_______________________ > ________________________ Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Wed, 08 Feb 2006 10:15:24 -0500 From: bhewlett@cogeco.ca Subject: re: [Histonet] argyrophil and argentaffin To: "Till,Renee" , histonet@lists.utsouthwestern.edu Message-ID: <43ea0b0c.40.65c1.3974@cogeco.ca> > Renee, All argentaffin cells are also argyrophil by definition. Therefore, to distinguish the true argyrophil population, one should perform both stains and subtract the argentaffin population from the argyrophil. Bryan > I had heard that if you are doing a Grimelius for argyrophil cells, that > an argentaffin stain should also be performed to be able to > differentiate between the two cells. Is this true for any argyrophil > stain? > > > > Renee' Till, HT > > > > Arkansas Childrens Nutrition Center > > 1120 Marshall > > Little Rock, AR > > 72202 > > > > > > > =============================================================================================== > Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended > recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or > distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and > destroy all copies of the original message. > ===============================================================================================_______________________ > ________________________ Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Wed, 8 Feb 2006 09:08:12 -0600 From: Samuel.Jones2@med.va.gov Subject: [Histonet] Alcohol Disposal To: histonet@lists.utsouthwestern.edu Message-ID: <476BAB689BEA3E4DBE6F8D5A90CAC2CF056FC7DA@vhantxexc1.v17.med.va.gov> Content-Type: text/plain; charset="us-ascii" Hello All: I've been asked by my peers to see how labs are collecting and disposing of their waste alcohol. Thoughts please. Thanks Samuel E. Jones, MS, HT(ASCP)HTL, QIHC Supervisor, Anatomic Pathology VA North Texas Health Care System 4500 South Lancaster Road, 113 Dallas, Texas 75216 Phone: 214-857-0659 e-mail: samuel.jones2@med.va.gov ------------------------------ Message: 7 Date: Wed, 8 Feb 2006 09:42:40 -0600 From: "Rittman, Barry R" Subject: RE: [Histonet] Celestine blue B To: Message-ID: Content-Type: text/plain; charset="us-ascii" We used to use this formula for general staining with Celestine blue but came across a formula in the literature many years ago that gave us a much more intense stain and also lasted a much longer time. Sorry but cannot remember the reference. >From memory so hope that this is correct. 0.5 gms Celestine blue powder into a glass beaker. Under the fume hood add 0.5 ml of concentrated sulfuric acid and mix with a glass rod into a paste. During this time the mixture bubbles and gives off fumes hence the fume hood. Add 100 ml of 5% aqueous iron alum containing 14 ml of glycerin. Mix well and filter. This solution stains very rapidly to start with and staining time decreases a little after that so you need to test it on sections for half a minute when it is first made. A good way to test whether Celestine blue stains are past their useful staining life is to swirl the solution around the bottle or flask and if the solution fails to stick somewhat to the glass then need to make up a fresh batch. Filter each time used. We really liked this modification as the nuclear stain when using van Gieson stain as it is not differentiated by the picric acid. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sharon Allen Sent: Wednesday, February 08, 2006 8:34 AM To: 'Julien Lambrey de Souza'; Histonet (E-mail) Subject: RE: [Histonet] Celestine blue B Hi, I started using Celestine Blue in our method for Gomori's Trichrome & it made a great improvement. I'm not sure why! Put the slides in Celestine Blue for 2 min. before the Mayer's Haematoxylin. Recipe is: Ferric ammonium sulfate...........5.0 gm Dissolve in distilled water.......100 ml. Celestine blue....................0.5 gm Boil for 3 min. to dissolve, filter, when cooled add- Glycerin..........................14 ml Stable for at least 6 months (Carleton's Histological Techniques 4th edition P.133) Hope this helps. Sharon sallen@hsc.mb.ca -----Original Message----- This e-mail and/or any documents in this transmission is intended for the address(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. ------------------------------ Message: 8 Date: Wed, 08 Feb 2006 09:47:36 -0600 From: "Atoska S. Gentry" Subject: Fwd: Re: [Histonet] Alcohol Disposal To: Histonet Message-ID: <6.0.1.1.0.20060208094734.01a81e48@mailhost.vetmed.auburn.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed >Date: Wed, 08 Feb 2006 09:47:25 -0600 >To: Samuel.Jones2@med.va.gov >From: "Atoska S. Gentry" >Subject: Re: [Histonet] Alcohol Disposal > > >Hello, our campus' Risk Management Department picks up our waste Alcohol >along with other chemical waste. I'm not sure of their means of disposal. >Best wishes. Atoska > > >At 09:08 AM 2/8/2006, you wrote: >> >> >>Hello All: >> >> >> >>I've been asked by my peers to see how labs are collecting and disposing of >>their waste alcohol. Thoughts please. Thanks >> >> >> >>Samuel E. Jones, MS, HT(ASCP)HTL, QIHC >> >>Supervisor, Anatomic Pathology >> >>VA North Texas Health Care System >> >>4500 South Lancaster Road, 113 >> >>Dallas, Texas 75216 >> >>Phone: 214-857-0659 >> >>e-mail: samuel.jones2@med.va.gov >> >> >> >>_______________________________________________dd 0 >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Wed, 8 Feb 2006 08:18:09 -0800 (PST) From: Steven Coakley Subject: [Histonet] backup cryostat To: Histonet@lists.utsouthwestern.edu Message-ID: <20060208161809.48345.qmail@web90208.mail.scd.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Our company is looking for a backup cryostat. Steve --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! ------------------------------ Message: 10 Date: Wed, 8 Feb 2006 10:39:25 -0600 From: "Malcolm McCallum" Subject: [Histonet] infrared vs uv in sequencing To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" anyone familiar with the advantages/disadvantages? Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Till, Renee Sent: Wed 2/8/2006 8:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] argyrophil and argentaffin I had heard that if you are doing a Grimelius for argyrophil cells, that an argentaffin stain should also be performed to be able to differentiate between the two cells. Is this true for any argyrophil stain? Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 ================================================================================================ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ================================================================================================ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Wed, 8 Feb 2006 08:52:15 -0800 (PST) From: Cheryl Subject: [Histonet] Alcohol Disposal--a few choices To: samuel.jones2@med.va.gov, histonet@lists.utsouthwestern.edu Message-ID: <20060208165216.21889.qmail@web50902.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi Samuel- This kind of issue usually starts with the water department in your district. Some places have enough water going through their system that this kind of waste can go down the drain. It is considered safe and cost effective ($0!!) because of the dilution factors. Hospitals use HUGE quantities of water so they might even make an exception if it's not in the base policies. Before you contact your water district, talk with your risk management and environmental services people--they may already know or might want to talk to the water people for you. If you end up not being able to dump it, (and the water company can usually answer you within a few days) you may want to consider recycling. BR and one other company (??) can do a cost analysis with just a few questions to determine if this won't SAVE your department a buncha bucks (including your people's time in the equasion.) These systems are SO effective the end result is sometimes better than the original solutions you got from your suppliers. You can do just alcohols or both Alc. and Xylenes. These systems are infinitely more useable and safer than they were 20 years ago. The most expensive options are usually waste haulers and they will charge you more for certain contaminants. Xylene, biologicals, etc. So when you get a quote from a waste hauler or incinerator, ask if they are going to recycle it themselves (should get you a lower price) or burn it, and any additional cost for other cross-contaminants from your stain line or processors. Being a tree-hugger from way back, I always chose the recycle method when it is fiscally responsible (and it usually is). Hope this helps! Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one tech at a time. admin@fullstaff.org 281.852.9457 o 281.883.7704 c "Atoska S. Gentry" wrote: >Date: Wed, 08 Feb 2006 09:47:25 -0600 >To: Samuel.Jones2@med.va.gov >From: "Atoska S. Gentry" >Subject: Re: [Histonet] Alcohol Disposal > > >Hello, our campus' Risk Management Department picks up our waste Alcohol >along with other chemical waste. I'm not sure of their means of disposal. >Best wishes. Atoska > > >At 09:08 AM 2/8/2006, you wrote: >> >> >>Hello All: >> >> >> >>I've been asked by my peers to see how labs are collecting and disposing of >>their waste alcohol. Thoughts please. Thanks >> >> >> >>Samuel E. Jones, MS, HT(ASCP)HTL, QIHC >> >>Supervisor, Anatomic Pathology >> >>VA North Texas Health Care System >> >>4500 South Lancaster Road, 113 >> >>Dallas, Texas 75216 >> >>Phone: 214-857-0659 >> >>e-mail: samuel.jones2@med.va.gov >> >> >> >>_______________________________________________ >>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 ------------------------------ Message: 12 Date: Wed, 08 Feb 2006 11:51:38 CST From: meint002 Subject: [Histonet] request feedback on benchtop tissue processors To: histonet@lists.utsouthwestern.edu Message-ID: <200602081751.k18HpcSv018541@vanguard.software.umn.edu> Content-Type: TEXT/plain; CHARSET=US-ASCII I have been newly hired to do tissue research concerning muscular dystrophy and genetic cases. The lab wants to purchase a tissue processor and since the sample volume would be low, we are looking a benchtop model. So far I have read up about the following models; Leica TP1020, Sakura Tissue Tek VIP 150, TBS ATP-120 by Triangle Biomedical Sciences, and STP 120 spin tissue processors. Please let me know if you either have one of these models and what you like or dislike about it. I would appreciate all feedback you can provide. We are also looking into the purchase of a low volume stainer for immunohistochemical staining. I would appreciate any feedback on these models as well. Thanks very much. Joyce Meints Histologist University of Minnesota Paul and Sheila Wellstone Muscular Dystrophy Center MMC 206 420 Delaware St. SE Minneapolis, MN 55455-0392 e-mail: meint002@umn.edu lab phone: 612-626-4703 ------------------------------ Message: 13 Date: Wed, 8 Feb 2006 09:56:10 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] request feedback on benchtop tissue processors To: meint002 , histonet@lists.utsouthwestern.edu Message-ID: <20060208175610.34431.qmail@web61213.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Joyce: According with my experience Sakura tissue procesors are the best. By the way it does not matter if they are benchtop or "self standing", both have the same capacity. One model or the other will depend on your bench top availability. Ren* J. meint002 wrote: I have been newly hired to do tissue research concerning muscular dystrophy and genetic cases. The lab wants to purchase a tissue processor and since the sample volume would be low, we are looking a benchtop model. So far I have read up about the following models; Leica TP1020, Sakura Tissue Tek VIP 150, TBS ATP-120 by Triangle Biomedical Sciences, and STP 120 spin tissue processors. Please let me know if you either have one of these models and what you like or dislike about it. I would appreciate all feedback you can provide. We are also looking into the purchase of a low volume stainer for immunohistochemical staining. I would appreciate any feedback on these models as well. Thanks very much. Joyce Meints Histologist University of Minnesota Paul and Sheila Wellstone Muscular Dystrophy Center MMC 206 420 Delaware St. SE Minneapolis, MN 55455-0392 e-mail: meint002@umn.edu lab phone: 612-626-4703 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 27, Issue 12 **************************************** ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of East Kent Hospitals NHS Trust. If you are not the intended recipient, be advised that you have received this email in error and that any use, dissemination, forwarding, printing or copying of this email is strictly prohibited. If you have received this email in error please notify the system manager at the following email address: root.postmaster@ekht.nhs.uk www.kentandmedway.nhs.uk This footnote also confirms that although this email message has been swept by MIMEsweeper for the presence of computer viruses, it is strongly recommended that you carry out your own virus scan of this message and any attachments. www.mimesweeper.com ********************************************************************** From brian.chelack <@t> usask.ca Thu Feb 9 08:27:36 2006 From: brian.chelack <@t> usask.ca (Brian Chelack) Date: Thu Feb 9 08:29:27 2006 Subject: [Histonet] Microwave processors Message-ID: <000301c62d84$f998ee00$4f13e980@PDS11> Can any one provide me with information regarding microwave tissue processors? What is the quality of the sections derived from microwave processors compared to regular processors? Are there any reliability issues? Do you need to trim in very thin tissues? Anything else? Thanks Brian Chelack Prairie Diagnostic Services 2604-52 Campus Drive Saskatoon SK S7N 5B4 306-966-7241 From dusko.trajkovic <@t> pfizer.com Thu Feb 9 08:31:03 2006 From: dusko.trajkovic <@t> pfizer.com (Trajkovic, Dusko) Date: Thu Feb 9 08:31:13 2006 Subject: [Histonet] Slide scanner for Quantum Dot Fluorescence Message-ID: <3AD0BD3142459B4E9B12CBEAFF2B89B2D89759@lajamrexm01.amer.pfizer.com> Try contacting Ventana Medical Systems. 1-800-227-2155 Dusko Trajkovic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Merriam Sent: Thursday, February 09, 2006 4:53 AM To: Histonet Subject: [Histonet] Slide scanner for Quantum Dot Fluorescence Hello All, I am looking for a slide scanner (captures images of entire tissue sections on slides) that has fluorescent capabilities. We currently have an Aperio slide scanner for our routine light microscopy slides, we are very happy with it, but we will be branching out into Quantum Dots and would like a scanner that can do the same thing for fluorescent-stained slides. Does anyone have any ideas or recommendations? Thanks, Kim Kim Merriam Novartis Cambridge, MA --------------------------------- Yahoo! Mail - Helps protect you from nasty viruses. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------------------- LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From mucram11 <@t> comcast.net Thu Feb 9 08:41:33 2006 From: mucram11 <@t> comcast.net (Pam Marcum) Date: Thu Feb 9 08:41:46 2006 Subject: [Histonet] Asking for your help ... Message-ID: <020920061441.14759.43EB549D00053D47000039A72200734364CECE030E9D0C9A03@comcast.net> Thank you Susan. Well said and I agree. This is a private business matter between two companies and thier signed clients not all of HistoNet. While we appreciate knowing where the jobs we are not clients because we see and read the ads placed here. Pam Marcum -------------- Original message ---------------------- From: Histology SLU > Eric: > > I do not believe that this is the place for this type of post. As I indicated > to you in a private post, I personally do not believe that we here on the > histonet can be considered "clients" simply because you have posted here or even > if one of us has asked about a position. I feel that the only people that you > can consider "clients" are those who have signed up for your services either as > an employer or employee that you have placed. I find your post offensive and > unprofessional. You have every right to contact your true clients and elicit > their support in your endeavors but I feel that posting here is inappropriate. > > Susan > > Eric Dye wrote: > Hi Histonetters, I just wanted to give you a warning about Pam Barker/Relia > Inc, and ask for your help. > > Here at Ategra, we used to have an employee named Pam Barker. She resigned from > Ategra last year (Jan 20 2005) and started her own company named Relia LLC. > > Since she resigned we here at Ategra were alarmed to find that Pam Barker has > contacted Ategra clients and candidates in violation of her non-compete > agreement. > > The court has agreed and has issued an Injunction (like a restraining order) > against Pam Barker/Relia LLC. This Injunction (like a restraining order) > specifically prohibits Pam Barker from contacting Ategra clients/candidates > (including prospective clients/candidates) From Jan 05 2006 until Aug 8th 2006 > (if not later) > > Because I strive to keep Ategra's contact lists strictly confidential, I would > like to ask a favor of you: > > The favor I ask is this : if you have been contacted by Pam Barker OR Relia LLC > if you could PLEASE LET ME KNOW ? > > Your name will be kept confidential and no other involvement from you or your > firm is necessary. All I need to know is the appx date she has contacted you. > > Thank You in Advance For Your Help > > Eric Dye, > > PS: If you have any questions about this, call me at 800-466-9919 ext 223 > > PSS: If you need to see a copy of the Injunction (restraining order) against Pam > Barker, click here: http://www.ategra.com/injunction.html > > PSSS: I have openings for HistoTechs throughout the US. If you are thinking of a > making career move, feel free to call me. > > ---------------------------------------------------- > Ategra Systems Inc > Specialists in Permanent & Contract Staffing > 7085 University Blvd > Winter Park, FL 32792-6721 > > VOICE: 800-466-9919 ext 223 > FAX: 407-671-6075 > EMAIL: Eric@ategra.com > > ---------------------------------------------------- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > --------------------------------- > Relax. Yahoo! Mail virus scanning helps detect nasty viruses! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From HornHV <@t> archildrens.org Thu Feb 9 08:47:36 2006 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Thu Feb 9 08:47:58 2006 Subject: [Histonet] Asking for your help ... Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDF96@EMAIL.archildrens.org> You are absolutely correct Susan. This post offended me and this list cannot be considered Eric's client. Hazel -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histology SLU Sent: Thursday, February 09, 2006 7:20 AM To: Eric Dye; Histonetters Subject: Re: [Histonet] Asking for your help ... Eric: I do not believe that this is the place for this type of post. As I indicated to you in a private post, I personally do not believe that we here on the histonet can be considered "clients" simply because you have posted here or even if one of us has asked about a position. I feel that the only people that you can consider "clients" are those who have signed up for your services either as an employer or employee that you have placed. I find your post offensive and unprofessional. You have every right to contact your true clients and elicit their support in your endeavors but I feel that posting here is inappropriate. Susan Eric Dye wrote: Hi Histonetters, I just wanted to give you a warning about Pam Barker/Relia Inc, and ask for your help. Here at Ategra, we used to have an employee named Pam Barker. She resigned from Ategra last year (Jan 20 2005) and started her own company named Relia LLC. Since she resigned we here at Ategra were alarmed to find that Pam Barker has contacted Ategra clients and candidates in violation of her non-compete agreement. The court has agreed and has issued an Injunction (like a restraining order) against Pam Barker/Relia LLC. This Injunction (like a restraining order) specifically prohibits Pam Barker from contacting Ategra clients/candidates (including prospective clients/candidates) From Jan 05 2006 until Aug 8th 2006 (if not later) Because I strive to keep Ategra's contact lists strictly confidential, I would like to ask a favor of you: The favor I ask is this : if you have been contacted by Pam Barker OR Relia LLC if you could PLEASE LET ME KNOW ? Your name will be kept confidential and no other involvement from you or your firm is necessary. All I need to know is the appx date she has contacted you. Thank You in Advance For Your Help Eric Dye, PS: If you have any questions about this, call me at 800-466-9919 ext 223 PSS: If you need to see a copy of the Injunction (restraining order) against Pam Barker, click here: http://www.ategra.com/injunction.html PSSS: I have openings for HistoTechs throughout the US. If you are thinking of a making career move, feel free to call me. ---------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd Winter Park, FL 32792-6721 VOICE: 800-466-9919 ext 223 FAX: 407-671-6075 EMAIL: Eric@ategra.com ---------------------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From pmcardle <@t> ebsciences.com Thu Feb 9 08:52:34 2006 From: pmcardle <@t> ebsciences.com (Phil McArdle) Date: Thu Feb 9 08:52:44 2006 Subject: [Histonet] Microwave processors In-Reply-To: <000301c62d84$f998ee00$4f13e980@PDS11> References: <000301c62d84$f998ee00$4f13e980@PDS11> Message-ID: <43EB5732.4010809@ebsciences.com> Hi Brian: Please forgive a "vendor" response; there are standalone microwave tissue processors that work in conjunction with your existing work flow (and which, with vacuum, can accommodate relatively thick fatty specimens), and autoprocessors that also incorporate microwave technology. For more information, including protocols, etc., you can visit the "library" section of www.ebsciences.com. Hope this helps! Best regards, Phil McArdle -- Phil McArdle Microwave Product Manager Energy Beam Sciences, Inc. Tel: 800.992.9037 x 341 Fax: 860.653.0422 PMcardle@ebsciences.com www.ebsciences.com "ADDING BRILLIANCE TO YOUR VISION" Brian Chelack wrote: > Can any one provide me with information regarding microwave tissue > processors? What is the quality of the sections derived from microwave > processors compared to regular processors? Are there any reliability issues? > Do you need to trim in very thin tissues? Anything else? > > > Thanks > > Brian Chelack > Prairie Diagnostic Services > 2604-52 Campus Drive > Saskatoon SK > S7N 5B4 > 306-966-7241 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From DKAEK <@t> coloplast.com Thu Feb 9 08:53:27 2006 From: DKAEK <@t> coloplast.com (Annette Ekblond) Date: Thu Feb 9 08:53:40 2006 Subject: [Histonet] epithelial differentiation-paraffin embedded tissue Message-ID: Dear Histonet subscribers We find it difficult to identify the following antigens in paraffin embedded epithelium: Laminin 1 Laminin 5 Transglutaminase 1 Collagen VII It works for us in cryopreserved specimens - but not in paraffin....... Does anyone have any suggestions - or perhaps even successfull protocols to solve this problem? Sincerely Annette Coloplast Research Denmark From Barry.R.Rittman <@t> uth.tmc.edu Thu Feb 9 09:07:20 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Thu Feb 9 09:07:25 2006 Subject: [Histonet] Asking for your help ... Message-ID: Eric I agree that the Histonet is no place for this type of posting. What were you thinking? If you have an issue with this individual then there are other recourses including legal avenues. You are asking us to become involved in this and it is not appropriate. My first inclination in reading your note was to support the individual that you refer to - in a court of law both sides would have the opportunity to present their case and have it considered. I think that you have done a disservice to Ategra Systems by this posting. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Horn, Hazel V Sent: Thursday, February 09, 2006 8:48 AM To: Histology SLU; Eric Dye; Histonetters Subject: RE: [Histonet] Asking for your help ... You are absolutely correct Susan. This post offended me and this list cannot be considered Eric's client. Hazel -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histology SLU Sent: Thursday, February 09, 2006 7:20 AM To: Eric Dye; Histonetters Subject: Re: [Histonet] Asking for your help ... Eric: I do not believe that this is the place for this type of post. As I indicated to you in a private post, I personally do not believe that we here on the histonet can be considered "clients" simply because you have posted here or even if one of us has asked about a position. I feel that the only people that you can consider "clients" are those who have signed up for your services either as an employer or employee that you have placed. I find your post offensive and unprofessional. You have every right to contact your true clients and elicit their support in your endeavors but I feel that posting here is inappropriate. Susan Eric Dye wrote: Hi Histonetters, I just wanted to give you a warning about Pam Barker/Relia Inc, and ask for your help. Here at Ategra, we used to have an employee named Pam Barker. She resigned from Ategra last year (Jan 20 2005) and started her own company named Relia LLC. Since she resigned we here at Ategra were alarmed to find that Pam Barker has contacted Ategra clients and candidates in violation of her non-compete agreement. The court has agreed and has issued an Injunction (like a restraining order) against Pam Barker/Relia LLC. This Injunction (like a restraining order) specifically prohibits Pam Barker from contacting Ategra clients/candidates (including prospective clients/candidates) From Jan 05 2006 until Aug 8th 2006 (if not later) Because I strive to keep Ategra's contact lists strictly confidential, I would like to ask a favor of you: The favor I ask is this : if you have been contacted by Pam Barker OR Relia LLC if you could PLEASE LET ME KNOW ? Your name will be kept confidential and no other involvement from you or your firm is necessary. All I need to know is the appx date she has contacted you. Thank You in Advance For Your Help Eric Dye, PS: If you have any questions about this, call me at 800-466-9919 ext 223 PSS: If you need to see a copy of the Injunction (restraining order) against Pam Barker, click here: http://www.ategra.com/injunction.html PSSS: I have openings for HistoTechs throughout the US. If you are thinking of a making career move, feel free to call me. ---------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd Winter Park, FL 32792-6721 VOICE: 800-466-9919 ext 223 FAX: 407-671-6075 EMAIL: Eric@ategra.com ---------------------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------ ------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ======================================================================== ====== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nancy.troiano <@t> yale.edu Thu Feb 9 09:42:16 2006 From: nancy.troiano <@t> yale.edu (Nancy W. Troiano) Date: Thu Feb 9 09:42:07 2006 Subject: [Histonet] plastic embedded bone sections Message-ID: <5.2.1.1.2.20060209103821.00c3ace8@email.med.yale.edu> We routinely cut MMA embedded mouse and rat tibias using a 160 mm D-profile tungsten carbide microtome blade. A good source of excellent blades/resharpening service is Dorn/Hart microedge (Ken Hart), tel #630-832-3843. Ken is also a great source of information as to the proper blade angles, blades, etc for your application. I am not familiar with the microm microtomes but we use a Leica 2165 motorized microtome (you definitely need a motorized unit) and set our blade angle at approximately 2.5 to 3 (number setiing on side of knife holder). You may have to change your angles as the blade is resharpened. From lchung <@t> ppmh.org Thu Feb 9 09:43:02 2006 From: lchung <@t> ppmh.org (Chung, Luong) Date: Thu Feb 9 09:43:28 2006 Subject: [Histonet] Asking for your help ... Message-ID: Thank you Susan. Ditto. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Pam Marcum Sent: Thursday, February 09, 2006 9:42 AM To: Histology SLU; Eric Dye; Histonetters Subject: Re: [Histonet] Asking for your help ... Thank you Susan. Well said and I agree. This is a private business matter between two companies and thier signed clients not all of HistoNet. While we appreciate knowing where the jobs we are not clients because we see and read the ads placed here. Pam Marcum -------------- Original message ---------------------- From: Histology SLU > Eric: > > I do not believe that this is the place for this type of post. As I indicated > to you in a private post, I personally do not believe that we here on the > histonet can be considered "clients" simply because you have posted here or even > if one of us has asked about a position. I feel that the only people that you > can consider "clients" are those who have signed up for your services either as > an employer or employee that you have placed. I find your post offensive and > unprofessional. You have every right to contact your true clients and elicit > their support in your endeavors but I feel that posting here is inappropriate. > > Susan > > Eric Dye wrote: > Hi Histonetters, I just wanted to give you a warning about Pam Barker/Relia > Inc, and ask for your help. > > Here at Ategra, we used to have an employee named Pam Barker. She resigned from > Ategra last year (Jan 20 2005) and started her own company named Relia LLC. > > Since she resigned we here at Ategra were alarmed to find that Pam Barker has > contacted Ategra clients and candidates in violation of her non-compete > agreement. > > The court has agreed and has issued an Injunction (like a restraining order) > against Pam Barker/Relia LLC. This Injunction (like a restraining order) > specifically prohibits Pam Barker from contacting Ategra clients/candidates > (including prospective clients/candidates) From Jan 05 2006 until Aug 8th 2006 > (if not later) > > Because I strive to keep Ategra's contact lists strictly confidential, I would > like to ask a favor of you: > > The favor I ask is this : if you have been contacted by Pam Barker OR Relia LLC > if you could PLEASE LET ME KNOW ? > > Your name will be kept confidential and no other involvement from you or your > firm is necessary. All I need to know is the appx date she has contacted you. > > Thank You in Advance For Your Help > > Eric Dye, > > PS: If you have any questions about this, call me at 800-466-9919 ext 223 > > PSS: If you need to see a copy of the Injunction (restraining order) against Pam > Barker, click here: http://www.ategra.com/injunction.html > > PSSS: I have openings for HistoTechs throughout the US. If you are thinking of a > making career move, feel free to call me. > > ---------------------------------------------------- > Ategra Systems Inc > Specialists in Permanent & Contract Staffing > 7085 University Blvd > Winter Park, FL 32792-6721 > > VOICE: 800-466-9919 ext 223 > FAX: 407-671-6075 > EMAIL: Eric@ategra.com > > ---------------------------------------------------- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > --------------------------------- > Relax. Yahoo! Mail virus scanning helps detect nasty viruses! > _______________________________________________ > 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 ----------------------------------------- Disclaimer: The HIPAA Final Privacy Rule requires covered entities to safeguard certain Protected Health Information (PHI) related to a person's healthcare. Information being faxed to you may include PHI after appropriate authorization from the patient or under circumstances that do not require patient authorization. You, the recipient, are obligated to maintain PHI in a safe and secure manner. You may not re-disclose without additional patient consent or as required by law. Unauthorized re-disclosure or failure to safeguard PHI could subject you to penalties described in federal (HIPAA) and state law. If you the reader of this message are not the intended recipient, or the employee or agent responsible to deliver it to the intended recipient, please notify us immediately and destroy the related message. From tkngflght <@t> yahoo.com Thu Feb 9 09:59:15 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Thu Feb 9 09:59:24 2006 Subject: [Histonet] the job of a good recruiter-- In-Reply-To: Message-ID: <20060209155916.24397.qmail@web50901.mail.yahoo.com> Hi all- As a 23 year tech, with ten years as a travel tech, I have worked with nearly all of the recruiters and headhunters, including those concerned in the recent discussions. This concern with restricting access and bottom-line/assets is WRONG. WE ARE NOT CATTLE. We have the right to choose who we will and won't work with. I've been a manager working 12 hour days because I couldn't find the right tech for my hospital. I've been the tech looking for the next step in my career without the resources to find that needle in a haystack and know that I got the RIGHT job for my career and my family. Recruiters and headhunters do have a place in Lab Science. These are the EXACT reasons I started an agency--to help each tech who comes to me --to help them meet their professional and FAMILY goals--not to push them into the next available job or restrict their ability to work with a company that might be able to help them better than mine. I even REFER my candidates to other agencies if I can't help them (including Pam when she was with Ategra because she is a good recruiter)! I've had recruiters call repeatedly and try to push me into Med Tech jobs after I'd spent my time explaining that an HT/HTL is a TISSUE person: we don't do blood. I've had calls asking me to do phlebotomy for $7 an hour....these agents weren't thinking of my needs at all. I've had calls from hospitals where I'd been submitted without my knowledge--they wasted that HR person's time. Histonet is a place for sharing information to better our science--and to help each other be better techs. If the recruiters and the headhunters get their heads set right --that their job is to help PEOPLE--both techs and people who need them, the rest will fall into place all by itself without the need for legal intervention. Okay, I'll get off my soapbox now...thanks for listening :-) Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one tech at a time. admin@fullstaff.org 281.852.9457 281.883.7704 From anh2006 <@t> med.cornell.edu Thu Feb 9 10:16:08 2006 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Thu Feb 9 10:16:21 2006 Subject: [Histonet] In situ RT-PCR - is it being done? Message-ID: Dear All, Is anyone routinely performing the IS-RTPCR technique on tissue sections? What is the histology field's current thought on the utility and technical application of the procedure? More practically, is it do-able and reproducible and BELIEVABLE? Any anecdotal information is appreciated. I have several books/references on the subject already, I am more interested in stories from people in the lab. Thanks, Andrea -- From dusko.trajkovic <@t> pfizer.com Thu Feb 9 10:34:43 2006 From: dusko.trajkovic <@t> pfizer.com (Trajkovic, Dusko) Date: Thu Feb 9 10:34:58 2006 Subject: [Histonet] Asking for your help ... Message-ID: <3AD0BD3142459B4E9B12CBEAFF2B89B2D8975E@lajamrexm01.amer.pfizer.com> I wholeheartedly agree with Barry! Dusko -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rittman, Barry R Sent: Thursday, February 09, 2006 7:07 AM To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Asking for your help ... Eric I agree that the Histonet is no place for this type of posting. What were you thinking? If you have an issue with this individual then there are other recourses including legal avenues. You are asking us to become involved in this and it is not appropriate. My first inclination in reading your note was to support the individual that you refer to - in a court of law both sides would have the opportunity to present their case and have it considered. I think that you have done a disservice to Ategra Systems by this posting. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Horn, Hazel V Sent: Thursday, February 09, 2006 8:48 AM To: Histology SLU; Eric Dye; Histonetters Subject: RE: [Histonet] Asking for your help ... You are absolutely correct Susan. This post offended me and this list cannot be considered Eric's client. Hazel -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histology SLU Sent: Thursday, February 09, 2006 7:20 AM To: Eric Dye; Histonetters Subject: Re: [Histonet] Asking for your help ... Eric: I do not believe that this is the place for this type of post. As I indicated to you in a private post, I personally do not believe that we here on the histonet can be considered "clients" simply because you have posted here or even if one of us has asked about a position. I feel that the only people that you can consider "clients" are those who have signed up for your services either as an employer or employee that you have placed. I find your post offensive and unprofessional. You have every right to contact your true clients and elicit their support in your endeavors but I feel that posting here is inappropriate. Susan Eric Dye wrote: Hi Histonetters, I just wanted to give you a warning about Pam Barker/Relia Inc, and ask for your help. Here at Ategra, we used to have an employee named Pam Barker. She resigned from Ategra last year (Jan 20 2005) and started her own company named Relia LLC. Since she resigned we here at Ategra were alarmed to find that Pam Barker has contacted Ategra clients and candidates in violation of her non-compete agreement. The court has agreed and has issued an Injunction (like a restraining order) against Pam Barker/Relia LLC. This Injunction (like a restraining order) specifically prohibits Pam Barker from contacting Ategra clients/candidates (including prospective clients/candidates) From Jan 05 2006 until Aug 8th 2006 (if not later) Because I strive to keep Ategra's contact lists strictly confidential, I would like to ask a favor of you: The favor I ask is this : if you have been contacted by Pam Barker OR Relia LLC if you could PLEASE LET ME KNOW ? Your name will be kept confidential and no other involvement from you or your firm is necessary. All I need to know is the appx date she has contacted you. Thank You in Advance For Your Help Eric Dye, PS: If you have any questions about this, call me at 800-466-9919 ext 223 PSS: If you need to see a copy of the Injunction (restraining order) against Pam Barker, click here: http://www.ategra.com/injunction.html PSSS: I have openings for HistoTechs throughout the US. If you are thinking of a making career move, feel free to call me. ---------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd Winter Park, FL 32792-6721 VOICE: 800-466-9919 ext 223 FAX: 407-671-6075 EMAIL: Eric@ategra.com ---------------------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------ ------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ======================================================================== ====== _______________________________________________ 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 ---------------------------------------------------------------------- LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From Eric <@t> ategra.com Thu Feb 9 11:02:36 2006 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Thu Feb 9 10:51:50 2006 Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for your help ...) Message-ID: Please accept my sincerest apologies for posting the email titled "Asking For Your Help" to the HistoNet. Those who commented are indeed correct: HistoNet is an inappropriate place for that type of posting. My list of HistoTechs is quite large. When a coworker and I built the list of prospective candidates, it was our intent to specifically exclude the HistoNet. Unfortunately, we goofed and the HistoNet got included. Once again accept my apology. Thank You for your understanding. Eric Dye, Sr HistoTech Recruiter PS: If you have any questions about this, call me at 800-466-9919 ext 223 ------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd Winter Park, FL 32792-6721 VOICE: 800-466-9919 ext 223 FAX: 407-671-6075 EMAIL: Eric@ategra.com ------------------------------------------------------- From Melissa.Gonzalez <@t> cellgenesys.com Thu Feb 9 10:28:43 2006 From: Melissa.Gonzalez <@t> cellgenesys.com (Melissa Gonzalez) Date: Thu Feb 9 10:53:10 2006 Subject: [Histonet] RE: Slide scanner for Fluorescence Message-ID: Kim, Look up CompuCyte. I recently went to one of their talks, and their technology is amazing. Pricey, but you get a lot out of the system. It is based on laser scanning instead of traditional arc lamps and it has the capability to load slides for overnight/weekend runs. In addition to images, you can also acquire graphs and population statistics about your stained slides. It is the first system for fluorescent based applications that I've seen that is so sensitive. Cheers, Melissa Melissa A. Gonzalez R&D Histology Cell Genesys, South San Francisco, CA 94080 Message: 20 Date: Thu, 9 Feb 2006 06:31:03 -0800 From: "Trajkovic, Dusko" Subject: RE: [Histonet] Slide scanner for Quantum Dot Fluorescence To: "Kim Merriam" , "Histonet" Message-ID: <3AD0BD3142459B4E9B12CBEAFF2B89B2D89759@lajamrexm01.amer.pfizer.com> Content-Type: text/plain; charset="us-ascii" Try contacting Ventana Medical Systems. 1-800-227-2155 Dusko Trajkovic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Merriam Sent: Thursday, February 09, 2006 4:53 AM To: Histonet Subject: [Histonet] Slide scanner for Quantum Dot Fluorescence Hello All, I am looking for a slide scanner (captures images of entire tissue sections on slides) that has fluorescent capabilities. We currently have an Aperio slide scanner for our routine light microscopy slides, we are very happy with it, but we will be branching out into Quantum Dots and would like a scanner that can do the same thing for fluorescent-stained slides. Does anyone have any ideas or recommendations? Thanks, Kim Kim Merriam Novartis Cambridge, MA From sluhisto <@t> yahoo.com Thu Feb 9 10:57:49 2006 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Thu Feb 9 10:57:57 2006 Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for your help ...) In-Reply-To: Message-ID: <20060209165749.80460.qmail@web51010.mail.yahoo.com> Eric: Thank you for your post. Your retraction is appreciated. Susan Eric Dye wrote: Please accept my sincerest apologies for posting the email titled "Asking For Your Help" to the HistoNet. Those who commented are indeed correct: HistoNet is an inappropriate place for that type of posting. My list of HistoTechs is quite large. When a coworker and I built the list of prospective candidates, it was our intent to specifically exclude the HistoNet. Unfortunately, we goofed and the HistoNet got included. Once again accept my apology. Thank You for your understanding. Eric Dye, Sr HistoTech Recruiter PS: If you have any questions about this, call me at 800-466-9919 ext 223 ------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd Winter Park, FL 32792-6721 VOICE: 800-466-9919 ext 223 FAX: 407-671-6075 EMAIL: Eric@ategra.com ------------------------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! From DDDeltour <@t> mar.med.navy.mil Thu Feb 9 11:05:14 2006 From: DDDeltour <@t> mar.med.navy.mil (DDDeltour@mar.med.navy.mil) Date: Thu Feb 9 11:05:43 2006 Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for you r help ...) Message-ID: <3F500F8B416C554EBB21FF16642F72E959CDBF@marxchg03.mar.med.navy.mil> Eric, If this were true then your original message would not have said..." Hi Histonetters". Admit that you used bad judgment and not accidentally sent the message to Histonet. Thanks -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Eric Dye Sent: Thursday, February 09, 2006 12:03 PM To: Histonetters Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for your help ...) Please accept my sincerest apologies for posting the email titled "Asking For Your Help" to the HistoNet. Those who commented are indeed correct: HistoNet is an inappropriate place for that type of posting. My list of HistoTechs is quite large. When a coworker and I built the list of prospective candidates, it was our intent to specifically exclude the HistoNet. Unfortunately, we goofed and the HistoNet got included. Once again accept my apology. Thank You for your understanding. Eric Dye, Sr HistoTech Recruiter PS: If you have any questions about this, call me at 800-466-9919 ext 223 ------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd Winter Park, FL 32792-6721 VOICE: 800-466-9919 ext 223 FAX: 407-671-6075 EMAIL: Eric@ategra.com ------------------------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From anh2006 <@t> med.cornell.edu Thu Feb 9 11:05:34 2006 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Thu Feb 9 11:05:46 2006 Subject: [Histonet] B-Gal endogenous expression Message-ID: Does anyone have references or anecdotal experience as to where mammalian B-Gal is endogenously expressed if anywhere in mouse or human? Thanks, Andrea -- From DDDeltour <@t> mar.med.navy.mil Thu Feb 9 11:06:41 2006 From: DDDeltour <@t> mar.med.navy.mil (DDDeltour@mar.med.navy.mil) Date: Thu Feb 9 11:07:04 2006 Subject: [Histonet] the job of a good recruiter-- Message-ID: <3F500F8B416C554EBB21FF16642F72E959CDC0@marxchg03.mar.med.navy.mil> Cheryl, Since you want to put it out there about recruiters, I hope you don't mind if I ask you a few question regarding recruiters since I will be changing jobs soon. 1) What can a recruiter do for me that I can't do for myself? 2) Why do recruiters want me to give them my SSN? 3) Why do they ask me if I applied elsewhere and if I am in contact with other recruiters? 4) If I apply for a job through a hospital website vs. a recruiter will my starting pay be different due to recruiter cost? I have been contacted by SEVERAL recruiters since I posted my resume on a few of the job sites. I have yet to be impressed. Please tell me why I need to deal with a recruiter??? I am not being "smart" I just am curious. Thanks Douglas D. Deltour HT(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Cheryl Sent: Thursday, February 09, 2006 10:59 AM To: histonet@lists.utsouthwestern.edu Subject: [POSSIBLE SPAM!!] [Histonet] the job of a good recruiter-- Importance: Low Hi all- As a 23 year tech, with ten years as a travel tech, I have worked with nearly all of the recruiters and headhunters, including those concerned in the recent discussions. This concern with restricting access and bottom-line/assets is WRONG. WE ARE NOT CATTLE. We have the right to choose who we will and won't work with. I've been a manager working 12 hour days because I couldn't find the right tech for my hospital. I've been the tech looking for the next step in my career without the resources to find that needle in a haystack and know that I got the RIGHT job for my career and my family. Recruiters and headhunters do have a place in Lab Science. These are the EXACT reasons I started an agency--to help each tech who comes to me --to help them meet their professional and FAMILY goals--not to push them into the next available job or restrict their ability to work with a company that might be able to help them better than mine. I even REFER my candidates to other agencies if I can't help them (including Pam when she was with Ategra because she is a good recruiter)! I've had recruiters call repeatedly and try to push me into Med Tech jobs after I'd spent my time explaining that an HT/HTL is a TISSUE person: we don't do blood. I've had calls asking me to do phlebotomy for $7 an hour....these agents weren't thinking of my needs at all. I've had calls from hospitals where I'd been submitted without my knowledge--they wasted that HR person's time. Histonet is a place for sharing information to better our science--and to help each other be better techs. If the recruiters and the headhunters get their heads set right --that their job is to help PEOPLE--both techs and people who need them, the rest will fall into place all by itself without the need for legal intervention. Okay, I'll get off my soapbox now...thanks for listening :-) Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one tech at a time. admin@fullstaff.org 281.852.9457 281.883.7704 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dusko.trajkovic <@t> pfizer.com Thu Feb 9 11:08:35 2006 From: dusko.trajkovic <@t> pfizer.com (Trajkovic, Dusko) Date: Thu Feb 9 11:08:47 2006 Subject: [Histonet] Bielschowsky and Gallyas Specificity Message-ID: <3AD0BD3142459B4E9B12CBEAFF2B89B2D8975F@lajamrexm01.amer.pfizer.com> Good Morning Everyone, I'm posting this for a colleague of mine. Thank you Dusko Trajkovic Hi Dusko, Good morning! As I mentioned before, I am interested to know more details about the specificity of these two (Bielschowsky & Gallyas) staining methods. i.e., if more is known about the exact "protein" or "cellular structure" they stain. Can you ask this on the histonet for me? Thanks a lot! Jessie ---------------------------------------------------------------------- LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From Linke_Noelle <@t> Allergan.com Thu Feb 9 11:13:10 2006 From: Linke_Noelle <@t> Allergan.com (Linke_Noelle) Date: Thu Feb 9 11:13:40 2006 Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for your help ...) Message-ID: You people need to chill out and get over this....... If you don't like what you read, the 'delete' key is right there for you to use..... Who are you to judge??? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DDDeltour@mar.med.navy.mil Sent: Thursday, February 09, 2006 9:05 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Public APOLOGY to the HistoNet (re: Asking for your help ...) Eric, If this were true then your original message would not have said..." Hi Histonetters". Admit that you used bad judgment and not accidentally sent the message to Histonet. Thanks -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Eric Dye Sent: Thursday, February 09, 2006 12:03 PM To: Histonetters Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for your help ...) Please accept my sincerest apologies for posting the email titled "Asking For Your Help" to the HistoNet. Those who commented are indeed correct: HistoNet is an inappropriate place for that type of posting. My list of HistoTechs is quite large. When a coworker and I built the list of prospective candidates, it was our intent to specifically exclude the HistoNet. Unfortunately, we goofed and the HistoNet got included. Once again accept my apology. Thank You for your understanding. Eric Dye, Sr HistoTech Recruiter PS: If you have any questions about this, call me at 800-466-9919 ext 223 ------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd Winter Park, FL 32792-6721 VOICE: 800-466-9919 ext 223 FAX: 407-671-6075 EMAIL: Eric@ategra.com ------------------------------------------------------- _______________________________________________ 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 From RRA <@t> Stowers-Institute.org Thu Feb 9 11:14:10 2006 From: RRA <@t> Stowers-Institute.org (Allen, Rhonda) Date: Thu Feb 9 11:14:31 2006 Subject: [Histonet] RE: Ihc bone staining Message-ID: > > Does anyone know if it could possibly take longer(or more concentrated > antibody) to stain the bone in a tissue section(mouse head, newborn > pup, not decalcified), vs. the other tissue components(brain, skin). > In order to get nice staining in the bone, seems like we are starting > to see background coming up in the brain. Wondering if this is normal > for some antibodies. > Thanks in advance, > Rhonda Allen BA HT(ASCP)HTL, QIHC > Histotechnology Specialist II > Stowers Institute > 1000 E. 50th Street > Kansas City, MO 64110 > 816-926-4305 > > From pjfnefro <@t> duke.edu Thu Feb 9 11:19:54 2006 From: pjfnefro <@t> duke.edu (Pat Flannery) Date: Thu Feb 9 11:20:02 2006 Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for your help ...) In-Reply-To: References: Message-ID: Thank you, now let's gang up on DDDeltour (he's a sailor, he can take it). :-) -- -Patrick J. (Pat) Flannery Division of Nephrology (that's kidneys to you) Box 3014 (that's NOT "PO" just "Box") Duke University Medical Center Durham, NC 27710 E-mail: pjfnefro@duke.edu (preferred) FLANN002@MC.DUKE.EDU (also works) Voice: (919)660-6863 Fax: (919)684-2929 On Feb 9, 2006, at 12:13 PM, Linke_Noelle wrote: >> You people need to chill out and get over this....... If you >> don't like >> what you read, the 'delete' key is right there for you to >> use..... Who >> are you to judge??? >> From pjfnefro <@t> duke.edu Thu Feb 9 11:25:30 2006 From: pjfnefro <@t> duke.edu (Pat Flannery) Date: Thu Feb 9 11:25:39 2006 Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for you r help ...) In-Reply-To: <3F500F8B416C554EBB21FF16642F72E959CDBF@marxchg03.mar.med.navy.mil> References: <3F500F8B416C554EBB21FF16642F72E959CDBF@marxchg03.mar.med.navy.mil> Message-ID: <657FAD46-DB52-4182-A2AD-0A61384195FB@duke.edu> Gee, let's not only reject his apology, let's publicly flog the guy. Boy, you're pretty harsh on this list! -Pat Flannery pjfnefro@duke.edu On Feb 9, 2006, at 12:05 PM, DDDeltour@mar.med.navy.mil wrote: > Eric, > If this were true then your original message would not have > said..." Hi > Histonetters". Admit that you used bad judgment and not > accidentally sent > the message to Histonet. Thanks > > From Steve_Dalessandro <@t> rsh.net Thu Feb 9 11:30:14 2006 From: Steve_Dalessandro <@t> rsh.net (Steve_Dalessandro@rsh.net) Date: Thu Feb 9 11:30:24 2006 Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for you r help ...) Message-ID: No...not harsh.. they just don't have that much work to do. From DDDeltour <@t> mar.med.navy.mil Thu Feb 9 11:30:22 2006 From: DDDeltour <@t> mar.med.navy.mil (DDDeltour@mar.med.navy.mil) Date: Thu Feb 9 11:30:41 2006 Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for you r help ...) Message-ID: <3F500F8B416C554EBB21FF16642F72E959CDC1@marxchg03.mar.med.navy.mil> Bring it! :) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat Flannery Sent: Thursday, February 09, 2006 12:20 PM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Public APOLOGY to the HistoNet (re: Asking for your help ...) Thank you, now let's gang up on DDDeltour (he's a sailor, he can take it). :-) -- -Patrick J. (Pat) Flannery Division of Nephrology (that's kidneys to you) Box 3014 (that's NOT "PO" just "Box") Duke University Medical Center Durham, NC 27710 E-mail: pjfnefro@duke.edu (preferred) FLANN002@MC.DUKE.EDU (also works) Voice: (919)660-6863 Fax: (919)684-2929 On Feb 9, 2006, at 12:13 PM, Linke_Noelle wrote: >> You people need to chill out and get over this....... If you >> don't like >> what you read, the 'delete' key is right there for you to >> use..... Who >> are you to judge??? >> _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DDDeltour <@t> mar.med.navy.mil Thu Feb 9 11:37:39 2006 From: DDDeltour <@t> mar.med.navy.mil (DDDeltour@mar.med.navy.mil) Date: Thu Feb 9 11:37:59 2006 Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for you r help ...) Message-ID: <3F500F8B416C554EBB21FF16642F72E959CDC2@marxchg03.mar.med.navy.mil> Pat, It is call integrity. Forgive me if I am out of line and it doesn't exist anymore. :) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat Flannery Sent: Thursday, February 09, 2006 12:26 PM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Public APOLOGY to the HistoNet (re: Asking for you r help ...) Gee, let's not only reject his apology, let's publicly flog the guy. Boy, you're pretty harsh on this list! -Pat Flannery pjfnefro@duke.edu On Feb 9, 2006, at 12:05 PM, DDDeltour@mar.med.navy.mil wrote: > Eric, > If this were true then your original message would not have > said..." Hi > Histonetters". Admit that you used bad judgment and not > accidentally sent > the message to Histonet. Thanks > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Thu Feb 9 11:46:08 2006 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Thu Feb 9 11:44:33 2006 Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for you rhelp ...) Message-ID: I agree to the harsh bit. Two have been "offended" however, so perhaps we should organise a public march, with banners demanding that the perpetrator be beheaded. There is a historical precedence. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Pat Flannery [mailto:pjfnefro@duke.edu] Sent: 09 February 2006 17:26 To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Public APOLOGY to the HistoNet (re: Asking for you rhelp ...) Gee, let's not only reject his apology, let's publicly flog the guy. Boy, you're pretty harsh on this list! -Pat Flannery pjfnefro@duke.edu On Feb 9, 2006, at 12:05 PM, DDDeltour@mar.med.navy.mil wrote: > Eric, > If this were true then your original message would not have > said..." Hi > Histonetters". Admit that you used bad judgment and not > accidentally sent > the message to Histonet. Thanks > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RebeNoel <@t> aol.com Thu Feb 9 11:46:45 2006 From: RebeNoel <@t> aol.com (RebeNoel@aol.com) Date: Thu Feb 9 11:47:58 2006 Subject: [Histonet] the job of a good recruiter-- Message-ID: <12b.6dccccf3.311cda05@aol.com> I have been a recruiter for over 10 years and this has been my experience?. ) What can a recruiter do for me that I can't do for myself? A recruiter can do a couple of things- Recruiter spends years building relationships with clients built on trust and creditability. The client that a particular recruiter has a strong relationship with feels that when he/she calls them about a potential candidate, that the candidate is not only has the skills and experience but will also be a good fit in terms of soft skills/personality. I have clients that I know very well and I know what type of person will be successful in their environment. Second- Often when a client posts an open position on their hospital website or a job board the client literally will get hundreds of resumes. Really qualified people can often get overlooked only because their resume is in a stack of 200 other resumes. A successful recruiter will often have a contact person at the facility who they can call and discuss your background in detail. This gives the candidate a great advantage. 2) Why do recruiters want me to give them my SSN? SSN should only be given if you are going to do contract work or a background check is required for a certain position you are applying for? I myself do not require anyone to give me their SSN number unless it is absolutely necessary. 3) Why do they ask me if I applied elsewhere and if I am in contact with other recruiters? Recruiters ask you where you have applied because if a client receives your resume through a hospital web site or job board the recruiter no long has the right to collect a fee for the position you applied for, even if the client did not initially call you.. it make no difference. If they had your resume in a filing cabinet you still are considered a candidate that has already been submitted. Another reason is that it gives a recruiter an idea of what types of positions are appealing to you. And finally some shady recruiters use this technique to find out who is hiring so they can call and try and get the job order. 4) If I apply for a job through a hospital web site vs. a recruiter will my starting pay be different due to recruiter cost? Starting pay could be effected because if a recruiter has been working with a particular client for a while the recruiter knows what that hospital typically pays. An example would be if ABC hospital typically pays Histotech 40K and you put on your application that 35K is what you are looking to make there is a good chance 35K is what you would be paid. A recruiter knowing the history of salaries for that hospital would negotiate of 40K salary. Rebecca Noel Executive Match Personnel 919-601-1946 From tkngflght <@t> yahoo.com Thu Feb 9 11:52:18 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Thu Feb 9 11:52:30 2006 Subject: [Histonet] email generation and working sailors... In-Reply-To: Message-ID: <20060209175218.68392.qmail@web50910.mail.yahoo.com> Hey everyone-- Eric's database most probably did as he said--some of his emails run through a program that auto-generates groups of outgoing messages--including the greeting on the top line. I received this message both through Histonet and my personal email as I am on his database seeking an admin job. I appreciate that he apologized. And Mr. Deltour appears to have been put through the wringer by several recruiters: he's been on the short end of the stick it seems. Let's cut the fella some slack--we've all been there!! Okay everybody--back to work!! (grin!!) Cheryl Pat Flannery wrote: Thank you, now let's gang up on DDDeltour (he's a sailor, he can take it). :-) -- -Patrick J. (Pat) Flannery Division of Nephrology (that's kidneys to you) Box 3014 (that's NOT "PO" just "Box") Duke University Medical Center Durham, NC 27710 E-mail: pjfnefro@duke.edu (preferred) FLANN002@MC.DUKE.EDU (also works) Voice: (919)660-6863 Fax: (919)684-2929 On Feb 9, 2006, at 12:13 PM, Linke_Noelle wrote: >> You people need to chill out and get over this....... If you >> don't like >> what you read, the 'delete' key is right there for you to >> use..... Who >> are you to judge??? >> _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tkngflght <@t> yahoo.com Thu Feb 9 11:58:14 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Thu Feb 9 11:58:26 2006 Subject: [Histonet] (no subject) In-Reply-To: <12b.6dccccf3.311cda05@aol.com> Message-ID: <20060209175815.31072.qmail@web50905.mail.yahoo.com> RebeNoel@aol.com wrote: I have been a recruiter for over 10 years and this has been my experience???. ) What can a recruiter do for me that I can't do for myself? A recruiter can do a couple of things- Recruiter spends years building relationships with clients built on trust and creditability. The client that a particular recruiter has a strong relationship with feels that when he/she calls them about a potential candidate, that the candidate is not only has the skills and experience but will also be a good fit in terms of soft skills/personality. I have clients that I know very well and I know what type of person will be successful in their environment. Second- Often when a client posts an open position on their hospital website or a job board the client literally will get hundreds of resumes. Really qualified people can often get overlooked only because their resume is in a stack of 200 other resumes. A successful recruiter will often have a contact person at the facility who they can call and discuss your background in detail. This gives the candidate a great advantage. 2) Why do recruiters want me to give them my SSN? SSN should only be given if you are going to do contract work or a background check is required for a certain position you are applying for??? I myself do not require anyone to give me their SSN number unless it is absolutely necessary. 3) Why do they ask me if I applied elsewhere and if I am in contact with other recruiters? Recruiters ask you where you have applied because if a client receives your resume through a hospital web site or job board the recruiter no long has the right to collect a fee for the position you applied for, even if the client did not initially call you.. it make no difference. If they had your resume in a filing cabinet you still are considered a candidate that has already been submitted. Another reason is that it gives a recruiter an idea of what types of positions are appealing to you. And finally some shady recruiters use this technique to find out who is hiring so they can call and try and get the job order. 4) If I apply for a job through a hospital web site vs. a recruiter will my starting pay be different due to recruiter cost? Starting pay could be effected because if a recruiter has been working with a particular client for a while the recruiter knows what that hospital typically pays. An example would be if ABC hospital typically pays Histotech 40K and you put on your application that 35K is what you are looking to make there is a good chance 35K is what you would be paid. A recruiter knowing the history of salaries for that hospital would negotiate of 40K salary. Rebecca Noel Executive Match Personnel 919-601-1946 From Sue.Kapoor <@t> uhsi.org Thu Feb 9 12:17:25 2006 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Thu Feb 9 12:16:48 2006 Subject: [Histonet] water for frozen section staining Message-ID: Hello histonetters, We have a debate going in my lab as to using warm/hot or cold running water when staining frozen sections. One side states warm/hot water "blues" the hematoxylin...the other sides feels the cold water keeps the sections from falling off. What does everyone in histo land do and think??? thanks for your replies :) Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From rjbuesa <@t> yahoo.com Thu Feb 9 12:36:45 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Feb 9 12:36:53 2006 Subject: [Histonet] Histonet controversies. Message-ID: <20060209183645.74495.qmail@web61219.mail.yahoo.com> Dear Colleagues: I only wish that answering to technical problems and queries would produce the same flurry of answers and advise. Most of all, so repetitive! Ren? J. --------------------------------- Brings words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. From rjbuesa <@t> yahoo.com Thu Feb 9 12:46:35 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Feb 9 12:46:44 2006 Subject: [Histonet] water for frozen section staining In-Reply-To: Message-ID: <20060209184635.88195.qmail@web61217.mail.yahoo.com> We always used running water at room temp. (sometimes quite "cold"). If you use warm/hot water maybe the section will "blue" sooner but just because the added effect of above room temp. of the water on the speed of the reaction. If temperature is going to affect the section adherence to the slide, it is probably the warm/hot water the one that could affect detrimentally the most. Ren? J. "Kapoor, Sue" wrote: Hello histonetters, We have a debate going in my lab as to using warm/hot or cold running water when staining frozen sections. One side states warm/hot water "blues" the hematoxylin...the other sides feels the cold water keeps the sections from falling off. What does everyone in histo land do and think??? thanks for your replies :) Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Brings words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. From MadaryJ <@t> MedImmune.com Thu Feb 9 13:15:50 2006 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Thu Feb 9 13:16:07 2006 Subject: [Histonet] aldehyde fuchsin prep and double stains for specials Message-ID: <7CAB706201F11843BD26AD516326F0C80EA102@MD1MS007.medimmune.com> Hello my heroes of the histonet, I am having trouble with the aldehyde fuchsin stain. Not the procedure, the reagent. I have made this stuff up more times than a headhunter whining. Twice I made it up, changed everything but the basic fuchsin each time. It will not ripen to that deep purple color, and will not work in procedure either. I checked the archives and see that one of the many masters of the microtome Johnny Kiernan recommends heating during the prep, I will try that. The paraldehyde was fresh both times, but was in the fridge at 4 degrees and froze because its melting point is like 11 degree or something. I called Sigma and they said it changes states all of the time in shipping and at the warehouse and freeze thaw on that stuff is insignificant. I never had a problem with that before anyway. Anyway, I need to do a Mast Cell stain and although the investigator is accepting TOL BLU she wants some different procedures, so I will do Torens, as well. But here is the other thing she wants a doubel stain for muci and mast cells, any ideas? I have tried some but will not share I want fresh thoughts. Nick Madary HT/HTL(ASCP)QIHC Histology Mgr, Medimmune Inc One Medimmune Way Gaithersburg, MD 20878 3013984745/6113 fax 9745 From gcallis <@t> montana.edu Thu Feb 9 13:23:51 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Feb 9 13:24:03 2006 Subject: [Histonet] Attention to those doing immunofluorescent staining, looking for mounting media Message-ID: <6.0.0.22.1.20060209121341.01b1a548@gemini.msu.montana.edu> Dear Histonetters doing immunofluorescence staining, Microscopy Today 14(1):34-39, January, 2006 just published Mounting Media and Antifade Reagents, Collins TJ This publication is a wonderful comparison of 10 different mounting medias for IFA work along with excellent discussion. I strongly recommend accessing and keeping a copy of this article around. It was not an endorsement, but certainly educational and useful. The only thing he did not include was using diluted (in it's own solvent) permanent mounting media (toluene or xylene based) instead of nail polish containing isopropyl alcohol as a sealant with nail polish (cited in literature) known to ruin GFP and cause it to not fluoresce. My thanks to Mr. Collins!! Enjoy Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From c.ingles <@t> hosp.wisc.edu Thu Feb 9 13:44:16 2006 From: c.ingles <@t> hosp.wisc.edu (Ingles Claire) Date: Thu Feb 9 13:48:59 2006 Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for yourhelp ...) References: Message-ID: <583D3E9A1E843445BD54E461D1A2F6F317A04F05@uwhis-xchng2.hosp.wisc.edu> Gee, and we're worried about postal workers? We are all trained in the use of sharp objects and have access to infectious agents... Think of the havoc a bunch of techs would wreak if we all went "histo". Maybe we all need to get out of the noxious fumes more often. Smile, spring is coming soon. (I think) Claire Ingles UW Mohs Clinic Madison WI ________________________________ From jkiernan <@t> uwo.ca Thu Feb 9 13:58:17 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Thu Feb 9 13:58:11 2006 Subject: [Histonet] aldehyde fuchsin prep and double stains for specials References: <7CAB706201F11843BD26AD516326F0C80EA102@MD1MS007.medimmune.com> Message-ID: <43EB9ED9.561E230F@uwo.ca> The basic fuchsine must be pararosaniline (CI 42500, Basic red 9). Other components of basic fuchsine (rosaniline, new fuchsine etc) won't work. You can get pararosaniline as a certified dye. The paraldehyde has to depolymerize. Hydrochloric acid is for the acid-catalyzed hydrolysis of paraldehyde, to generate acetaldehyde. Possibly old HCl might have lost some potency. It is, after all, a dissolved gas. The active substance released is acetaldehyde, which some people use directly. According to Peggy Wenk [J. Histotechnol. 19(4): 353, 1996] 2.5 ml of acetaldehyde can be substituted for 1.5 ml of paraldehyde). -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ "Madary, Joseph" wrote: > > Hello my heroes of the histonet, I am having trouble with the aldehyde fuchsin stain. Not the procedure, the reagent. I have made this stuff up more times than a headhunter whining. Twice I made it up, changed everything but the basic fuchsin each time. It will not ripen to that deep purple color, and will not work in procedure either. I checked the archives and see that one of the many masters of the microtome John Kiernan recommends heating during the prep, I will try that. The paraldehyde was fresh both times, but was in the fridge at 4 degrees and froze because its melting point is like 11 degree or something. I called Sigma and they said it changes states all of the time in shipping and at the warehouse and freeze thaw on that stuff is insignificant. I never had a problem with that before anyway. Anyway, I need to do a Mast Cell stain and although the investigator is accepting TOL BLU she wants some different procedures, so I will do Torens, as well. But here is the > other thing she wants a doubel stain for muci and mast cells, any ideas? I have tried some but will not share I want fresh thoughts. > > Nick Madary HT/HTL(ASCP)QIHC > Histology Mgr, Medimmune Inc > One Medimmune Way > Gaithersburg, MD 20878 > > 3013984745/6113 fax 9745 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Thu Feb 9 14:14:16 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Thu Feb 9 14:14:05 2006 Subject: [Histonet] B-Gal endogenous expression References: Message-ID: <43EBA298.B203A696@uwo.ca> Fairly recently: Dimri et 10 al (1995) Proc. Natl Acad. Sci. USA 92:9363-9367. More recently: Debacq-Chainlaux et 11 al (2005) J. Cell Sci. 118: 743-758. The lysosomal enzyme has a pH optimum about 4. Senescent cells contain a different enzyme active about pH 6. (The enzyme of the lac-Z reporter gene works at pH 7+ so it's not confused with the lysosomal enzyme activity.) In the major earlier papers on galactosidase histochemistry (1960s-'70s) the illustrations mainly showed intestine, with strong staining in epithelial cells; also renal tubules if I remember rightly. Hope this helps. John. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ "Andrea T. Hooper" wrote: > > Does anyone have references or anecdotal experience as to where > mammalian B-Gal is endogenously expressed if anywhere in mouse or > human? > > Thanks, > Andrea > -- From jkiernan <@t> uwo.ca Thu Feb 9 14:30:42 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Thu Feb 9 14:30:31 2006 Subject: [Histonet] mouse ovaries References: Message-ID: <43EBA672.1C172719@uwo.ca> Histological atlas of the laboratory mouse. William D. Gude, Gerald E. Cosgrove, and Gerald P. Hirsch. New York : Plenum Press, 1981. --- A Google search for mouse histology book quickly brought up an ad for this e-atlas: http://www.braintreesci.com/book14.htm --- A search in Google Images for mouse ovary histology brings up only 6 micrographs, of which three are not sections of ovary. The mouse ovary pictures weren't very good. It might be possible to find better, with more effort, but high quality micrographs of anything are not easy to find on the internet. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ "Edwards, R.E." wrote: > > Anyone know where I can find out about the histology of mouse ovaries??? > Many thanks > Richard Edwards > MRC TOXICOLOGY UNIT > LEICESTER...U.K...... > From KevinMcGovern <@t> catholichealth.net Thu Feb 9 14:54:15 2006 From: KevinMcGovern <@t> catholichealth.net (McGovern, Kevin) Date: Thu Feb 9 14:54:32 2006 Subject: [Histonet] RE: Claire Ingels' (re: Asking for your help ...) Message-ID: Claire-- Being a lowly BMET, I've been having a riot reading all these. I never worry about the techs here going Histo, as Xylene has such a CNS depressive effect! Besides, there's so much paraffin on the floor, you couldn't get enough traction to chase anyone down... :) Kevin -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Ingles Claire Sent: Thursday, February 09, 2006 1:44 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Public APOLOGY to the HistoNet (re: Asking foryourhelp ...) Gee, and we're worried about postal workers? We are all trained in the use of sharp objects and have access to infectious agents... Think of the havoc a bunch of techs would wreak if we all went "histo". Maybe we all need to get out of the noxious fumes more often. Smile, spring is coming soon. (I think) Claire Ingles UW Mohs Clinic Madison WI ________________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Thu Feb 9 14:58:03 2006 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Thu Feb 9 14:59:06 2006 Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for you r help ...) Message-ID: DITTO! What a LAME recruiter to think we're all so stupid. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of DDDeltour@mar.med.navy.mil Sent: Thu 2/9/2006 12:05 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Public APOLOGY to the HistoNet (re: Asking for you r help ...) Eric, If this were true then your original message would not have said..." Hi Histonetters". Admit that you used bad judgment and not accidentally sent the message to Histonet. Thanks -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Eric Dye Sent: Thursday, February 09, 2006 12:03 PM To: Histonetters Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for your help ...) Please accept my sincerest apologies for posting the email titled "Asking For Your Help" to the HistoNet. Those who commented are indeed correct: HistoNet is an inappropriate place for that type of posting. My list of HistoTechs is quite large. When a coworker and I built the list of prospective candidates, it was our intent to specifically exclude the HistoNet. Unfortunately, we goofed and the HistoNet got included. Once again accept my apology. Thank You for your understanding. Eric Dye, Sr HistoTech Recruiter PS: If you have any questions about this, call me at 800-466-9919 ext 223 ------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd Winter Park, FL 32792-6721 VOICE: 800-466-9919 ext 223 FAX: 407-671-6075 EMAIL: Eric@ategra.com ------------------------------------------------------- _______________________________________________ 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 ------------------------------- The information contained in this message may be privileged and / or confidential and protected from disclosure. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From glenn_krasinski <@t> yahoo.com Thu Feb 9 16:13:59 2006 From: glenn_krasinski <@t> yahoo.com (Glenn Krasinski) Date: Thu Feb 9 16:14:12 2006 Subject: [Histonet] A multi-use simple blocking solution?.... Message-ID: <20060209221359.67851.qmail@web37511.mail.mud.yahoo.com> I have been looking through the protocols on some of the IHC databases for blocking solutions. Some of the recipes are fairly exotic (fish skin gelatin?). Is there a good simple blocking solution that can be used in a variety of applications (primary/secondary)? How about 1x PBS with 1% BSA? Many thanks. Glenn M. Krasinski San Diego, CA --------------------------------- What are the most popular cars? Find out at Yahoo! Autos From mprice26 <@t> juno.com Thu Feb 9 16:34:19 2006 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Thu Feb 9 16:36:46 2006 Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for you rhel p ...) Message-ID: <20060209.143455.29586.235028@webmail65.nyc.untd.com> Histonetters (intentionally), HA! HA! HA! Not only is this listserve educational, it is extremely entertaining. We are a well rounded, unique group of people. That is why we have such a good time at our gatherings i.e. NSH etc. We are smart and love what we do, but also have a good sense of humor. Marsha Price From mprice26 <@t> juno.com Thu Feb 9 16:41:09 2006 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Thu Feb 9 16:42:50 2006 Subject: [Histonet] RE: Claire Ingels' (re: Asking for your help ...) Message-ID: <20060209.144154.29586.235078@webmail65.nyc.untd.com> Histonetters, I want everyone to vote on using Claire's phrase of "Go Histo" for the 2007 NSH Phrase. We could redo Prince's song to say "Let's go Histo" insted of "Let's Go Crazy". Marsha Price From tkngflght <@t> yahoo.com Thu Feb 9 16:44:47 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Thu Feb 9 16:45:56 2006 Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for yourhelp ...) In-Reply-To: <583D3E9A1E843445BD54E461D1A2F6F317A04F05@uwhis-xchng2.hosp.wisc.edu> Message-ID: <20060209224447.3096.qmail@web50906.mail.yahoo.com> I liked Kevin's response!! Some good came of all this--one of my best histo-friends from Seattle and I have been looking for each other---she emailed me this afternoon as a result of this exchange :) We need a T-Shirt---I've gone HISTO!!!! Cheryl Ingles Claire wrote: Gee, and we're worried about postal workers? We are all trained in the use of sharp objects and have access to infectious agents... Think of the havoc a bunch of techs would wreak if we all went "histo". Maybe we all need to get out of the noxious fumes more often. Smile, spring is coming soon. (I think) Claire Ingles UW Mohs Clinic Madison WI ________________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mprice26 <@t> juno.com Thu Feb 9 16:44:46 2006 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Thu Feb 9 16:46:54 2006 Subject: [Histonet] RE: "Let's Go Histo" motto for 2007 NSH Message-ID: <20060209.144458.29586.235099@webmail65.nyc.untd.com> Histonetters, All in favor of: "Let's Go Histo" for the 2007 NSH motto say yeah or nay. Marsha Price From gisela <@t> vetmed.wsu.edu Thu Feb 9 16:46:56 2006 From: gisela <@t> vetmed.wsu.edu (Bailey, Gisela) Date: Thu Feb 9 16:47:07 2006 Subject: [Histonet] Histology Manager Position Message-ID: <35CF12B690D8CA4E95375A36B4E7B44C0B6911@cvm36.vetmed.wsu.edu> NOTICE OF VACANCY WASHINGTON STATE UNIVERSITY College of Veterinary Medicine Washington Animal Disease Diagnostic Laboratory Tissue Processing Laboratory Manager - Search #4282 DESCRIPTION OF POSITION: The Tissue Processing Laboratory Manager is a full-time, permanent, administrative/professional position beginning approximately June 1, 2006, in the College of Veterinary Medicine, Washington Animal Disease Diagnostic Laboratory at Washington State University. MINIMUM QUALIFICATIONS: * Position requires a Bachelor's degree in a relevant field and four (4) years of progressively responsible experience in a field related to the functional managerial area which has included at least one (1) year of supervisory experience. Any combination of relevant education and experience may be substituted for educational requirement on a year-for-year basis. * ASCP Certification as a Histotechnician or Histotechnologist * Supervisory experience in a high volume, multiple-employee laboratory setting including the ability to anticipate and manage personnel issues, questions and conflicts effectively PREFERRED QUALIFICATIONS: * Experience in standard operating procedure (SOP) development and implementation for laboratory quality assurance and quality control practices * Experience trimming gross animal tissues for histologic evaluation * Familiarity with accredited laboratory procedures, including working in biosafety level 2 facilities * Experience with the coordination and implementation of laboratory health and environmental safety programs APPLICATION PROCESS: Review of applicants begins March 1, 2006. Submit a letter of interest addressing minimum and preferred qualifications, comprehensive r?sum?, and contact information for three professional references (include name, addresses, telephone numbers and email addresses) to: Ms. Myrle Cummings Washington Animal Disease Diagnostic Laboratory PO Box 647034 Pullman, WA 99164-7034 Fax: (509) 335-7424, E-mail: mlc@vetmed.wsu.edu SALARY: Starting salary $50,000 - $55,000/ year or commensurate with experience and education. MISC: Refer to www.vetmed.wsu.edu/depts_waddl for more information on the Washington Animal Disease Diagnostic Laboratory and the College of Veterinary Medicine. WASHINGTON STATE UNIVERSITY IS AN EQUAL OPPORTUNITY/AFFIRMATIVE ACTION EDUCATOR AND EMPLOYER. From jqb7 <@t> cdc.gov Thu Feb 9 18:10:18 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Thu Feb 9 18:08:46 2006 Subject: [Histonet] RE: "Let's Go Histo" motto for 2007 NSH References: <20060209.144458.29586.235099@webmail65.nyc.untd.com> Message-ID: Oh yeah! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of mprice26@juno.com Sent: Thu 2/9/2006 5:44 PM To: mprice26@juno.com Cc: pjfnefro@duke.edu; histonet@lists.utsouthwestern.edu; Terry.Marshall@rothgen.nhs.uk Subject: [Histonet] RE: "Let's Go Histo" motto for 2007 NSH Histonetters, All in favor of: "Let's Go Histo" for the 2007 NSH motto say yeah or nay. Marsha Price _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tkngflght <@t> yahoo.com Thu Feb 9 18:21:38 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Thu Feb 9 18:21:47 2006 Subject: [Histonet] "Let's Go Histo" motto for 2007 NSH In-Reply-To: Message-ID: <20060210002138.12323.qmail@web50914.mail.yahoo.com> Vote for "Let's Go Histo"--* NSH's contact info: phone: 301-262-6221 fax: 301-262-9188 e-mail: histo@nsh.org *Please take this posting with the humor with which it is intended!!! Don't call them or I'll be in hot water and they won't ever let me play in the sandbox again!!! Cheryl Histonetters, All in favor of: "Let's Go Histo" for the 2007 NSH motto say yeah or nay. Marsha Price From Linresearch <@t> aol.com Thu Feb 9 18:35:08 2006 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Thu Feb 9 18:35:26 2006 Subject: [Histonet] Perfusion set Message-ID: <1d8.4e7c09c7.311d39bc@aol.com> Hi, Can anyone recommend an automatic perfusion set? I need to perfuse rodents. Thanks, Lin From csr <@t> meyerinst.com Thu Feb 9 19:32:06 2006 From: csr <@t> meyerinst.com (Cheryl S. Rehfeld - Meyer Instruments, Inc.) Date: Thu Feb 9 19:32:22 2006 Subject: [Histonet] Louisiana Histology Meeting Message-ID: <001401c62de1$d5deae70$c79b210a@CherylWinbook> Relocated Histotechs, Due to hurricanes Katrina and Rita many of our Louisiana histotechs have been scattered far and wide. We are searching for you. Louisiana normally has its annual meeting in June but we don't know where you are now. If you would please respond to either Cheryl Crowder at ccrowder@vetmed.lsu.edu or myself at csr@meyerinst.com to let us know where you are currently located and if you would be interested in coming to the June meeting, then we would be able to better assess the situation. Also if you know where some of your fellow histotechs have gone we would love to know that as well. We miss you all and feel empty without you. Cheryl S. Rehfeld Meyer Instruments, Inc. C - 225-281-7739 O - 225-769-4465 F - 225-769-9885 E - csr@meyerinst.com From tamustuff <@t> yahoo.com Thu Feb 9 20:07:54 2006 From: tamustuff <@t> yahoo.com (Stephen Sperry) Date: Thu Feb 9 20:08:03 2006 Subject: [Histonet] new histo guy Message-ID: <20060210020754.16161.qmail@web37503.mail.mud.yahoo.com> My name is Stephen and I am an undergrad at Texas A&M Univ-Texarkana. I'm taking histology this spring semester and am really enjoying it so far. Any tips on how to survive an undergrad histo course? Thanks Stephen __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From ashokj <@t> glenmarkpharma.com Thu Feb 9 23:05:39 2006 From: ashokj <@t> glenmarkpharma.com (Ashok Jadhav) Date: Thu Feb 9 23:10:42 2006 Subject: [Histonet] RE: Histonet Digest, Vol 27, Issue 14 Message-ID: <2E1FBE2AC0130748B591FD48AC7056E30223A5D6@e2k1mhp.glenamrk.com> Dear all Histonet subscriber I want to stain the eye tissue of rat that were fixed in Zenker's solution and paraffine embedded with normal processing but I am not able to stain with HE stain's. Could Anyone tell me the which method should I follow for the staining of eye tissue. AShok DISCLAIMER: The information contained in this email communication is intended only for the personal and confidential use of the designated recipient named above. This message may be an attorney-client communication, and as such is privileged and confidential. If the reader of this message is not the intended recipient, you are hereby notified that you have received this communication in error, and that any review, dissemination, distribution, or copying of the message is strictly prohibited. If you have received this transmission in error, please destroy this transmission and notify Glenmark Pharmaceuticals Limited immediately by telephone and/or send an email to supportho@glenmarkpharma.com From louise.renton <@t> gmail.com Fri Feb 10 00:47:34 2006 From: louise.renton <@t> gmail.com (louise renton) Date: Fri Feb 10 00:47:45 2006 Subject: [Histonet] RE: Ihc bone staining In-Reply-To: References: Message-ID: Rhonda depends on what you are looking for, perhaps the expression in bone is very low and thus is not as apparent as in other cells. Certain of the bone morphogens eg BMP3 are also expressed in Purkinje cells, as I am sure are many other antibodies. Perhaps a little more detail as to what yoyu are specifically looking at/for would be helpful best regards On 2/9/06, Allen, Rhonda wrote: > > > > Does anyone know if it could possibly take longer(or more concentrated > > antibody) to stain the bone in a tissue section(mouse head, newborn > > pup, not decalcified), vs. the other tissue components(brain, skin). > > In order to get nice staining in the bone, seems like we are starting > > to see background coming up in the brain. Wondering if this is normal > > for some antibodies. > > Thanks in advance, > > Rhonda Allen BA HT(ASCP)HTL, QIHC > > Histotechnology Specialist II > > Stowers Institute > > 1000 E. 50th Street > > Kansas City, MO 64110 > > 816-926-4305 > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "Does a conceited cowboy, only ride on pompous grass?. From kemlo.rogerson <@t> waht.swest.nhs.uk Fri Feb 10 03:28:19 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Fri Feb 10 03:28:43 2006 Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for you r help ...) Message-ID: Poor guys wearing a 'hair shirt'. Him bad boy but stop the feeding frenzy!! Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Linke_Noelle [mailto:Linke_Noelle@Allergan.com] Sent: Thursday, February 09, 2006 5:13 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Public APOLOGY to the HistoNet (re: Asking for your help ...) You people need to chill out and get over this....... If you don't like what you read, the 'delete' key is right there for you to use..... Who are you to judge??? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DDDeltour@mar.med.navy.mil Sent: Thursday, February 09, 2006 9:05 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Public APOLOGY to the HistoNet (re: Asking for your help ...) Eric, If this were true then your original message would not have said..." Hi Histonetters". Admit that you used bad judgment and not accidentally sent the message to Histonet. Thanks -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Eric Dye Sent: Thursday, February 09, 2006 12:03 PM To: Histonetters Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for your help ...) Please accept my sincerest apologies for posting the email titled "Asking For Your Help" to the HistoNet. Those who commented are indeed correct: HistoNet is an inappropriate place for that type of posting. My list of HistoTechs is quite large. When a coworker and I built the list of prospective candidates, it was our intent to specifically exclude the HistoNet. Unfortunately, we goofed and the HistoNet got included. Once again accept my apology. Thank You for your understanding. Eric Dye, Sr HistoTech Recruiter PS: If you have any questions about this, call me at 800-466-9919 ext 223 ------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd Winter Park, FL 32792-6721 VOICE: 800-466-9919 ext 223 FAX: 407-671-6075 EMAIL: Eric@ategra.com ------------------------------------------------------- _______________________________________________ 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 From kemlo.rogerson <@t> waht.swest.nhs.uk Fri Feb 10 03:31:13 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Fri Feb 10 03:31:29 2006 Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for you rhelp ...) Message-ID: Maybe disembowelling? Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Thursday, February 09, 2006 5:46 PM To: Pat Flannery; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Public APOLOGY to the HistoNet (re: Asking for you rhelp ...) I agree to the harsh bit. Two have been "offended" however, so perhaps we should organise a public march, with banners demanding that the perpetrator be beheaded. There is a historical precedence. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Pat Flannery [mailto:pjfnefro@duke.edu] Sent: 09 February 2006 17:26 To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Public APOLOGY to the HistoNet (re: Asking for you rhelp ...) Gee, let's not only reject his apology, let's publicly flog the guy. Boy, you're pretty harsh on this list! -Pat Flannery pjfnefro@duke.edu On Feb 9, 2006, at 12:05 PM, DDDeltour@mar.med.navy.mil wrote: > Eric, > If this were true then your original message would not have > said..." Hi > Histonetters". Admit that you used bad judgment and not > accidentally sent > the message to Histonet. Thanks > > _______________________________________________ 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 From ree3 <@t> leicester.ac.uk Fri Feb 10 03:43:51 2006 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Feb 10 03:44:03 2006 Subject: [Histonet] RE: Claire Ingels' (re: Asking for your help ...) Message-ID: or Go to HistoHeaven in 2007??? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of mprice26@juno.com Sent: 09 February 2006 22:41 To: KevinMcGovern@catholichealth.net Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: Claire Ingels' (re: Asking for your help ...) Histonetters, I want everyone to vote on using Claire's phrase of "Go Histo" for the 2007 NSH Phrase. We could redo Prince's song to say "Let's go Histo" insted of "Let's Go Crazy". Marsha Price _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Fri Feb 10 06:22:30 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Fri Feb 10 06:22:35 2006 Subject: [Histonet] Subject line Message-ID: May I respectfully suggest that if you are sending an email to Histonet that you not use an old subject line that has nothing to do with what you have in the text of your message. I would really appeciate this as it would make our use of Histonet and deletion of items of non interest a lot easier. Thank y'all and have agreat weekend. Barry Barry From codex_code <@t> yahoo.com Fri Feb 10 06:29:44 2006 From: codex_code <@t> yahoo.com (Marie Hoh) Date: Fri Feb 10 06:29:52 2006 Subject: [Histonet] Information on NZ Message-ID: <20060210122944.1676.qmail@web37612.mail.mud.yahoo.com> Dear all, I have a Bachelor's degree in Biological Sciences and I have been working in a research institution for 3 years. My job consists mainly of immunohistochemistry, histology and molecular biology work involving techniques such as PCR and gel electrophoresis. I am looking for employment in hospitals or research centers in New Zealand. I would be much obliged if you all could help me in how to go about this. Could you kindly email me at codex_code@yahoo.com? Your help would be much appreciated. Thanks, Marie __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From gpbnas <@t> yahoo.es Fri Feb 10 08:11:55 2006 From: gpbnas <@t> yahoo.es (Guillermo Palao) Date: Fri Feb 10 08:12:05 2006 Subject: [Histonet] Mouse tissue staining with monoclonal mouse Abs. Help with Vector Mouse on Mouse kit. Message-ID: <20060210141155.23662.qmail@web26203.mail.ukl.yahoo.com> Dear all, I am working with mouse frozen sections of different autoimmune disease models, implying an important mouse IgG deposition in studied tissues (mouse kidney). For this reason we bought Vector Mouse on Mouse kit to avoid detection of deposited IgG by secondary antibodies while maintaining sensitivity for mouse primary antibody. After increasing concentration of blocking reagent as well as decreasing concentration of biotynilated anti-mouse IgG, there still is a "background" of positive staining of deposited IgG in renal glomeruli together with specific staining of antigens of interest. I would really appreciate any tips or ideas you might think helpful. Thanks in advance, Guillermo Palao, MD. Ph.D. Laboratorio de Reumatolog?a Centro de Investigaci?n Hospital 12 de Octubre Avda de C?rdoba s/n Madrid 28041 Spain --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com From vazquezr <@t> ohsu.edu Fri Feb 10 09:12:28 2006 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Feb 10 09:12:55 2006 Subject: [Histonet] Public APOLOGY to the HistoNet (re: Asking for you r help ...) Message-ID: I don't buy it either, even if it was on sale. Hi Histonetter is directed staight at us. Robyn From Janet.Bonner <@t> FLHOSP.ORG Fri Feb 10 09:20:37 2006 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Feb 10 09:21:21 2006 Subject: [Histonet] RE: "Let's Go Histo" motto for 2007 NSH Message-ID: YEA!!!!!!! ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of mprice26@juno.com Sent: Thu 2/9/2006 5:44 PM To: mprice26@juno.com Cc: pjfnefro@duke.edu; histonet@lists.utsouthwestern.edu; Terry.Marshall@rothgen.nhs.uk Subject: [Histonet] RE: "Let's Go Histo" motto for 2007 NSH Histonetters, All in favor of: "Let's Go Histo" for the 2007 NSH motto say yeah or nay. Marsha Price _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------- The information contained in this message may be privileged and / or confidential and protected from disclosure. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From gcallis <@t> montana.edu Fri Feb 10 09:45:50 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Feb 10 09:46:01 2006 Subject: [Histonet] A multi-use simple blocking solution?.... In-Reply-To: <20060209221359.67851.qmail@web37511.mail.mud.yahoo.com> References: <20060209221359.67851.qmail@web37511.mail.mud.yahoo.com> Message-ID: <6.0.0.22.1.20060210084254.01b48bb0@gemini.msu.montana.edu> We prefer to use normal serum blocking, with the serum matched to the host of the secondary and do not use BSA based blockers. This has been discussed at great lengths on Histonet, check out Histonet Archives. Blocking, per your question, is done many ways. I suggest you do to the DAKO website, information and access the Handbook of Immunochemical Methods, Boesnish discusses various blocking agents, and you can download this wonderful freebie in pdf format. You can buy superblocks ready to use from various companies, i.e.At 03:13 PM 2/9/2006, you wrote: >I have been looking through the protocols on some of the IHC databases for >blocking solutions. Some of the recipes are fairly exotic (fish skin >gelatin?). Is there a good simple blocking solution that can be used in a >variety of applications (primary/secondary)? How about 1x PBS with 1% BSA? > > Many thanks. > > Glenn M. Krasinski > San Diego, CA > > >--------------------------------- > > What are the most popular cars? Find out at Yahoo! Autos >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From funderwood <@t> mcohio.org Fri Feb 10 09:51:08 2006 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Feb 10 09:51:36 2006 Subject: [Histonet] RE: Claire Ingels' (re: Asking for your help ...) Message-ID: Histo-ry of the World >>> "Edwards, R.E." 02/10/06 04:43AM >>> or Go to HistoHeaven in 2007??? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of mprice26@juno.com Sent: 09 February 2006 22:41 To: KevinMcGovern@catholichealth.net Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: Claire Ingels' (re: Asking for your help ...) Histonetters, I want everyone to vote on using Claire's phrase of "Go Histo" for the 2007 NSH Phrase. We could redo Prince's song to say "Let's go Histo" insted of "Let's Go Crazy". Marsha Price _______________________________________________ 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 From fmonson <@t> wcupa.edu Fri Feb 10 09:54:38 2006 From: fmonson <@t> wcupa.edu (Monson, Frederick ) Date: Fri Feb 10 09:54:50 2006 Subject: [Histonet] RE: Claire Ingels' (re: Asking for your help...) Message-ID: Is this going to end up as, "Those who don't learn their Histo are doomed to repeat it?" Frederick C. Monson, PhD Technical Director Light, Electron, X-Ray and Scanning Probe Imaging and Analysis Center (LEXSPIAC) Large Scientific Instrument Core Geology, West Chester University S. Church St. and W. Rosedale Ave. West Chester, PA, 19320 610-738-0437 fmonson@wcupa.edu URL: http://darwin.wcupa.edu/cgi-bin/casireserve.cgi ============================================ Knowledge is the key to happiness. Ignorance might be the key to contentment. ============================================ -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred Underwood Sent: Friday, February 10, 2006 10:51 AM To: KevinMcGovern@catholichealth.net; mprice26@juno.com; ree3@leicester.ac.uk Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Claire Ingels' (re: Asking for your help...) Histo-ry of the World >>> "Edwards, R.E." 02/10/06 04:43AM >>> or Go to HistoHeaven in 2007??? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of mprice26@juno.com Sent: 09 February 2006 22:41 To: KevinMcGovern@catholichealth.net Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: Claire Ingels' (re: Asking for your help ...) Histonetters, I want everyone to vote on using Claire's phrase of "Go Histo" for the 2007 NSH Phrase. We could redo Prince's song to say "Let's go Histo" insted of "Let's Go Crazy". Marsha Price _______________________________________________ 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 From gcallis <@t> montana.edu Fri Feb 10 09:59:11 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Feb 10 09:59:23 2006 Subject: [Histonet] "Let's Go Histo" motto for 2007 NSH In-Reply-To: <20060210002138.12323.qmail@web50914.mail.yahoo.com> References: <20060210002138.12323.qmail@web50914.mail.yahoo.com> Message-ID: <6.0.0.22.1.20060210085649.01b60aa8@gemini.msu.montana.edu> I think you will find NSH has already selected a motto for Phoenix meeting and did so well a long time ago. The 2006 NSH S/C motto is Rising to Greater Heights. Maybe Histonet should have this motto instead!??? At 05:21 PM 2/9/2006, you wrote: >Vote for "Let's Go Histo"--* > > NSH's contact info: > phone: 301-262-6221 >fax: 301-262-9188 >e-mail: histo@nsh.org > > > Cheryl Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From gcallis <@t> montana.edu Fri Feb 10 10:00:11 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Feb 10 10:00:23 2006 Subject: [Histonet] new histo guy In-Reply-To: <20060210020754.16161.qmail@web37503.mail.mud.yahoo.com> References: <20060210020754.16161.qmail@web37503.mail.mud.yahoo.com> Message-ID: <6.0.0.22.1.20060210085938.01b7ed78@gemini.msu.montana.edu> Study and read!! At 07:07 PM 2/9/2006, you wrote: >My name is Stephen and I am an undergrad at Texas A&M Univ-Texarkana. I'm >taking histology this spring semester and am really enjoying it so far. >Any tips on how to survive an undergrad histo course? Thanks > > Stephen Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From info <@t> instrumedics.com Fri Feb 10 10:01:03 2006 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Feb 10 10:01:23 2006 Subject: [Histonet] mounting media Message-ID: <00c201c62e5b$389be400$6401a8c0@INSTRUMEDICS22> I read the interesting review of mounting media by Tony Collins in Microscopy Today. I regret that CureMount, which is an Instrumedics product was not evaluated. was developed to have a refractive index that matches that of dehydrated and cleared frozen and paraffin sections. It contains a photocatalyst which makes it possible to "cure" the CureMount with 20 sec UV exposure . The slide is "dry" and can actually be filed immedicatedly. The best part is that the optical properties in the light microscope make the background virtually invisible and the stained structures have greater clarity. Bernice From pmarcum <@t> vet.upenn.edu Fri Feb 10 10:13:43 2006 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Fri Feb 10 10:14:15 2006 Subject: [Histonet] RE: Claire Ingels' (re: Asking for your help...) In-Reply-To: References: Message-ID: <6.1.1.1.2.20060210110439.01945ec0@mail.vet.upenn.edu> I was under the impression the state holding the meeting made this decision not the NSH office. Therefore you should be talking to or directing this to Dallas and/or the Texas group if that is where the meeting is for 07. NSH handles the main meeting and the state/region are responsible for the theme of the meeting and the banquet functions as well as some of the PR. The state/region also are instrumental in supplying and rounding up volunteers for all the behind the scenes work required to make attendees feel it is seamless. NSH committee people assure this goes smoothly. As Pennsylvania is holding the 08 meeting I have been clarifying what we will be required to do and that is the only reason I know about this. We should all find out what the states and regions are responsible for at the National meeting so we have a better idea where and how we can help our local organizations have the best meetings possible and represent ourselves better. Pam Marcum At 10:54 AM 2/10/2006, Monson, Frederick wrote: >Is this going to end up as, "Those who don't learn their Histo are >doomed to repeat it?" > >Frederick C. Monson, PhD >Technical Director >Light, Electron, X-Ray and Scanning Probe Imaging and Analysis Center >(LEXSPIAC) >Large Scientific Instrument Core >Geology, West Chester University >S. Church St. and W. Rosedale Ave. >West Chester, PA, 19320 >610-738-0437 >fmonson@wcupa.edu >URL: http://darwin.wcupa.edu/cgi-bin/casireserve.cgi > >============================================ >Knowledge is the key to happiness. Ignorance might be the key to >contentment. >============================================ > > > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred >Underwood >Sent: Friday, February 10, 2006 10:51 AM >To: KevinMcGovern@catholichealth.net; mprice26@juno.com; >ree3@leicester.ac.uk >Cc: histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] RE: Claire Ingels' (re: Asking for your help...) > >Histo-ry of the World > > >>> "Edwards, R.E." 02/10/06 04:43AM >>> >or Go to HistoHeaven in 2007??? > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of >mprice26@juno.com >Sent: 09 February 2006 22:41 >To: KevinMcGovern@catholichealth.net >Cc: histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] RE: Claire Ingels' (re: Asking for your help >...) > > >Histonetters, >I want everyone to vote on using Claire's phrase of "Go Histo" for the >2007 NSH Phrase. We could redo Prince's song to say "Let's go Histo" >insted of "Let's Go Crazy". > >Marsha Price > > >_______________________________________________ >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 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From froyer <@t> bitstream.net Fri Feb 10 10:15:04 2006 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Feb 10 10:15:22 2006 Subject: [Histonet] "Let's Go Histo" motto for 2007 NSH In-Reply-To: <6.0.0.22.1.20060210085649.01b60aa8@gemini.msu.montana.edu> Message-ID: <007e01c62e5d$275620f0$6f01a80a@fords> Now Gayle; don't go HISTO on us... ;-) ~ Ford :-) Ford M. Royer, MT(ASCP) Minnesota Medical, Inc. Minneapolis, MN 55427 888-790-9686 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Friday, February 10, 2006 9:59 AM To: Cheryl; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] "Let's Go Histo" motto for 2007 NSH I think you will find NSH has already selected a motto for Phoenix meeting and did so well a long time ago. The 2006 NSH S/C motto is Rising to Greater Heights. Maybe Histonet should have this motto instead!??? At 05:21 PM 2/9/2006, you wrote: >Vote for "Let's Go Histo"--* > > NSH's contact info: > phone: 301-262-6221 >fax: 301-262-9188 >e-mail: histo@nsh.org > > > Cheryl Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hborgeri <@t> wfubmc.edu Fri Feb 10 10:20:30 2006 From: hborgeri <@t> wfubmc.edu (Hermina Borgerink) Date: Fri Feb 10 10:24:10 2006 Subject: [Histonet] A multi-use simple blocking solution?.... Message-ID: <9AEEF1FB6254224AA355ED285F8491651675BEEE@EXCHVS2.medctr.ad.wfubmc.edu> 0.5% casein in your wash buffer and diluents. Ref.: Tacha, D and McKinney, L. Casein Reduces Nonspecific Background staining in Immunolabeling Techniques. J Histotechnology 15: 2, June 1992 Hermina -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Friday, February 10, 2006 10:46 AM To: Glenn Krasinski; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] A multi-use simple blocking solution?.... We prefer to use normal serum blocking, with the serum matched to the host of the secondary and do not use BSA based blockers. This has been discussed at great lengths on Histonet, check out Histonet Archives. Blocking, per your question, is done many ways. I suggest you do to the DAKO website, information and access the Handbook of Immunochemical Methods, Boesnish discusses various blocking agents, and you can download this wonderful freebie in pdf format. You can buy superblocks ready to use from various companies, i.e.At 03:13 PM 2/9/2006, you wrote: >I have been looking through the protocols on some of the IHC databases >for blocking solutions. Some of the recipes are fairly exotic (fish >skin gelatin?). Is there a good simple blocking solution that can be >used in a variety of applications (primary/secondary)? How about 1x PBS with 1% BSA? > > Many thanks. > > Glenn M. Krasinski > San Diego, CA > > >--------------------------------- > > What are the most popular cars? Find out at Yahoo! Autos >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BNuern <@t> coj.net Fri Feb 10 10:34:16 2006 From: BNuern <@t> coj.net (Barb Nuernberger) Date: Fri Feb 10 10:34:44 2006 Subject: [Histonet] Does anyone have any suggestions for getting brain tissue (human) to stay on the slides? Message-ID: Does anyone have any suggestions for getting brain tissue (human) to stay on the slides? Fixation? Barb From rjbuesa <@t> yahoo.com Fri Feb 10 10:41:18 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Feb 10 10:41:27 2006 Subject: [Histonet] Does anyone have any suggestions for getting brain tissue (human) to stay on the slides? In-Reply-To: Message-ID: <20060210164118.46138.qmail@web61212.mail.yahoo.com> Usually is a problem with processing, or too thick a section, or not "dried" (heated) enough after section or a combination. Ren? J. Barb Nuernberger wrote: Does anyone have any suggestions for getting brain tissue (human) to stay on the slides? Fixation? Barb _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Brings words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. From tkngflght <@t> yahoo.com Fri Feb 10 10:42:07 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Fri Feb 10 10:42:15 2006 Subject: [Histonet] NSH Meetings- In-Reply-To: <6.0.0.22.1.20060210085649.01b60aa8@gemini.msu.montana.edu> Message-ID: <20060210164207.90780.qmail@web50908.mail.yahoo.com> I assume there are lots of volunteer opportunities for each national NSH symposium. Would the people who run the next few events post a contact? I would like to help and know a few others who can do the same--thanks in advance!! Cheryl Full Staff Inc Gayle Callis wrote: I think you will find NSH has already selected a motto for Phoenix meeting and did so well a long time ago. The 2006 NSH S/C motto is Rising to Greater Heights. Maybe Histonet should have this motto instead!??? At 05:21 PM 2/9/2006, you wrote: >Vote for "Let's Go Histo"--* > > NSH's contact info: > phone: 301-262-6221 >fax: 301-262-9188 >e-mail: histo@nsh.org > > > Cheryl Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From tkngflght <@t> yahoo.com Fri Feb 10 10:51:45 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Fri Feb 10 10:51:54 2006 Subject: [Histonet] brain tissue (human) to stay on the slides? In-Reply-To: <20060210164118.46138.qmail@web61212.mail.yahoo.com> Message-ID: <20060210165145.66511.qmail@web50907.mail.yahoo.com> I know of no scientific explanation to this method but from experience, you usually have to scrape the tissue off with a blade if you want it to come off afterward: Double Baking: Oven dry your slides for the usual time. Pull them out and let them cool COMPLETELY. Then put them back in the same oven for the same amount of time, and cool completely again. It seems to 'settle' the tissue against the slides by heating and cooling twice. This works for most soft tissues. Decal and nail sections didn't work as completely. This will effect IHC but works beautifully with no stain changes for H&E and most specials. Also--always use treated slides, and watch the speed of your rinse waters/aggitation. Hope this helps!! Cheryl Kerry Full Staff Inc. Staffing the AP Lab by helping one tech at a time. Rene J Buesa wrote: Usually is a problem with processing, or too thick a section, or not "dried" (heated) enough after section or a combination. Ren? J. Barb Nuernberger wrote: Does anyone have any suggestions for getting brain tissue (human) to stay on the slides? Fixation? Barb _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Brings words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mprice26 <@t> juno.com Fri Feb 10 10:51:19 2006 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Feb 10 10:52:26 2006 Subject: [Histonet] "Let's Go Histo" motto for 2007 NSH Message-ID: <20060210.085120.12386.93095@webmail45.nyc.untd.com> I intended this for humor only. Although is is a catchy phrase. I know that NSH chooses there theme way ahead of time. Just trying to get everybody to have a little Histo-Humor. Marsha From funderwood <@t> mcohio.org Fri Feb 10 10:55:51 2006 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Feb 10 10:56:21 2006 Subject: [Histonet] "Let's Go Histo" motto for 2007 NSH Message-ID: Why it's Hist-erical. >>> "mprice26@juno.com" 02/10/06 11:51AM >>> I intended this for humor only. Although is is a catchy phrase. I know that NSH chooses there theme way ahead of time. Just trying to get everybody to have a little Histo-Humor. Marsha _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Fri Feb 10 10:58:52 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Fri Feb 10 10:58:57 2006 Subject: [Histonet] new histo guy Message-ID: Stephen I would read the relevant chapter before attending that lecture on the topic. I don't know which text you have been assigned there are several out there. Some suggestions for books that help when you are a beginner. National Medical series for Independent Study. Histology. Ray C. Henrikson, Gordon I. Kaye and Joseph E. Mazurkiewicz. 1997. Published by Lippincott Williams and Wilkins. A Learning System in Histology. CD Rom and Guide. Deborah W. Vaughan. Oxford press. 2002. We use Basic Histology a Text and Atlas. 11th edition. 2005. Luiz Junqueira and Jose Carneiro. McGraw Hill. Comes with a CD ROM. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Friday, February 10, 2006 10:00 AM To: Stephen Sperry; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] new histo guy Study and read!! At 07:07 PM 2/9/2006, you wrote: >My name is Stephen and I am an undergrad at Texas A&M Univ-Texarkana. I'm >taking histology this spring semester and am really enjoying it so far. >Any tips on how to survive an undergrad histo course? Thanks > > Stephen Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mprice26 <@t> juno.com Fri Feb 10 10:57:40 2006 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Feb 10 10:59:15 2006 Subject: [Histonet] "Let's Go Histo" motto for 2007 NSH Message-ID: <20060210.085828.12386.93155@webmail45.nyc.untd.com> HA! HA! See we histotechs do have a sense of humor. Marsha From dellav <@t> musc.edu Fri Feb 10 11:00:16 2006 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Feb 10 11:02:36 2006 Subject: [Histonet] NSH Meetings- Message-ID: Anyone interested in helping at an NSH Symposium may contact Aubrey Wanner, NSH meeting manager, at aubrey@nsh.org also, conference mottos are typically recommended to NSH by the local state committee that is hosting the conference. of course, there is no reason why histonetters could not make recommendations to the state society. the 07 conference will be in Denver Colorado. contact the Colorado society if you have suggestions for motto (NOT the NSH office !) Dallas was originally announced as the 07 conference site. The NSH board of directors recently voted to move the conference from Dallas because the space originally contracted for was insufficient to meet our current needs. Not to bore you with the details, but at one time we contracted 7-10 years out to increase our likelihood of getting the locations we wanted. however, growth in attendance at our conference over the past few years makes it impractical to plan that far ahead. we wouldnt want to have to turn any histotechs or vendors away who want to attend simply because we didn't have the space to accommodate them. Vinnie >>> Cheryl 02/10/06 11:42AM >>> I assume there are lots of volunteer opportunities for each national NSH symposium. Would the people who run the next few events post a contact? I would like to help and know a few others who can do the same--thanks in advance!! Cheryl Full Staff Inc Gayle Callis wrote: I think you will find NSH has already selected a motto for Phoenix meeting and did so well a long time ago. The 2006 NSH S/C motto is Rising to Greater Heights. Maybe Histonet should have this motto instead!??? At 05:21 PM 2/9/2006, you wrote: >Vote for "Let's Go Histo"--* > > NSH's contact info: > phone: 301-262-6221 >fax: 301-262-9188 >e-mail: histo@nsh.org > > > Cheryl Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RohrT <@t> nyackhospital.org Fri Feb 10 11:03:10 2006 From: RohrT <@t> nyackhospital.org (Theresa Rohr) Date: Fri Feb 10 11:04:54 2006 Subject: [Histonet] Labvision Neomarkers GCDFP-15 Ab1, Clone 23A3 Message-ID: Dear Histonetters, I am using Neomarker's GCDFP Ab-1, Clone 23A3 at a 1:25 titer. I use a 6.0 citrate for Target Retrieval for 20 minutes in a Steamer and 20 minute cooling. I stain on the Dako Autostainer with Envision+ at 30 minutes in Primary Ab, 30 minutes in Polymer and 5 minutes in DAB. The staining results are just OK nothing extraordinary. Has anyone used this antibody and had great results? If so, what are you doing differently than my protocol? Do you think a 9.0 EDTA buffer for Retrieval would be better? What about increasing my time in DAB? Any help would be greatly appreciated. I have emailed their techservice but thought my peers might have more to teach me. Thanks so much, Theresa Rohr rohrt@nyackhospital.org Theresa Rohr, BA HT(ASCP) Section Head, Histology Nyack Hospital 160 North Midland Avenue Nyack, New York 10960 phone - 845-348-2276 fax - 845-348-8430 Nyack Hospital is required by Law to protect the privacy of health information under HIPAA Rules and Regulations Part 11, Article 45 CFR, Parts 160-164. This email, including attachments, is intended for the exclusive use of the person or the entity to which it is addressed. It may contain confidential or legally protected information. The authorized recipient is prohibited from disclosing this information to any other party and is required to destroy this information after its stated need has been fulfilled. If the recipient or reader of this email is not the intended recipient or her/his authorized agent, the reader is hereby notified that any dissemination, distribution or copying of this email is phohibited. If you believe that you have received this email in error, please advise the sender immediately by reply email and then delete this email immediately. From tkngflght <@t> yahoo.com Fri Feb 10 11:08:33 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Fri Feb 10 11:08:42 2006 Subject: [Histonet] NSH Meetings-contacts and venues In-Reply-To: Message-ID: <20060210170833.52388.qmail@web50914.mail.yahoo.com> Thanks everyone! Ms. Marcum suggested that to offer assistance, keep up with our science as well as the venues and contacts for all up and coming Symposioums at the national level we utilize the www.nsh.org web site. As she aptly points out: it is the best place to go for information in all areas of our society... besides here, of course!! Happy Friday!! Cheryl Full Staff Inc. Staffing the AP Lab by helping one tech at a time. Vinnie Della Speranza wrote: Anyone interested in helping at an NSH Symposium may contact Aubrey Wanner, NSH meeting manager, at aubrey@nsh.org From dellav <@t> musc.edu Fri Feb 10 11:05:29 2006 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Feb 10 11:11:15 2006 Subject: [Histonet] new histo guy Message-ID: Stephen, good luck to you. I hope that you find that histology becomes your passion. in addition to the good advice offered by Barry and Gayle, I'd add that I occasionally run into students who seriously underestimated the effort required to master this subject. It is impossible not to be awed by the complexity of the human anatomy and physiology. if you do not devote sufficient time to reading and studying you will not be successful in this subject matter. Histology is our passion. Let it be yours by giving yourself the best chance to master it. Vinnie >>> "Rittman, Barry R" 02/10/06 11:58AM >>> Stephen I would read the relevant chapter before attending that lecture on the topic. I don't know which text you have been assigned there are several out there. Some suggestions for books that help when you are a beginner. National Medical series for Independent Study. Histology. Ray C. Henrikson, Gordon I. Kaye and Joseph E. Mazurkiewicz. 1997. Published by Lippincott Williams and Wilkins. A Learning System in Histology. CD Rom and Guide. Deborah W. Vaughan. Oxford press. 2002. We use Basic Histology a Text and Atlas. 11th edition. 2005. Luiz Junqueira and Jose Carneiro. McGraw Hill. Comes with a CD ROM. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Friday, February 10, 2006 10:00 AM To: Stephen Sperry; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] new histo guy Study and read!! At 07:07 PM 2/9/2006, you wrote: >My name is Stephen and I am an undergrad at Texas A&M Univ-Texarkana. I'm >taking histology this spring semester and am really enjoying it so far. >Any tips on how to survive an undergrad histo course? Thanks > > Stephen Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 _______________________________________________ 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 From pruegg <@t> ihctech.net Fri Feb 10 11:24:17 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Feb 10 11:24:25 2006 Subject: [Histonet] NSH Meetings- In-Reply-To: Message-ID: <200602101724.k1AHOD3t023833@chip.viawest.net> Anyone wishing to help with the NSH S/C now being held in Denver in 2007 can contact me. We need your help. Colorado and Region VII are very small in numbers of histotechs, although very large in area. Having the S/C in Region VII in 2006 and 2007 will certainly test the abilities of the locals to pull this off, so any help we can solicit even outside Region VII is welcomed. Thank you and welcome to Colorado in 2007, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vinnie Della Speranza Sent: Friday, February 10, 2006 10:00 AM To: Histonet@lists.utsouthwestern.edu; gcallis@montana.edu; tkngflght@yahoo.com Subject: Re: [Histonet] NSH Meetings- Anyone interested in helping at an NSH Symposium may contact Aubrey Wanner, NSH meeting manager, at aubrey@nsh.org also, conference mottos are typically recommended to NSH by the local state committee that is hosting the conference. of course, there is no reason why histonetters could not make recommendations to the state society. the 07 conference will be in Denver Colorado. contact the Colorado society if you have suggestions for motto (NOT the NSH office !) Dallas was originally announced as the 07 conference site. The NSH board of directors recently voted to move the conference from Dallas because the space originally contracted for was insufficient to meet our current needs. Not to bore you with the details, but at one time we contracted 7-10 years out to increase our likelihood of getting the locations we wanted. however, growth in attendance at our conference over the past few years makes it impractical to plan that far ahead. we wouldnt want to have to turn any histotechs or vendors away who want to attend simply because we didn't have the space to accommodate them. Vinnie >>> Cheryl 02/10/06 11:42AM >>> I assume there are lots of volunteer opportunities for each national NSH symposium. Would the people who run the next few events post a contact? I would like to help and know a few others who can do the same--thanks in advance!! Cheryl Full Staff Inc Gayle Callis wrote: I think you will find NSH has already selected a motto for Phoenix meeting and did so well a long time ago. The 2006 NSH S/C motto is Rising to Greater Heights. Maybe Histonet should have this motto instead!??? At 05:21 PM 2/9/2006, you wrote: >Vote for "Let's Go Histo"--* > > NSH's contact info: > phone: 301-262-6221 >fax: 301-262-9188 >e-mail: histo@nsh.org > > > Cheryl Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ 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 From tkngflght <@t> yahoo.com Fri Feb 10 10:38:24 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Fri Feb 10 11:31:45 2006 Subject: [Histonet] mounting media In-Reply-To: <00c201c62e5b$389be400$6401a8c0@INSTRUMEDICS22> Message-ID: <20060210163824.2181.qmail@web50903.mail.yahoo.com> I'd like to learn more about this-- Is this used with coverslips or like flo-tex (sp?) is used--poured over a slide? What are the storage statistics (temp/humidity/longevity) and can we try it without breaking the budget or a patented UV source?? Instrumedics wrote: I read the interesting review of mounting media by Tony Collins in Microscopy Today. I regret that CureMount, which is an Instrumedics product was not evaluated. was developed to have a refractive index that matches that of dehydrated and cleared frozen and paraffin sections. It contains a photocatalyst which makes it possible to "cure" the CureMount with 20 sec UV exposure . The slide is "dry" and can actually be filed immedicatedly. The best part is that the optical properties in the light microscope make the background virtually invisible and the stained structures have greater clarity. Bernice _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From info <@t> instrumedics.com Fri Feb 10 11:13:55 2006 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Feb 10 11:31:47 2006 Subject: [Histonet] mounting media References: <20060210163824.2181.qmail@web50903.mail.yahoo.com> Message-ID: <001601c62e65$66672c50$6401a8c0@INSTRUMEDICS22> Cheryl CureMount is dispensed just like Permount. The cover glass is placed over the slide making sure not to trap bubbles as one usually does. The slide is then placed under a UV lamp for 20 sec. The cover slip will be fixed in place and the slide is ready to be viewed in the microscope We have had no indication that there is any storage problem. CureMount I for frozen sections and CureMount II for paraffin sections comes in a 60ml amber dropping bottle. The starter kit of 2 bottles of CureMount and a UV lamp is reasonably priced . Bernice ----- Original Message ----- From: Cheryl To: Instrumedics ; HistoNet Server Sent: Friday, February 10, 2006 11:38 AM Subject: Re: [Histonet] mounting media I'd like to learn more about this-- Is this used with coverslips or like flo-tex (sp?) is used--poured over a slide? What are the storage statistics (temp/humidity/longevity) and can we try it without breaking the budget or a patented UV source?? Instrumedics wrote: I read the interesting review of mounting media by Tony Collins in Microscopy Today. I regret that CureMount, which is an Instrumedics product was not evaluated. was developed to have a refractive index that matches that of dehydrated and cleared frozen and paraffin sections. It contains a photocatalyst which makes it possible to "cure" the CureMount with 20 sec UV exposure . The slide is "dry" and can actually be filed immedicatedly. The best part is that the optical properties in the light microscope make the background virtually invisible and the stained structures have greater clarity. Bernice _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Fri Feb 10 11:40:50 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Feb 10 11:40:57 2006 Subject: [Histonet] A multi-use simple blocking solution?.... In-Reply-To: <6.0.0.22.1.20060210084254.01b48bb0@gemini.msu.montana.edu> Message-ID: <200602101740.k1AHek3t028072@chip.viawest.net> For blocking in general I use a serum free protein block before the primary antibody and then again before the secondary. If I still experience problems with non specific binding (usually of the secondary, but not always), I will go more aggressive by using serum block matched to the host of the secondary (usually a 10% solution without BSA) and if this still does not do the trick I add to the serum block 10% normal sera from the species I am trying to label. (Human serum is added to the serum block for human tissue samples, mouse serum added to the block for mouse tissue samples, etc.) You must be careful not to block all of the antigen sites on your tissue, though. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Friday, February 10, 2006 8:46 AM To: Glenn Krasinski; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] A multi-use simple blocking solution?.... We prefer to use normal serum blocking, with the serum matched to the host of the secondary and do not use BSA based blockers. This has been discussed at great lengths on Histonet, check out Histonet Archives. Blocking, per your question, is done many ways. I suggest you do to the DAKO website, information and access the Handbook of Immunochemical Methods, Boesnish discusses various blocking agents, and you can download this wonderful freebie in pdf format. You can buy superblocks ready to use from various companies, i.e.At 03:13 PM 2/9/2006, you wrote: >I have been looking through the protocols on some of the IHC databases >for blocking solutions. Some of the recipes are fairly exotic (fish >skin gelatin?). Is there a good simple blocking solution that can be >used in a variety of applications (primary/secondary)? How about 1x PBS with 1% BSA? > > Many thanks. > > Glenn M. Krasinski > San Diego, CA > > >--------------------------------- > > What are the most popular cars? Find out at Yahoo! Autos >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Fri Feb 10 11:40:32 2006 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Feb 10 11:41:16 2006 Subject: [Histonet] Labvision Neomarkers GCDFP-15 Ab1, Clone 23A3 Message-ID: We use Signet's GCDFP-15 mAb at 1:100 for 30' with good results. A 1:25 dilution sounds "expensive"! Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Theresa Rohr" 02/10/06 12:03PM >>> Dear Histonetters, I am using Neomarker's GCDFP Ab-1, Clone 23A3 at a 1:25 titer. I use a 6.0 citrate for Target Retrieval for 20 minutes in a Steamer and 20 minute cooling. I stain on the Dako Autostainer with Envision+ at 30 minutes in Primary Ab, 30 minutes in Polymer and 5 minutes in DAB. The staining results are just OK nothing extraordinary. Has anyone used this antibody and had great results? If so, what are you doing differently than my protocol? Do you think a 9.0 EDTA buffer for Retrieval would be better? What about increasing my time in DAB? Any help would be greatly appreciated. I have emailed their techservice but thought my peers might have more to teach me. Thanks so much, Theresa Rohr rohrt@nyackhospital.org Theresa Rohr, BA HT(ASCP) Section Head, Histology Nyack Hospital 160 North Midland Avenue Nyack, New York 10960 phone - 845-348-2276 fax - 845-348-8430 Nyack Hospital is required by Law to protect the privacy of health information under HIPAA Rules and Regulations Part 11, Article 45 CFR, Parts 160-164. This email, including attachments, is intended for the exclusive use of the person or the entity to which it is addressed. It may contain confidential or legally protected information. The authorized recipient is prohibited from disclosing this information to any other party and is required to destroy this information after its stated need has been fulfilled. If the recipient or reader of this email is not the intended recipient or her/his authorized agent, the reader is hereby notified that any dissemination, distribution or copying of this email is phohibited. If you believe that you have received this email in error, please advise the sender immediately by reply email and then delete this email immediately. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From David.Muskett <@t> RLC.NHS.UK Fri Feb 10 11:57:21 2006 From: David.Muskett <@t> RLC.NHS.UK (Muskett David) Date: Fri Feb 10 11:57:34 2006 Subject: [Histonet] Unsubscribe Message-ID: <5B074C3F334D2F449497BAB0CAEAD65507C46A@AHMAIL01.xalderhey.com> Please remove me from the list server. Thanks David David Muskett Chief Biomedical Scientist, Histology Royal Liverpool Children's NHS Trust Eaton Road Liverpool L12 2AP Internal telephone x3615 Telephone (0151) 293 3656 Fax (0151) 293 3617 http://nhsald02/histopathology From LJohns53 <@t> CNTUS.JNJ.COM Fri Feb 10 12:04:06 2006 From: LJohns53 <@t> CNTUS.JNJ.COM (Johns, Laura [CNTUS]) Date: Fri Feb 10 12:04:22 2006 Subject: [Histonet] background in lymphoid tissue Message-ID: <70F83FE9F65318468A612768E7043F89037540E8@cntusmaexs9.na.jnj.com> Hi All, I am having a problem with background staining in lymphoid tissue. I have tried increasing the time of my peroxide block, and adding in an avidin/biotin blocking step, but still there is some slight staining in my negative controls (no primary or secondary antibodies). I have noticed this in spleen and in other organs where there is an area of inflammation, and with several different antibody combinations. Has anyone experienced this problem? Any suggestions for getting rid of it? Thanks! Laura Laura M. Johns, Ph.D. Centocor, Inc. 145 King of Prussia Road (Mail Stop # R-4-2) Radnor, PA 19087 Phone: 610-240-8284 .... Fax: 610-240-8150.... Email: ljohns53@cntus.jnj.com > Confidentiality Notice: This message is intended for the individual(s) or > entity to which it is addressed and may contain information that is > privileged, confidential and exempt from disclosure under applicable law. > If the reader of this message is not the intended recipient, he/she is > hereby notified that any dissemination, distribution or copying of this > communication is strictly prohibited. If you have received this > communication in error, please notify the sender by telephone and delete > the e-mail from your system immediately. Thank you. > > From LJohns53 <@t> CNTUS.JNJ.COM Fri Feb 10 12:26:30 2006 From: LJohns53 <@t> CNTUS.JNJ.COM (Johns, Laura [CNTUS]) Date: Fri Feb 10 12:26:45 2006 Subject: [Histonet] background in lymphoid tissue Message-ID: <70F83FE9F65318468A612768E7043F89037540EF@cntusmaexs9.na.jnj.com> Hi Liz, Thanks for your response. I am using normal goat serum in place of the primary and secondary antibodies. I dilute my primary in NGS, and my secondary in diluent. I haven't tried adding in another protein block or power block; I really thought the avidin/biotin would take care of it, but it didn't, and I haven't had a chance to try anything else yet. Any suggestions are welcome! Laura -----Original Message----- From: Liz Beitman [mailto:LBeitman@cellmarque.com] Sent: Friday, February 10, 2006 1:18 PM To: Johns, Laura [CNTUS] Subject: RE: [Histonet] background in lymphoid tissue Hi Laura, Just curious here, what might you be using in place of applying antibody to the negative? Might you be using a mouse or rabbit negative serum? Sometimes these serums are more sensitive than if you just used diluent. There are other types of blockers as well such as protein blocker that blocks for any endogenous proteins that can be inherent to certain tissues. Have you tried a different blocker? I hope you don't mind my response and if there is anything I can do to help, please feel free to give me a call anytime. Sincerely, Liz Beitman Cell Marque 1-800-665-7284 x 19 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Johns, Laura [CNTUS] Sent: Friday, February 10, 2006 12:04 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] background in lymphoid tissue Hi All, I am having a problem with background staining in lymphoid tissue. I have tried increasing the time of my peroxide block, and adding in an avidin/biotin blocking step, but still there is some slight staining in my negative controls (no primary or secondary antibodies). I have noticed this in spleen and in other organs where there is an area of inflammation, and with several different antibody combinations. Has anyone experienced this problem? Any suggestions for getting rid of it? Thanks! Laura Laura M. Johns, Ph.D. Centocor, Inc. 145 King of Prussia Road (Mail Stop # R-4-2) Radnor, PA 19087 Phone: 610-240-8284 .... Fax: 610-240-8150.... Email: ljohns53@cntus.jnj.com > Confidentiality Notice: This message is intended for the > individual(s) or entity to which it is addressed and may contain > information that is privileged, confidential and exempt from disclosure under applicable law. > If the reader of this message is not the intended recipient, he/she is > hereby notified that any dissemination, distribution or copying of > this communication is strictly prohibited. If you have received this > communication in error, please notify the sender by telephone and > delete the e-mail from your system immediately. Thank you. > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Fri Feb 10 13:54:44 2006 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Feb 10 13:55:18 2006 Subject: [Histonet] "Let's Go Histo" motto for 2007 NSH Message-ID: Be careful or she will chase you with her Hi-Profiles :>) Robyn From sjchtascp <@t> yahoo.com Fri Feb 10 14:06:51 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Feb 10 14:07:00 2006 Subject: [Histonet] Miles Cryostat repair Message-ID: <20060210200651.32067.qmail@web90208.mail.scd.yahoo.com> I'm looking for someone in the Madison, WI arear that works on these older "Ames" cryostats. Steve --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! From mtitford <@t> aol.com Fri Feb 10 14:37:59 2006 From: mtitford <@t> aol.com (mtitford@aol.com) Date: Fri Feb 10 14:38:21 2006 Subject: [Histonet] Learning histology Message-ID: <8C7FC9904048E67-FC4-9C6@FWM-D44.sysops.aol.com> Stephen at Texas A & M wants to know how to learn histology. I think the best way is to borrow a set of teaching microscope slides from your anatomy department (or cellular biology, whatever they call it now) and sit down with a microscope. 1) First of all study the different types of cells ( three types of muscle cells, the connective tissue cells, blood cells and developing cells, all the epithelial cells, glandular cells, etc) 2) Then study the organ systems (Kidney, lung, intestine, nervous system, G.I. Tract, endocrine etc).Learn to diffentiate the different parts of the stomach, large and small intestines, brain, etc. Nearly every organ has one or two key structures that "give it away". Try to learn those. 3) Then learn the tricks professors play on you: be able to differentiate resting breast from prostate, lactating breast from thyroid, spleen from lymph node, etc. This is a time consuming method but it pays off. Modern histology classes are often taught using powerpoint or some other method where they use the best d*** photographs they can find of that organ or structure. In real life you have to look around the slide, orient yourself, find identfying features in the tissue, and then name it! Career histotechnologists add an additional step of learning the special stains associated with each particular organ or tissue. Good luck with your studies Mike Titford USA Pathology Mobile AL USA From pruegg <@t> ihctech.net Fri Feb 10 15:03:08 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Feb 10 15:03:16 2006 Subject: [Histonet] background in lymphoid tissue In-Reply-To: <70F83FE9F65318468A612768E7043F89037540E8@cntusmaexs9.na.jnj.com> Message-ID: <200602102103.k1AL343t016411@chip.viawest.net> Laura, It can be very difficult to quench endogenous peroxidase in some tissue/cells, neutrophils in spleen and inflamation are notorious for being difficult to quench. If you can you might try using an alk.phos detection system to see if it is the endogenous peroxidase giving you these problems. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Johns, Laura [CNTUS] Sent: Friday, February 10, 2006 11:04 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] background in lymphoid tissue Hi All, I am having a problem with background staining in lymphoid tissue. I have tried increasing the time of my peroxide block, and adding in an avidin/biotin blocking step, but still there is some slight staining in my negative controls (no primary or secondary antibodies). I have noticed this in spleen and in other organs where there is an area of inflammation, and with several different antibody combinations. Has anyone experienced this problem? Any suggestions for getting rid of it? Thanks! Laura Laura M. Johns, Ph.D. Centocor, Inc. 145 King of Prussia Road (Mail Stop # R-4-2) Radnor, PA 19087 Phone: 610-240-8284 .... Fax: 610-240-8150.... Email: ljohns53@cntus.jnj.com > Confidentiality Notice: This message is intended for the > individual(s) or entity to which it is addressed and may contain > information that is privileged, confidential and exempt from disclosure under applicable law. > If the reader of this message is not the intended recipient, he/she is > hereby notified that any dissemination, distribution or copying of > this communication is strictly prohibited. If you have received this > communication in error, please notify the sender by telephone and > delete the e-mail from your system immediately. Thank you. > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Fri Feb 10 15:10:58 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Feb 10 15:11:14 2006 Subject: [Histonet] Mer2 antibody Message-ID: <200602102110.k1ALAs3t018203@chip.viawest.net> Has anyone tried Mer (N-19) sc6872 on ffpe tissues? This is UFO or ARK family of receptor tyrosine kinases also designated as Mer, Nyk or Eyk. Mer may be involved in the development of glioblastomas so we are using it on human brain tissue ffpe. I asked this question of the IHC Resource Group www.ihcrg.org but got no responses, so I thought I would put it out to histonet. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From lharris <@t> samhealth.org Fri Feb 10 16:01:31 2006 From: lharris <@t> samhealth.org (lharris@samhealth.org) Date: Fri Feb 10 16:01:57 2006 Subject: [Histonet] RE: purple blue haze Message-ID: <5B3B26C4B71D5E469892D1860ABE10EA7F1E8D@SHSEXVS01.int.samhealth.net> We had a purple blue haze on our H&E's when we used cold tap water run through the stainer. We had to use hot water and the haze went away. I've tried to return to using cold water and we get the haze every time. Lori A. Harris, HT (ASCP) Histology Section Leader GSRMC - Pathology 3600 NW Samaritan Drive Corvallis, OR 97330 1-541-768-6078 lharris@samhealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, December 21, 2005 8:06 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 25, 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-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@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. online journals (Malcolm McCallum) 2. Purple blue haze (Heckford, Karen - SMMC-SF) 3. Re: online journals (John A. Kiernan) 4. Re:Sections coming off! (Shannon Bryce) 5. re: MMA sections coming off! (Tracey Couse) 6. Re 17. fluorescence immunohistochemistry (Donna Harclerode) 7. Re: Purple blue haze (Andrea Grantham) 8. RE: Purple blue haze (Laurie Colbert) 9. Re: anti-human VEGF antibody (pruegg@ihctech.net) 10. Re: 17. fluorescence immunohistochemistry (John Kiernan) 11. Re: non-specific staining IHC (Malam Jacqueline) 12. MMA sections falling off (Nancy W. Troiano) 13. Modified Davidson's Rat Eyes and Testes Fixation Times (Sharon E Willman) 14. Embedding Centers (Roberta Horner) 15. Glass coverslippers (Angela Bitting) 16. RE: Glass coverslippers (Bartlett, Jeanine) 17. Re: Embedding Centers (Rene J Buesa) 18. San Antonio Histology connection (Jackie M O'Connor) 19. RE: Embedding Centers (Anne Van Binsbergen) 20. RE: Glass coverslippers (Anne Van Binsbergen) 21. Re: MMA sections falling off (Gayle Callis) 22. Re: Embedding Centers (Pamela Marcum) 23. decal rapid (Sharon Allen) 24. FW: artifact (Santana, Diane) 25. antibody search (Steven Coakley) 26. Mouse liver tissue processing (Luis Chiriboga) 27. RE: Mouse liver tissue processing (HSRL) ---------------------------------------------------------------------- Message: 1 Date: Tue, 20 Dec 2005 12:11:01 -0600 From: "Malcolm McCallum" Subject: [Histonet] online journals To: , , , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hi Which of the below three options do you think is best regarding a journal with both print and online versions???? 1) Journal is open access, the publisher will charge page charges to authors. 2) No page charges (except color plates), but the publisher charges download fees and only the abstracts are open access. 3) Page charges are optional for the author, if the author pays the entire article is open access, if not then only the abstract will be accessible without paying a download fee. In case someone is unaware, If an article is entirely open access then anyone can read it or download it online. IF only the abstract is available then you can read the abstract online, but must pay a fee to download the entire article. Thanks for the feedback, its actually very important! Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html ------------------------------ Message: 2 Date: Tue, 20 Dec 2005 11:17:21 -0700 From: "Heckford, Karen - SMMC-SF" Subject: [Histonet] Purple blue haze To: "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Dear Histonetters, Has anyone had any problems with a purple- blue haze on their H&E using Richard Allen Scientific Hematoxylin 7211. I am currently filtering it. I am still getting the haze. I have changed nothing, it just appeared a couple of weeks ago. Any help would be greatly appreciated. Happy Holidays, Karen Heckford HT (ASCP) CE Lead Histology Technician St. Mary's Medical Center Histology Department 450 Stanyan St. San Francisco, Ca. 94117 1-415-668-1000 ext. 6167 email: kheckfor@chw.edu ------------------------------ Message: 3 Date: Tue, 20 Dec 2005 13:42:34 -0500 From: "John A. Kiernan" Subject: Re: [Histonet] online journals To: Malcolm McCallum Cc: histonet@lists.utsouthwestern.edu, tws-l@list.wildlife.org, ecolog-l@listserv.umd.edu, parc@listserv.uga.edu Message-ID: <43A8509A.9B855C2A@uwo.ca> Content-Type: text/plain; charset=us-ascii You raise some interesting points. Clearly you're polling a wide range of scientists; please send out a summary when you've analysed the results. Here's my two-centsworth. Paying to read the full text doesn't apply if your institution subscribes to the journal (or more usually to a large batch from the publisher). Since most people who want the full text are in institutions with libraries (or with employers such as drug companies that will pay for individual articles), avoiding page charges is the way to go. Why should the researcher and author pay the publisher? It should be the other way round. Prestigious journals commonly do charge authors, but anyone publishing in such a journal is sure to have a big enough research grant to cover the fees, which are small compared with salaries and items for the lab. Some journals published by learned societies do not charge authors anything, even for coloured pictures if these are needed to make a point. Three examples that come to mind are the Journal of Anatomy, the Journal of Histochemistry and Cytochemistry, and Biotechnic & Histochemistry. I'm sure there are many others. All three that I mentioned come free with membership in the society. The cost works out at about $5 per issue (print, and access online). I've not heard of the option of the author paying and the reader not paying. It's hard to see that working; even non-profit societies cannot run their journals at a loss. Libraries have to pay much more than individual members. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Malcolm McCallum wrote: > > Hi > Which of the below three options do you think is best regarding a journal with both print and online versions???? > > 1) Journal is open access, the publisher will charge page charges to authors. > 2) No page charges (except color plates), but the publisher charges download fees and only the abstracts are open access. > 3) Page charges are optional for the author, if the author pays the entire article is open access, if not then only the abstract will be accessible without paying a download fee. > > In case someone is unaware, If an article is entirely open access then anyone can read it or download it online. IF only the abstract is available then you can read the abstract online, but must pay a fee to download the entire article. > > Thanks for the feedback, its actually very important! > > Malcolm L. McCallum > Assistant Professor > Department of Biological Sciences > Texas A&M University Texarkana > 2600 Robison Rd. > Texarkana, TX 75501 > O: 1-903-233-3134 > H: 1-903-791-3843 > Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Tue, 20 Dec 2005 13:01:17 -0600 From: "Shannon Bryce" Subject: [Histonet] Re:Sections coming off! To: "Joanne Mauger" , , Message-ID: <43A8009D0200009900000BC0@swnw124.swmed.edu> Content-Type: text/plain; charset=US-ASCII We get our slides which are the + coated slides from Fisher. Our MMA sections are also falling off here lately so I went to another department and borrowed a box of theirs that they bought through VWR and have had no problems so I would guess it's a bad lot.....anyone have lot numbers? Thanks, Shannon Bryce UT Southwestern Medical Center >>> "Joanne Mauger" 12/20/05 8:39 AM >>> Hi Patricia, Are you using slides from Erie? We buy ours through Fisher, and have been having trouble keeping sections on + slides as well. It is as if they have no charge at all. We think the lot may be bad. The company claims to have no complaints. Anyone else having similar problem? Jo Mauger >>> "Patricia F Lott" 12/19/05 3:21 PM >>> Help! We are losing our thin plastic sections! We have cut PMMA plastic sections at 5 microns and put them on "+" slides, and the sections fall off either during de-plasticizing or during staining! We have tried: Haupt's adhesive, longer drying times, "+" slides, plus slides and Haupt's together, etc....no luck! Any suggestions would be welcome! _______________________________________________ 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 ------------------------------ Message: 5 Date: Tue, 20 Dec 2005 15:15:42 -0500 From: Tracey Couse Subject: re: [Histonet] MMA sections coming off! To: histonet@lists.utsouthwestern.edu Message-ID: <43A8666E.2030005@ibb.gatech.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Patricia, Try manually coating plain glass slides with poly-l-lysine, chrome/gelatin, or glycerin/gelatin. We use all of these for our thin plastic sections. I also suggest setioning a micron or two thinner (cut at 3 or 4um). A thinner section will reduce the surface area to volume ratio and may promote better section adherance. You may need to dry the sldies using a slide press and dry longer or with higher temperatures. Also, try searching the Histonet archives for alternative slide coating suggestions. Good luck! -- Tracey Couse Laboratory Coordinator Georgia Tech/Emory Center for the Engineering of Living Tissues Georgia Institute of Technology IBB, Room 1123 Office: 404.385.2611 Lab: 404-385-6735 Fax: 404.894.229 Date: Mon, 19 Dec 2005 14:21:07 -0600 From: "Patricia F Lott" Subject: [Histonet] MMA sections coming off! To: Message-ID: Content-Type: text/plain; charset="us-ascii" Help! We are losing our thin plastic sections! We have cut PMMA plastic sections at 5 microns and put them on "+" slides, and the sections fall off either during de-plasticizing or during staining! We have tried: Haupt's adhesive, longer drying times, "+" slides, plus slides and Haupt's together, etc....no luck! Any suggestions would be welcome! 1 ------------------------------ Message: 6 Date: Tue, 20 Dec 2005 13:06:17 -0800 From: "Donna Harclerode" Subject: [Histonet] Re 17. fluorescence immunohistochemistry To: , Cc: Adam Johnson Message-ID: <3DE0F644E093DF4BAE80C254176696A505B869@mp-mailserver.macropore.com> Content-Type: text/plain; charset="US-ASCII" Message: 17 Date: Mon, 19 Dec 2005 22:08:48 +0100 From: Emily.Wiesner@medecine.unige.ch Subject: [Histonet] fluorescence immunohistochemistry To: histonet@lists.utsouthwestern.edu Message-ID: <1135026528.43a72160c086f@webmail.medecine.unige.ch> Content-Type: text/plain; charset=ISO-8859-1 Hi All, I am new to fluorescence immunohistochemistry and I was wondering if anyone can let me know the simple steps involved in performing this technique. I know the dilutions required for the primary and secondary antibodies, I just need to known what solutions are used for washes and the steps in between! Thanks in advance, Emily Dear Emily I do very simple IHC using fluorescent secondaries from Jackson Immunoresearch and purified primary abs. Depending on what you are doing, there can be more steps added. I usually am staining sections on a slide or cells on chamber slides or in a plate. For the unfixed frozen sections on slides I normally would fix 5 min in 75% acetone 25% ethanol and then right into PBS- do not allow the fixed slides to dry. The only change would be if the antibody prefers formalin fixation to acetone/ alcohol (thanks Gayle- I love this fixative for almost everything). I load them into a Sequenza(tm) rack (Thermo Electron, formerly Shandon) or you could put them in a humid chamber of some sort, wiping excess buffer before adding antibody or blocking solutions. OPTIONAL I have recently started using Image-iT(tm) FX signal enhancer, I36933, $80 Invitrogen for auto fluorescence (not sure it helps yet, but sure does not hurt) 30 minutes. I would not use this on routinely- my sections have infracted areas that have been a problem in the past) After blocking, I rinse 1 time in the Sequenza rack (3 x 5 minutes if you are doing them by hand) Add 100ul of the antibody diluted in Dako antibody diluent S0809- usually 1:100- 1:1k (dilutions are antibody dependant, but I have found in fluorescence too high a concentration is not as big a problem as with HRP formats) If you are using a humid chamber be sure to cover all the tissue. I have incubated slides at Room temperature for 1-2 hours, but now am staining overnight in the fridge (not so good in clinical situations) After primary incubation, I rinse 1 time in the Sequenza rack (3 x 5 minutes if you are doing them by hand) Next is Jackson secondary at a 1:100 dilution (I use Cy3 conjugates for single color and FITC and RITC for dual color. All my secondaries are minimal cross (mouse to rat), but if all your tissue is human that is not necessary. 30 minutes I add DAPI to my secondaries to give a blue nuclear stain so I can tell what I am looking at and localize the stain. Rinse again 3 times in PBS and coverslip with your choice of aqueous media. For media that never gets hard, I prefer Immunomount (Thermo Electron, formerly Shandon) or Aqua Mount (Lerner from VWR) same thing different label or the Prolonge Mounting media P36930 for $101 that will harden. I do not like the hard mount media from Vector- I got many air bubbles forming day after the sldes were coverslipped. For cells on plates or chamber slides I usually fix with 10% NBF or 4% PFA (same concentration of fixative, 4%PFA will contain no methanol) for 5-30 minutes. CD makers can be very hard to stain with NBF but with a short fix, I have been pretty luck so far. I rinse 3 x 10 minutes with PBS I have not used the blocking reagents on cells as I have not seen any autofluorescence in our cardiac myocytes, neurons or stem cells. The rest of the procedure is the same except in a 6 well plate you need a lot of antibody to cover the plate (about 600u). I try to use a rocker if it is available to help to be sure of good coverage. I do not usually cover my slides or plates, but if your room is very bright or your signal very week, it would be a good idea to protect the secondaries from light. I like to use isotype controls (negative controls) or rabbit IgGs for much of my work, but in a pinch I use just the antibody diluent to be sure the stain I see, is a result of the primary antibody and not the secondary binding to some part of the cell. In cell assays I always want to stain a known positive and a known negative cell if at all possible in addition to the isotype controls. If you are making your own diluent (to me Dako diluent is great and I have used it successfully for over 10 years in many applications) I use 0.3% Triton, 5 % Normal serum from the species that your secondary is made in - (all my secondaries are made in donkey so I would use donkey serum) in PBS. I have used donkey secondaries when possible because when I tested them years ago, they were a bit cleaner on the CD markers I was using. Dako diluent must have a similar make up, but it lasts a year in the fridge and it permeablizes intact cells very nicely. One big thing when purchasing fluorophores, make sure the filters on your scope can see the secondaries you are purchasing. I found out at my current job after staining with Cy3 and RITC that the reason I could not get any stain was they had a Texas red filter not the Cy3, RITC filter. Ideally if you are doing multicolor you want narrow band filters so you can separate the colors, with broad pass filters the colors blur together and it is hard to separate the results. Good Luck! Donna Donna Harclerode, HT, (ASCP), HTL, QIHC Scientist / Immunohistochemistry Cytori Therapeutics 3020 Callan Rd. San Diego, CA 92121 858-458-0900 ext 322 dharclerode@cytoritx.com ------------------------------ Message: 7 Date: Tue, 20 Dec 2005 14:19:47 -0700 From: Andrea Grantham Subject: Re: [Histonet] Purple blue haze To: "Histonet (E-mail)" Message-ID: <4.3.2.7.2.20051220140523.00ce4520@algranth.inbox.email.arizona.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Karen, Yes I have recently noticed the "haze" of the H&E stained slides. I have been using this hematoxylin for years and this is the first time that I have had a problem with it. Like you I have filtered it but it hasn't seemed to help. I also have been using their Type 6 paraffin and in the last few months have noticed that it blows apart on the waterbath. I have lowered the temp. but it didn't help -> 35-39 degrees C. A different lot number that they sent to me was only marginally better. A couple of weeks ago one of my clients brought a project consisting of blocks that had been processed and embedded here over 4-5 years. The paraffin throughout was RAS Type 6. The blocks from the first 4 years cut fine and laid out on the waterbath fine but when I got to recently embedded blocks they started the rapid expanding and blowing apart when they were placed on the waterbath. A histotech in CA told me they have been having the same problem in their lab. Hmmmmm.....could they have changed the formula? Maybe this is so with the Hematoxylin as well. I'm trying different paraffins looking for one comparable to the Type 6 only more stable on the waterbath. I really don't want to do this with Hematoxylin because my clients love my staining but I hate ugly slides. Andi At 11:17 AM 12/20/2005 -0700, Heckford, Karen - SMMC-SF wrote: >Dear Histonetters, >Has anyone had any problems with a purple- blue haze on their H&E using >Richard Allen Scientific Hematoxylin 7211. I am currently filtering it. I >am still getting the haze. I have changed nothing, it just appeared a >couple of weeks ago. Any help would be greatly appreciated. > >Happy Holidays, > > >Karen Heckford HT (ASCP) CE >Lead Histology Technician >St. Mary's Medical Center >Histology Department >450 Stanyan St. >San Francisco, Ca. 94117 >1-415-668-1000 ext. 6167 >email: kheckfor@chw.edu > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html ------------------------------ Message: 8 Date: Tue, 20 Dec 2005 15:12:17 -0800 From: "Laurie Colbert" Subject: RE: [Histonet] Purple blue haze To: "Andrea Grantham" , "Histonet (E-mail)" Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628006307C76@EXCHANGE1.huntingtonhospital.com> Content-Type: text/plain; charset="iso-8859-1" Your discussion rang a bell with me, and I've been meaning to call Richard Allan. We use their Type 9 paraffin, and for the last 3 or 4 months it also has been "blowing apart on the waterbath." We lowered the temp on the waterbaths, and this seems to have helped. Our housekeeping personnel have even noticed a difference in the paraffin (scraping it off the floor). They say it is stickier lately. I think something has been changed. Laurie Colbert Huntington Hospital Pasadena, CA -----Original Message----- From: Andrea Grantham [mailto:algranth@u.arizona.edu] Sent: Tuesday, December 20, 2005 1:20 PM To: Histonet (E-mail) Subject: Re: [Histonet] Purple blue haze Karen, Yes I have recently noticed the "haze" of the H&E stained slides. I have been using this hematoxylin for years and this is the first time that I have had a problem with it. Like you I have filtered it but it hasn't seemed to help. I also have been using their Type 6 paraffin and in the last few months have noticed that it blows apart on the waterbath. I have lowered the temp. but it didn't help -> 35-39 degrees C. A different lot number that they sent to me was only marginally better. A couple of weeks ago one of my clients brought a project consisting of blocks that had been processed and embedded here over 4-5 years. The paraffin throughout was RAS Type 6. The blocks from the first 4 years cut fine and laid out on the waterbath fine but when I got to recently embedded blocks they started the rapid expanding and blowing apart when they were placed on the waterbath. A histotech in CA told me they have been having the same problem in their lab. Hmmmmm.....could they have changed the formula? Maybe this is so with the Hematoxylin as well. I'm trying different paraffins looking for one comparable to the Type 6 only more stable on the waterbath. I really don't want to do this with Hematoxylin because my clients love my staining but I hate ugly slides. Andi At 11:17 AM 12/20/2005 -0700, Heckford, Karen - SMMC-SF wrote: >Dear Histonetters, >Has anyone had any problems with a purple- blue haze on their H&E using >Richard Allen Scientific Hematoxylin 7211. I am currently filtering it. I >am still getting the haze. I have changed nothing, it just appeared a >couple of weeks ago. Any help would be greatly appreciated. > >Happy Holidays, > > >Karen Heckford HT (ASCP) CE >Lead Histology Technician >St. Mary's Medical Center >Histology Department >450 Stanyan St. >San Francisco, Ca. 94117 >1-415-668-1000 ext. 6167 >email: kheckfor@chw.edu > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Tue, 20 Dec 2005 17:56:17 -0800 (PST) From: pruegg@ihctech.net Subject: Re: [Histonet] anti-human VEGF antibody To: "Katri Tuomala" Cc: Histonet Message-ID: <43434.207.200.116.10.1135130177.squirrel@207.200.116.10> Content-Type: text/plain;charset=iso-8859-1 I am using mouse anti-vegf from Santa Cruz with good results on human and baboon ffpe tissues, it is not easy, but I do HIER in a water bath in high ph buffer at 85dc for 3 hrs., followed by overnight incubation of the primary 1:200, detected with mouse labeled polymer (I use Envision from DAKO). Patsy > Andrea, > I cannot say this is the best anti-VEGF antibody, but I am doing a > research > project on FFPE human brain tissue and using placenta as a control with > following antibody: > monoclonal mouse anti human VEGF, clone VG1 from ID Labs (www.idlabs.com), > dilution 1:100, HIER in citrate buffer pH 6.0. Detection system is from > Zymed, Histostain Plus Kit.. > Glioblastoma stains strongly with this antibody. I have not used this with > frozen sections. > > Katri > > Katri Tuomala > Hamilton, Ontario, Canada > ----- Original Message ----- > From: "Andrea T. Hooper" > To: "Histonet" > Sent: Monday, December 19, 2005 8:37 PM > Subject: [Histonet] anti-human VEGF antibody > > >> What's the current consensus on the best antibody to use for anti-human >> VEGF immunostaining on human FFPE sections (frozens as well if >> possible). >> >> Thanks so much!! >> Andrea >> -- >> >> _______________________________________________ >> 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 > ------------------------------ Message: 10 Date: Wed, 21 Dec 2005 02:10:48 -0500 From: John Kiernan Subject: Re: [Histonet] 17. fluorescence immunohistochemistry To: Donna Harclerode Cc: histonet@lists.utsouthwestern.edu, Adam Johnson , Emily.Wiesner@medecine.unige.ch Message-ID: <43A8FFF8.D988E7C6@uwo.ca> Content-Type: text/plain; charset=us-ascii When I was a student in Birmingham, England (that's the Birmingham with the silent h), the immunology and serology lecturer told us: "water is unphysiological and beer is expensive, so we use saline." He must have been a good lecturer because I can still recall that apophthegm 40 years on when afar and asunder! John Kiernan London, Ontario. ------------------------------- > Hi All, > I am new to fluorescence immunohistochemistry and I was wondering if > anyone can let me know the simple steps involved in performing this > technique. I know the dilutions required for the primary and secondary > antibodies, I just need to known what solutions are used for washes and > the steps in between! > Thanks in advance, ------------------------------ Message: 11 Date: Wed, 21 Dec 2005 12:00:58 -0000 From: Malam Jacqueline Subject: [Histonet] Re: non-specific staining IHC To: "Histonet submissions (histonet@lists.utsouthwestern.edu)" Message-ID: Content-Type: text/plain We use the Benchmark XT but make our own prep kits up for neg controls (poly and mono) and are not experiencing any more non-sp staining than we normally do which is minimal. Jacqui DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. ------------------------------ Message: 12 Date: Wed, 21 Dec 2005 07:20:53 -0500 From: "Nancy W. Troiano" Subject: [Histonet] MMA sections falling off To: histonet@lists.utsouthwestern.edu Message-ID: <5.2.1.1.2.20051221071248.00c2c038@email.med.yale.edu> Content-Type: text/plain; format=flowed; charset=us-ascii We use Chrome-alum gel coated slides and prepare them as follows: dissolve 9 g gelatin(Fisher #G8-500) per 1000 ml distilled water (solution A) while gently heating solution. Dissolve 4 g Chromium Potassium Sulfate (Fisher #C337-500) in distilled water (solution B). Prepare gelatin solution for slide dip: Mix 500 ml Solution a with 19.25 ml solution B. Heat to 70-75 Deg. C, dip slides 2 -4 minutes in this solution. Allow to dry vertically, then put in hot oven overnight (60 deg. C). To adhere sections to slides we place the 5 micron MMA sections on slide, spread with 70% ethanol, cover with a piece of plastic and stack slides then clamp with an office clamp. Put slides in 37deg C oven for two nights or for one night in 60 deg C oven (don't do the hot oven if you are subsequently staining for enzymes). For immuno we mount our 5 micron plastic sections on hybridization slides from Scientific Device Laboratory, Cat. #063 and place in hot oven 60 deg C overnite. Hope this helps! ------------------------------ Message: 13 Date: Wed, 21 Dec 2005 07:18:32 -0600 From: Sharon E Willman Subject: [Histonet] Modified Davidson's Rat Eyes and Testes Fixation Times To: histonet@lists.utsouthwestern.edu Message-ID: <43A95628.7060609@bms.com> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Hi, I was needing input on how long you fix rat eyes and testes in Modified Davidson's. Any suggestions would be most appreciated. Thanks in advance, Sharon Willman ------------------------------ Message: 14 Date: Wed, 21 Dec 2005 08:25:26 -0500 From: "Roberta Horner" Subject: [Histonet] Embedding Centers To: Message-ID: Content-Type: text/plain; charset="US-ASCII" My embedding center just died. Last time our maintenance looked at it they couldn't get the parts (its about 17 years old). I need to get information quick on which ones you like and are good. So far I have quotes from Surgipath and on the Leica but I don't know anything about them. Any comments please? Roberta ------------------------------ Message: 15 Date: Wed, 21 Dec 2005 08:49:01 -0500 From: "Angela Bitting" Subject: [Histonet] Glass coverslippers To: Message-ID: Content-Type: text/plain; charset=US-ASCII Can anyone recommend a automated glass coverslipper? I've used Hacker in the past. Any that are faster, better? Thanks. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. ------------------------------ Message: 16 Date: Wed, 21 Dec 2005 08:56:05 -0500 From: "Bartlett, Jeanine" Subject: RE: [Histonet] Glass coverslippers To: "Angela Bitting" , Message-ID: Content-Type: text/plain; charset="us-ascii" We recently demo'd the new one from Leica and it was great! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Angela Bitting Sent: Wednesday, December 21, 2005 8:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Glass coverslippers Can anyone recommend a automated glass coverslipper? I've used Hacker in the past. Any that are faster, better? Thanks. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 17 Date: Wed, 21 Dec 2005 06:12:16 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Embedding Centers To: Roberta Horner , histonet@lists.utsouthwestern.edu Message-ID: <20051221141216.20482.qmail@web61215.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi Roberta: I would suggest you to call Sakura-Finetek. They have a new embedding center that has all the advantages of the old Tissue-Tek, along with the Sakura reliability. Ask for a "demo" instrument (they usually accomodate that type of request). Ren? J. Roberta Horner wrote: My embedding center just died. Last time our maintenance looked at it they couldn't get the parts (its about 17 years old). I need to get information quick on which ones you like and are good. So far I have quotes from Surgipath and on the Leica but I don't know anything about them. Any comments please? Roberta _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 18 Date: Wed, 21 Dec 2005 08:25:29 -0600 From: "Jackie M O'Connor" Subject: [Histonet] San Antonio Histology connection To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Could someone in the San Antonio area contact me directly? Thanks. Jackie O' ------------------------------ Message: 19 Date: Wed, 21 Dec 2005 18:20:37 +0400 From: "Anne Van Binsbergen" Subject: RE: [Histonet] Embedding Centers To: "Roberta Horner" , Message-ID: <7E070A7B959A9F42BE732545E5CF621094ACBA@SKMCEMAIL.skmc.gov.ae> Content-Type: text/plain; charset="us-ascii" Im a fan of the Sakura embedding system - it's ergonomically designed and VERY user friendly Can be 'left handed' or 'right handed' Cassettes left/right Moulds left/right Cold plate left/right Forceps left/right Nothing more annoying (being a lefty) to be forced to work like a non-lefty!! Everything works independently of everything else - ie - micro adjustments can be made to temp in almost any area to your requirements Nice broad anti burn protection to rest hands/fingers while embedding And if you have a VIP5 the basket fits snugly into the holding area - left or right as you prefer of course!! Like I said - I am THE fan of Sakura And I just KNOW that some of you are smiling out there - you know who you are!!! Annieinarabia -----Original Message----- From: Roberta Horner [mailto:rjr6@psu.edu] Sent: Wednesday, December 21, 2005 5:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Embedding Centers My embedding center just died. Last time our maintenance looked at it they couldn't get the parts (its about 17 years old). I need to get information quick on which ones you like and are good. So far I have quotes from Surgipath and on the Leica but I don't know anything about them. Any comments please? Roberta _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 20 Date: Wed, 21 Dec 2005 18:24:12 +0400 From: "Anne Van Binsbergen" Subject: RE: [Histonet] Glass coverslippers To: "Bartlett, Jeanine" , "Angela Bitting" , Message-ID: <7E070A7B959A9F42BE732545E5CF6210948C5F@SKMCEMAIL.skmc.gov.ae> Content-Type: text/plain; charset="us-ascii" Try the Sakura Fast, efficient, small 'footprint' - will fit into a small space Had a Leica once - was so very patient - 2 exchanges (new machines) later - still hated it Annieinarabia -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: Wednesday, December 21, 2005 5:56 PM To: Angela Bitting; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Glass coverslippers We recently demo'd the new one from Leica and it was great! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Angela Bitting Sent: Wednesday, December 21, 2005 8:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Glass coverslippers Can anyone recommend a automated glass coverslipper? I've used Hacker in the past. Any that are faster, better? Thanks. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ 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 ------------------------------ Message: 21 Date: Wed, 21 Dec 2005 08:14:11 -0700 From: Gayle Callis Subject: [Histonet] Re: MMA sections falling off To: "Nancy W. Troiano" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20051221081017.01b39c08@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed To increase the holding power, the gelatin can be 275 bloom, a larger gelatin molecule made from pig collagen. This subbing solution suggested by Nancy is tried and true, also works for the nasty decalcified bone sections, generally larger ones. However, if the subbing solution causes too much blue background staining from hematoxylin, one can dip the subbed slides in NBF a couple of times, rinse well, dry and store in cool, dry place. The NBF crosslinks the gelatin a bit and reduces blue background. At 05:20 AM 12/21/2005, you wrote: >We use Chrome-alum gel coated slides and prepare them as >follows: dissolve 9 g gelatin(Fisher #G8-500) per 1000 ml distilled >water (solution A) while gently heating solution. Dissolve 4 g Chromium >Potassium Sulfate (Fisher #C337-500) in distilled water (solution >B). Prepare gelatin solution for slide dip: Mix 500 ml Solution a with >19.25 ml solution B. Heat to 70-75 Deg. C, dip slides 2 -4 minutes in >this solution. Allow to dry vertically, then put in hot oven overnight >(60 deg. C). To adhere sections to slides we place the 5 micron MMA >sections on slide, spread with 70% ethanol, cover with a piece of plastic >and stack slides then clamp with an office clamp. Put slides in 37deg C >oven for two nights or for one night in 60 deg C oven (don't do the hot >oven if you are subsequently staining for enzymes). For immuno we mount >our 5 micron plastic sections on hybridization slides from Scientific >Device Laboratory, Cat. #063 and place in hot oven 60 deg C >overnite. Hope this helps! > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 22 Date: Wed, 21 Dec 2005 10:14:58 -0500 From: Pamela Marcum Subject: Re: [Histonet] Embedding Centers To: "Roberta Horner" , Message-ID: <6.1.1.1.2.20051221101125.01997318@mail.vet.upenn.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed At 08:25 AM 12/21/2005, Roberta Horner wrote: >My embedding center just died. Last time our maintenance looked at it >they couldn't get the parts (its about 17 years old). I need to get >information quick on which ones you like and are good. So far I have >quotes from Surgipath and on the Leica but I don't know anything about >them. Any comments please? >Roberta >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Hi, I will be totally different as i am buying the Thermo Shandon Embedding Center. I had it in for demo and it had all the features the others have for timing and pre-sets as well as some independent temperature ranges I likes. Also the cold plate is seperate so I can have it on either side or in another area if I want for special projects. Good Luck, I think they all work well it just depends for me on features and price and Shandon also had the right price. Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu ------------------------------ Message: 23 Date: Wed, 21 Dec 2005 09:17:14 -0600 From: Sharon Allen Subject: [Histonet] decal rapid To: "Histonet (E-mail)" Message-ID: <36FE435D6D2F1D489174B22362A961B66B830B@hscxntmx0006.hsc.mb.ca> Content-Type: text/plain; charset="iso-8859-1" Hi, Does anyone know about or what supplier "Decal Rapid" by "Pathtech" can be purchased from? We had used it a few years ago but have no idea where we got it from. I tried searching the internet & came up with Pathtech in Australia but would like a supplier a little closer. Thanks in advance Sharon sallen@hsc.mb.ca This e-mail and/or any documents in this transmission is intended for the address(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. ------------------------------ Message: 24 Date: Wed, 21 Dec 2005 10:27:06 -0500 From: "Santana, Diane" Subject: [Histonet] FW: artifact To: "'Histonet@lists.utsouthwestern.edu'" Message-ID: <4C96AA62BADFD81180CB0002A5AD67FB04D7E282@MAILPMA> Content-Type: text/plain; charset="iso-8859-1" > -----Original Message----- > From: Santana, Diane > Sent: Tuesday, December 20, 2005 10:20 AM > To: 'histonet@lists.utsouthwestern.edu' > Subject: artifact > > I am having a problem with artifact on some of my GI bx's. Actually a > paper published by Anatech "The Innovator", summer2004 shows this same > artifact on the front page. They believe it is a fixation problem, and or > retrieval by the endo Drs. My pathologist rules both these ideas out. I > tend to agree with him, at lease on the fixation. Our bx's go into the > processor around 5 pm and the bx run starts around 2 am. > It is a loss of nuclear detail. This artifact can be on a small section of > a bx. Even if there is 5 bx's in one cassette maybe 1 will have this > artifact where the others look great. I thought at first it was a clearing > problem or nuclear bubbling I changed times in clear rite and oven. It did > not eliminate the problem. My pathologist has seen this by other labs > also, but very seldom. For us it could be 1 or 2 slides a day. Sometimes > not at all. Weekend or weekdays. No consistently. The best way I can > describe it is that it looks like a smudge. > Our bx run is this; > > 10% NBF 30 min ea x2 > 80% Alcohol 10mins > 95% alcohol 10 mins ea x2 > 100% alcohol 17 mins ea x3 > Clear rite 22 mins ea x2 > Paraffin 15 mins ea x3 > > I would appreciate any feedback on this. > Diane Santana > Pentucket Medical Assoc. > Haverhill, Mass. ------------------------------ Message: 25 Date: Wed, 21 Dec 2005 07:22:41 -0800 (PST) From: Steven Coakley Subject: [Histonet] antibody search To: Histonet@lists.utsouthwestern.edu Message-ID: <20051221152241.11885.qmail@web90204.mail.scd.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Gooday everyone, I'd like to get a few recommendations for anti-insulin and anti-glucagon antibodies for FFPE mouse tissue. If anyone has used a specific companies antibodies that work well please let me know. I'm still learning this research IHC stuff so I can use all the help I can "muster" Steve __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 26 Date: Wed, 21 Dec 2005 09:54:28 -0500 From: Luis Chiriboga Subject: [Histonet] Mouse liver tissue processing To: Histonet Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hi everyone We are having a real serious problem getting good sections from mouse livers. The tissue curls and shreds out of the block as soon as it comes in contact with the knife, end up with a section that has the exact outline of the tissue but no tissue. The tissue consistency could best be described as "dry?" Interestingly, it is only the normal livers that have this problem. We have tried sectioning thick, thin, no soak, cold soak......but out of ideas. Anyone have any suggestions? Unfortunately we have no control over the tissue processing, but perhaps can convince to change. Thanks Luis Happy Holidays to all!! ------------------------------ Message: 27 Date: Wed, 21 Dec 2005 11:02:33 -0500 From: "HSRL" Subject: RE: [Histonet] Mouse liver tissue processing To: "'Luis Chiriboga'" , Message-ID: <000c01c60647$f47a7ab0$0200a8c0@HSRLMAIN> Content-Type: text/plain; charset="us-ascii" Luis, Have a tried keeping them on a WET ice for 45 minutes to an hour? If you have tried it and it doesn't work, it is a processing problem that you must rectify. -Tom Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 5930 Main Street Mount Jackson, Virginia 22842 540.477.4440 540.477.4448 tomgalati@hsrl.org www.hsrl.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Luis Chiriboga Sent: Wednesday, December 21, 2005 9:54 AM To: Histonet Subject: [Histonet] Mouse liver tissue processing Hi everyone We are having a real serious problem getting good sections from mouse livers. The tissue curls and shreds out of the block as soon as it comes in contact with the knife, end up with a section that has the exact outline of the tissue but no tissue. The tissue consistency could best be described as "dry?" Interestingly, it is only the normal livers that have this problem. We have tried sectioning thick, thin, no soak, cold soak......but out of ideas. Anyone have any suggestions? Unfortunately we have no control over the tissue processing, but perhaps can convince to change. Thanks Luis Happy Holidays to all!! _______________________________________________ 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 End of Histonet Digest, Vol 25, Issue 26 **************************************** Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From Malcolm.McCallum <@t> tamut.edu Fri Feb 10 18:10:34 2006 From: Malcolm.McCallum <@t> tamut.edu (Malcolm McCallum) Date: Fri Feb 10 18:12:09 2006 Subject: [Histonet] histology lab faculty startup Message-ID: Hi everyone, I am curious if there is a list out there somewhere of items new faculty should consider on their startup? Thanks for any help! Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Patsy Ruegg Sent: Fri 2/10/2006 3:10 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Mer2 antibody Has anyone tried Mer (N-19) sc6872 on ffpe tissues? This is UFO or ARK family of receptor tyrosine kinases also designated as Mer, Nyk or Eyk. Mer may be involved in the development of glioblastomas so we are using it on human brain tissue ffpe. I asked this question of the IHC Resource Group www.ihcrg.org but got no responses, so I thought I would put it out to histonet. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From laurie.reilly <@t> jcu.edu.au Fri Feb 10 23:33:20 2006 From: laurie.reilly <@t> jcu.edu.au (Laurie Reilly) Date: Fri Feb 10 23:33:24 2006 Subject: [Histonet] Does anyone have any suggestions for getting brain tissue (human) to stay on the slides? In-Reply-To: Message-ID: <5.1.0.14.0.20060211152128.03820db8@mail.jcu.edu.au> Dear Barb and Histonetters, Brain tissue does have a tendency to wash off the slide. This does seem to vary with the species - human and sheep have given me the most angst. We sub the slides with glycerine-albumin for all our brain sections. Then we dry the sections in an oven at 37C for many hours, preferably overnight. (our usual practice for other sections, including brain from other species, is to dry at 60C for 30 minutes) Hope this helps. Regards, Laurie. At 11:34 AM 10/02/2006 -0500, Barb Nuernberger wrote: >Does anyone have any suggestions for getting brain tissue (human) to >stay on the slides? >Fixation? > >Barb > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mr.Laurie Reilly Ph 07 4781 4468 School of Veterinary & Biomedical Science Fax 07 4779 1526 James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. From vanann702 <@t> skmc.gov.ae Sat Feb 11 01:08:53 2006 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Sat Feb 11 01:08:05 2006 Subject: [Histonet] unsubscriube Message-ID: <7E070A7B959A9F42BE732545E5CF6210016F4C45@SKMCEMAIL.skmc.gov.ae> From vanann702 <@t> skmc.gov.ae Sat Feb 11 01:45:17 2006 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Sat Feb 11 01:44:26 2006 Subject: [Histonet] unsubscribe Message-ID: <7E070A7B959A9F42BE732545E5CF6210016F4CEF@SKMCEMAIL.skmc.gov.ae> From amosbrooks <@t> gmail.com Sat Feb 11 11:11:46 2006 From: amosbrooks <@t> gmail.com (Amos Brooks) Date: Sat Feb 11 11:11:58 2006 Subject: [Histonet] Let's go Histo (AKA changing the subject) Message-ID: <582736990602110911k5f0cb624xa55635f66d619fa7@mail.gmail.com> Hi, In case anyone plans on running with this idea , here is a site with the lyrics http://www.lyricsdemon.com/lyrics.php?songid=158855 for example: If u don't like the world you're living in Take a look around u At least u got friends becomes: If u don't like the lab you're workin' in Take a look around u At least u got stains ... Could be fun ... could be terrible! One thing for sure it makes for more interesting subject matter than recruiter bashing :-) Amos ------------------------------ > > Message: 14 > Date: Thu, 9 Feb 2006 22:41:09 GMT > From: "mprice26@juno.com" > Subject: Re: [Histonet] RE: Claire Ingels' (re: Asking for your help > ...) > To: KevinMcGovern@catholichealth.net > Cc: histonet@lists.utsouthwestern.edu > Message-ID: <20060209.144154.29586.235078@webmail65.nyc.untd.com> > Content-Type: text/plain > > Histonetters, > I want everyone to vote on using Claire's phrase of "Go Histo" for the > 2007 NSH Phrase. We could redo Prince's song to say "Let's go Histo" insted > of "Let's Go Crazy". > > Marsha Price > > From JWEEMS <@t> sjha.org Sat Feb 11 18:21:46 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Sat Feb 11 18:23:27 2006 Subject: [Histonet] Does anyone have any suggestions for gettingbrain tissue (human) to stay on the slides? Message-ID: <83AACDB0810528418AA106F9AE9B7F7E018252A0@sjhaexc02.sjha.org> I've found that putting them on plus slides, air drying overnight, then drying in an oven the usual - 20-30 minutes works. I have only had experience with human. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Laurie Reilly Sent: Sat 2/11/2006 12:33 AM To: Barb Nuernberger; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Does anyone have any suggestions for gettingbrain tissue (human) to stay on the slides? Dear Barb and Histonetters, Brain tissue does have a tendency to wash off the slide. This does seem to vary with the species - human and sheep have given me the most angst. We sub the slides with glycerine-albumin for all our brain sections. Then we dry the sections in an oven at 37C for many hours, preferably overnight. (our usual practice for other sections, including brain from other species, is to dry at 60C for 30 minutes) Hope this helps. Regards, Laurie. At 11:34 AM 10/02/2006 -0500, Barb Nuernberger wrote: >Does anyone have any suggestions for getting brain tissue (human) to >stay on the slides? >Fixation? > >Barb > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mr.Laurie Reilly Ph 07 4781 4468 School of Veterinary & Biomedical Science Fax 07 4779 1526 James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From kemlo <@t> f2s.com Sun Feb 12 06:40:15 2006 From: kemlo <@t> f2s.com (kemlo) Date: Sun Feb 12 06:40:30 2006 Subject: [Histonet] RE: purple blue haze In-Reply-To: <5B3B26C4B71D5E469892D1860ABE10EA7F1E8D@SHSEXVS01.int.samhealth.net> Message-ID: <200602121240.k1CCeFVT032647@outmail.freedom2surf.net> Obviously something in the water! If you restain using hot water does the haze disappear? You must assume I suppose that something crystallises out of cold water and that the hot water is either 'cleaner' or is sufficiently warm to dissolve (or keep dissolved) what ever is doing it. In London we never blued in tap water, but then you ever did anything with London water but use it in the toilet. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of lharris@samhealth.org Sent: 10 February 2006 22:02 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: purple blue haze We had a purple blue haze on our H&E's when we used cold tap water run through the stainer. We had to use hot water and the haze went away. I've tried to return to using cold water and we get the haze every time. Lori A. Harris, HT (ASCP) Histology Section Leader GSRMC - Pathology 3600 NW Samaritan Drive Corvallis, OR 97330 1-541-768-6078 lharris@samhealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, December 21, 2005 8:06 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 25, 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-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@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. online journals (Malcolm McCallum) 2. Purple blue haze (Heckford, Karen - SMMC-SF) 3. Re: online journals (John A. Kiernan) 4. Re:Sections coming off! (Shannon Bryce) 5. re: MMA sections coming off! (Tracey Couse) 6. Re 17. fluorescence immunohistochemistry (Donna Harclerode) 7. Re: Purple blue haze (Andrea Grantham) 8. RE: Purple blue haze (Laurie Colbert) 9. Re: anti-human VEGF antibody (pruegg@ihctech.net) 10. Re: 17. fluorescence immunohistochemistry (John Kiernan) 11. Re: non-specific staining IHC (Malam Jacqueline) 12. MMA sections falling off (Nancy W. Troiano) 13. Modified Davidson's Rat Eyes and Testes Fixation Times (Sharon E Willman) 14. Embedding Centers (Roberta Horner) 15. Glass coverslippers (Angela Bitting) 16. RE: Glass coverslippers (Bartlett, Jeanine) 17. Re: Embedding Centers (Rene J Buesa) 18. San Antonio Histology connection (Jackie M O'Connor) 19. RE: Embedding Centers (Anne Van Binsbergen) 20. RE: Glass coverslippers (Anne Van Binsbergen) 21. Re: MMA sections falling off (Gayle Callis) 22. Re: Embedding Centers (Pamela Marcum) 23. decal rapid (Sharon Allen) 24. FW: artifact (Santana, Diane) 25. antibody search (Steven Coakley) 26. Mouse liver tissue processing (Luis Chiriboga) 27. RE: Mouse liver tissue processing (HSRL) ---------------------------------------------------------------------- Message: 1 Date: Tue, 20 Dec 2005 12:11:01 -0600 From: "Malcolm McCallum" Subject: [Histonet] online journals To: , , , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hi Which of the below three options do you think is best regarding a journal with both print and online versions???? 1) Journal is open access, the publisher will charge page charges to authors. 2) No page charges (except color plates), but the publisher charges download fees and only the abstracts are open access. 3) Page charges are optional for the author, if the author pays the entire article is open access, if not then only the abstract will be accessible without paying a download fee. In case someone is unaware, If an article is entirely open access then anyone can read it or download it online. IF only the abstract is available then you can read the abstract online, but must pay a fee to download the entire article. Thanks for the feedback, its actually very important! Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html ------------------------------ Message: 2 Date: Tue, 20 Dec 2005 11:17:21 -0700 From: "Heckford, Karen - SMMC-SF" Subject: [Histonet] Purple blue haze To: "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Dear Histonetters, Has anyone had any problems with a purple- blue haze on their H&E using Richard Allen Scientific Hematoxylin 7211. I am currently filtering it. I am still getting the haze. I have changed nothing, it just appeared a couple of weeks ago. Any help would be greatly appreciated. Happy Holidays, Karen Heckford HT (ASCP) CE Lead Histology Technician St. Mary's Medical Center Histology Department 450 Stanyan St. San Francisco, Ca. 94117 1-415-668-1000 ext. 6167 email: kheckfor@chw.edu ------------------------------ Message: 3 Date: Tue, 20 Dec 2005 13:42:34 -0500 From: "John A. Kiernan" Subject: Re: [Histonet] online journals To: Malcolm McCallum Cc: histonet@lists.utsouthwestern.edu, tws-l@list.wildlife.org, ecolog-l@listserv.umd.edu, parc@listserv.uga.edu Message-ID: <43A8509A.9B855C2A@uwo.ca> Content-Type: text/plain; charset=us-ascii You raise some interesting points. Clearly you're polling a wide range of scientists; please send out a summary when you've analysed the results. Here's my two-centsworth. Paying to read the full text doesn't apply if your institution subscribes to the journal (or more usually to a large batch from the publisher). Since most people who want the full text are in institutions with libraries (or with employers such as drug companies that will pay for individual articles), avoiding page charges is the way to go. Why should the researcher and author pay the publisher? It should be the other way round. Prestigious journals commonly do charge authors, but anyone publishing in such a journal is sure to have a big enough research grant to cover the fees, which are small compared with salaries and items for the lab. Some journals published by learned societies do not charge authors anything, even for coloured pictures if these are needed to make a point. Three examples that come to mind are the Journal of Anatomy, the Journal of Histochemistry and Cytochemistry, and Biotechnic & Histochemistry. I'm sure there are many others. All three that I mentioned come free with membership in the society. The cost works out at about $5 per issue (print, and access online). I've not heard of the option of the author paying and the reader not paying. It's hard to see that working; even non-profit societies cannot run their journals at a loss. Libraries have to pay much more than individual members. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Malcolm McCallum wrote: > > Hi > Which of the below three options do you think is best regarding a journal with both print and online versions???? > > 1) Journal is open access, the publisher will charge page charges to authors. > 2) No page charges (except color plates), but the publisher charges download fees and only the abstracts are open access. > 3) Page charges are optional for the author, if the author pays the entire article is open access, if not then only the abstract will be accessible without paying a download fee. > > In case someone is unaware, If an article is entirely open access then anyone can read it or download it online. IF only the abstract is available then you can read the abstract online, but must pay a fee to download the entire article. > > Thanks for the feedback, its actually very important! > > Malcolm L. McCallum > Assistant Professor > Department of Biological Sciences > Texas A&M University Texarkana > 2600 Robison Rd. > Texarkana, TX 75501 > O: 1-903-233-3134 > H: 1-903-791-3843 > Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Tue, 20 Dec 2005 13:01:17 -0600 From: "Shannon Bryce" Subject: [Histonet] Re:Sections coming off! To: "Joanne Mauger" , , Message-ID: <43A8009D0200009900000BC0@swnw124.swmed.edu> Content-Type: text/plain; charset=US-ASCII We get our slides which are the + coated slides from Fisher. Our MMA sections are also falling off here lately so I went to another department and borrowed a box of theirs that they bought through VWR and have had no problems so I would guess it's a bad lot.....anyone have lot numbers? Thanks, Shannon Bryce UT Southwestern Medical Center >>> "Joanne Mauger" 12/20/05 8:39 AM >>> Hi Patricia, Are you using slides from Erie? We buy ours through Fisher, and have been having trouble keeping sections on + slides as well. It is as if they have no charge at all. We think the lot may be bad. The company claims to have no complaints. Anyone else having similar problem? Jo Mauger >>> "Patricia F Lott" 12/19/05 3:21 PM >>> Help! We are losing our thin plastic sections! We have cut PMMA plastic sections at 5 microns and put them on "+" slides, and the sections fall off either during de-plasticizing or during staining! We have tried: Haupt's adhesive, longer drying times, "+" slides, plus slides and Haupt's together, etc....no luck! Any suggestions would be welcome! _______________________________________________ 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 ------------------------------ Message: 5 Date: Tue, 20 Dec 2005 15:15:42 -0500 From: Tracey Couse Subject: re: [Histonet] MMA sections coming off! To: histonet@lists.utsouthwestern.edu Message-ID: <43A8666E.2030005@ibb.gatech.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Patricia, Try manually coating plain glass slides with poly-l-lysine, chrome/gelatin, or glycerin/gelatin. We use all of these for our thin plastic sections. I also suggest setioning a micron or two thinner (cut at 3 or 4um). A thinner section will reduce the surface area to volume ratio and may promote better section adherance. You may need to dry the sldies using a slide press and dry longer or with higher temperatures. Also, try searching the Histonet archives for alternative slide coating suggestions. Good luck! -- Tracey Couse Laboratory Coordinator Georgia Tech/Emory Center for the Engineering of Living Tissues Georgia Institute of Technology IBB, Room 1123 Office: 404.385.2611 Lab: 404-385-6735 Fax: 404.894.229 Date: Mon, 19 Dec 2005 14:21:07 -0600 From: "Patricia F Lott" Subject: [Histonet] MMA sections coming off! To: Message-ID: Content-Type: text/plain; charset="us-ascii" Help! We are losing our thin plastic sections! We have cut PMMA plastic sections at 5 microns and put them on "+" slides, and the sections fall off either during de-plasticizing or during staining! We have tried: Haupt's adhesive, longer drying times, "+" slides, plus slides and Haupt's together, etc....no luck! Any suggestions would be welcome! 1 ------------------------------ Message: 6 Date: Tue, 20 Dec 2005 13:06:17 -0800 From: "Donna Harclerode" Subject: [Histonet] Re 17. fluorescence immunohistochemistry To: , Cc: Adam Johnson Message-ID: <3DE0F644E093DF4BAE80C254176696A505B869@mp-mailserver.macropore.com> Content-Type: text/plain; charset="US-ASCII" Message: 17 Date: Mon, 19 Dec 2005 22:08:48 +0100 From: Emily.Wiesner@medecine.unige.ch Subject: [Histonet] fluorescence immunohistochemistry To: histonet@lists.utsouthwestern.edu Message-ID: <1135026528.43a72160c086f@webmail.medecine.unige.ch> Content-Type: text/plain; charset=ISO-8859-1 Hi All, I am new to fluorescence immunohistochemistry and I was wondering if anyone can let me know the simple steps involved in performing this technique. I know the dilutions required for the primary and secondary antibodies, I just need to known what solutions are used for washes and the steps in between! Thanks in advance, Emily Dear Emily I do very simple IHC using fluorescent secondaries from Jackson Immunoresearch and purified primary abs. Depending on what you are doing, there can be more steps added. I usually am staining sections on a slide or cells on chamber slides or in a plate. For the unfixed frozen sections on slides I normally would fix 5 min in 75% acetone 25% ethanol and then right into PBS- do not allow the fixed slides to dry. The only change would be if the antibody prefers formalin fixation to acetone/ alcohol (thanks Gayle- I love this fixative for almost everything). I load them into a Sequenza(tm) rack (Thermo Electron, formerly Shandon) or you could put them in a humid chamber of some sort, wiping excess buffer before adding antibody or blocking solutions. OPTIONAL I have recently started using Image-iT(tm) FX signal enhancer, I36933, $80 Invitrogen for auto fluorescence (not sure it helps yet, but sure does not hurt) 30 minutes. I would not use this on routinely- my sections have infracted areas that have been a problem in the past) After blocking, I rinse 1 time in the Sequenza rack (3 x 5 minutes if you are doing them by hand) Add 100ul of the antibody diluted in Dako antibody diluent S0809- usually 1:100- 1:1k (dilutions are antibody dependant, but I have found in fluorescence too high a concentration is not as big a problem as with HRP formats) If you are using a humid chamber be sure to cover all the tissue. I have incubated slides at Room temperature for 1-2 hours, but now am staining overnight in the fridge (not so good in clinical situations) After primary incubation, I rinse 1 time in the Sequenza rack (3 x 5 minutes if you are doing them by hand) Next is Jackson secondary at a 1:100 dilution (I use Cy3 conjugates for single color and FITC and RITC for dual color. All my secondaries are minimal cross (mouse to rat), but if all your tissue is human that is not necessary. 30 minutes I add DAPI to my secondaries to give a blue nuclear stain so I can tell what I am looking at and localize the stain. Rinse again 3 times in PBS and coverslip with your choice of aqueous media. For media that never gets hard, I prefer Immunomount (Thermo Electron, formerly Shandon) or Aqua Mount (Lerner from VWR) same thing different label or the Prolonge Mounting media P36930 for $101 that will harden. I do not like the hard mount media from Vector- I got many air bubbles forming day after the sldes were coverslipped. For cells on plates or chamber slides I usually fix with 10% NBF or 4% PFA (same concentration of fixative, 4%PFA will contain no methanol) for 5-30 minutes. CD makers can be very hard to stain with NBF but with a short fix, I have been pretty luck so far. I rinse 3 x 10 minutes with PBS I have not used the blocking reagents on cells as I have not seen any autofluorescence in our cardiac myocytes, neurons or stem cells. The rest of the procedure is the same except in a 6 well plate you need a lot of antibody to cover the plate (about 600u). I try to use a rocker if it is available to help to be sure of good coverage. I do not usually cover my slides or plates, but if your room is very bright or your signal very week, it would be a good idea to protect the secondaries from light. I like to use isotype controls (negative controls) or rabbit IgGs for much of my work, but in a pinch I use just the antibody diluent to be sure the stain I see, is a result of the primary antibody and not the secondary binding to some part of the cell. In cell assays I always want to stain a known positive and a known negative cell if at all possible in addition to the isotype controls. If you are making your own diluent (to me Dako diluent is great and I have used it successfully for over 10 years in many applications) I use 0.3% Triton, 5 % Normal serum from the species that your secondary is made in - (all my secondaries are made in donkey so I would use donkey serum) in PBS. I have used donkey secondaries when possible because when I tested them years ago, they were a bit cleaner on the CD markers I was using. Dako diluent must have a similar make up, but it lasts a year in the fridge and it permeablizes intact cells very nicely. One big thing when purchasing fluorophores, make sure the filters on your scope can see the secondaries you are purchasing. I found out at my current job after staining with Cy3 and RITC that the reason I could not get any stain was they had a Texas red filter not the Cy3, RITC filter. Ideally if you are doing multicolor you want narrow band filters so you can separate the colors, with broad pass filters the colors blur together and it is hard to separate the results. Good Luck! Donna Donna Harclerode, HT, (ASCP), HTL, QIHC Scientist / Immunohistochemistry Cytori Therapeutics 3020 Callan Rd. San Diego, CA 92121 858-458-0900 ext 322 dharclerode@cytoritx.com ------------------------------ Message: 7 Date: Tue, 20 Dec 2005 14:19:47 -0700 From: Andrea Grantham Subject: Re: [Histonet] Purple blue haze To: "Histonet (E-mail)" Message-ID: <4.3.2.7.2.20051220140523.00ce4520@algranth.inbox.email.arizona.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Karen, Yes I have recently noticed the "haze" of the H&E stained slides. I have been using this hematoxylin for years and this is the first time that I have had a problem with it. Like you I have filtered it but it hasn't seemed to help. I also have been using their Type 6 paraffin and in the last few months have noticed that it blows apart on the waterbath. I have lowered the temp. but it didn't help -> 35-39 degrees C. A different lot number that they sent to me was only marginally better. A couple of weeks ago one of my clients brought a project consisting of blocks that had been processed and embedded here over 4-5 years. The paraffin throughout was RAS Type 6. The blocks from the first 4 years cut fine and laid out on the waterbath fine but when I got to recently embedded blocks they started the rapid expanding and blowing apart when they were placed on the waterbath. A histotech in CA told me they have been having the same problem in their lab. Hmmmmm.....could they have changed the formula? Maybe this is so with the Hematoxylin as well. I'm trying different paraffins looking for one comparable to the Type 6 only more stable on the waterbath. I really don't want to do this with Hematoxylin because my clients love my staining but I hate ugly slides. Andi At 11:17 AM 12/20/2005 -0700, Heckford, Karen - SMMC-SF wrote: >Dear Histonetters, >Has anyone had any problems with a purple- blue haze on their H&E using >Richard Allen Scientific Hematoxylin 7211. I am currently filtering it. I >am still getting the haze. I have changed nothing, it just appeared a >couple of weeks ago. Any help would be greatly appreciated. > >Happy Holidays, > > >Karen Heckford HT (ASCP) CE >Lead Histology Technician >St. Mary's Medical Center >Histology Department >450 Stanyan St. >San Francisco, Ca. 94117 >1-415-668-1000 ext. 6167 >email: kheckfor@chw.edu > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html ------------------------------ Message: 8 Date: Tue, 20 Dec 2005 15:12:17 -0800 From: "Laurie Colbert" Subject: RE: [Histonet] Purple blue haze To: "Andrea Grantham" , "Histonet (E-mail)" Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628006307C76@EXCHANGE1.huntingtonhospital.com> Content-Type: text/plain; charset="iso-8859-1" Your discussion rang a bell with me, and I've been meaning to call Richard Allan. We use their Type 9 paraffin, and for the last 3 or 4 months it also has been "blowing apart on the waterbath." We lowered the temp on the waterbaths, and this seems to have helped. Our housekeeping personnel have even noticed a difference in the paraffin (scraping it off the floor). They say it is stickier lately. I think something has been changed. Laurie Colbert Huntington Hospital Pasadena, CA -----Original Message----- From: Andrea Grantham [mailto:algranth@u.arizona.edu] Sent: Tuesday, December 20, 2005 1:20 PM To: Histonet (E-mail) Subject: Re: [Histonet] Purple blue haze Karen, Yes I have recently noticed the "haze" of the H&E stained slides. I have been using this hematoxylin for years and this is the first time that I have had a problem with it. Like you I have filtered it but it hasn't seemed to help. I also have been using their Type 6 paraffin and in the last few months have noticed that it blows apart on the waterbath. I have lowered the temp. but it didn't help -> 35-39 degrees C. A different lot number that they sent to me was only marginally better. A couple of weeks ago one of my clients brought a project consisting of blocks that had been processed and embedded here over 4-5 years. The paraffin throughout was RAS Type 6. The blocks from the first 4 years cut fine and laid out on the waterbath fine but when I got to recently embedded blocks they started the rapid expanding and blowing apart when they were placed on the waterbath. A histotech in CA told me they have been having the same problem in their lab. Hmmmmm.....could they have changed the formula? Maybe this is so with the Hematoxylin as well. I'm trying different paraffins looking for one comparable to the Type 6 only more stable on the waterbath. I really don't want to do this with Hematoxylin because my clients love my staining but I hate ugly slides. Andi At 11:17 AM 12/20/2005 -0700, Heckford, Karen - SMMC-SF wrote: >Dear Histonetters, >Has anyone had any problems with a purple- blue haze on their H&E using >Richard Allen Scientific Hematoxylin 7211. I am currently filtering it. I >am still getting the haze. I have changed nothing, it just appeared a >couple of weeks ago. Any help would be greatly appreciated. > >Happy Holidays, > > >Karen Heckford HT (ASCP) CE >Lead Histology Technician >St. Mary's Medical Center >Histology Department >450 Stanyan St. >San Francisco, Ca. 94117 >1-415-668-1000 ext. 6167 >email: kheckfor@chw.edu > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Tue, 20 Dec 2005 17:56:17 -0800 (PST) From: pruegg@ihctech.net Subject: Re: [Histonet] anti-human VEGF antibody To: "Katri Tuomala" Cc: Histonet Message-ID: <43434.207.200.116.10.1135130177.squirrel@207.200.116.10> Content-Type: text/plain;charset=iso-8859-1 I am using mouse anti-vegf from Santa Cruz with good results on human and baboon ffpe tissues, it is not easy, but I do HIER in a water bath in high ph buffer at 85dc for 3 hrs., followed by overnight incubation of the primary 1:200, detected with mouse labeled polymer (I use Envision from DAKO). Patsy > Andrea, > I cannot say this is the best anti-VEGF antibody, but I am doing a > research > project on FFPE human brain tissue and using placenta as a control with > following antibody: > monoclonal mouse anti human VEGF, clone VG1 from ID Labs (www.idlabs.com), > dilution 1:100, HIER in citrate buffer pH 6.0. Detection system is from > Zymed, Histostain Plus Kit.. > Glioblastoma stains strongly with this antibody. I have not used this with > frozen sections. > > Katri > > Katri Tuomala > Hamilton, Ontario, Canada > ----- Original Message ----- > From: "Andrea T. Hooper" > To: "Histonet" > Sent: Monday, December 19, 2005 8:37 PM > Subject: [Histonet] anti-human VEGF antibody > > >> What's the current consensus on the best antibody to use for anti-human >> VEGF immunostaining on human FFPE sections (frozens as well if >> possible). >> >> Thanks so much!! >> Andrea >> -- >> >> _______________________________________________ >> 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 > ------------------------------ Message: 10 Date: Wed, 21 Dec 2005 02:10:48 -0500 From: John Kiernan Subject: Re: [Histonet] 17. fluorescence immunohistochemistry To: Donna Harclerode Cc: histonet@lists.utsouthwestern.edu, Adam Johnson , Emily.Wiesner@medecine.unige.ch Message-ID: <43A8FFF8.D988E7C6@uwo.ca> Content-Type: text/plain; charset=us-ascii When I was a student in Birmingham, England (that's the Birmingham with the silent h), the immunology and serology lecturer told us: "water is unphysiological and beer is expensive, so we use saline." He must have been a good lecturer because I can still recall that apophthegm 40 years on when afar and asunder! John Kiernan London, Ontario. ------------------------------- > Hi All, > I am new to fluorescence immunohistochemistry and I was wondering if > anyone can let me know the simple steps involved in performing this > technique. I know the dilutions required for the primary and secondary > antibodies, I just need to known what solutions are used for washes and > the steps in between! > Thanks in advance, ------------------------------ Message: 11 Date: Wed, 21 Dec 2005 12:00:58 -0000 From: Malam Jacqueline Subject: [Histonet] Re: non-specific staining IHC To: "Histonet submissions (histonet@lists.utsouthwestern.edu)" Message-ID: Content-Type: text/plain We use the Benchmark XT but make our own prep kits up for neg controls (poly and mono) and are not experiencing any more non-sp staining than we normally do which is minimal. Jacqui DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. ------------------------------ Message: 12 Date: Wed, 21 Dec 2005 07:20:53 -0500 From: "Nancy W. Troiano" Subject: [Histonet] MMA sections falling off To: histonet@lists.utsouthwestern.edu Message-ID: <5.2.1.1.2.20051221071248.00c2c038@email.med.yale.edu> Content-Type: text/plain; format=flowed; charset=us-ascii We use Chrome-alum gel coated slides and prepare them as follows: dissolve 9 g gelatin(Fisher #G8-500) per 1000 ml distilled water (solution A) while gently heating solution. Dissolve 4 g Chromium Potassium Sulfate (Fisher #C337-500) in distilled water (solution B). Prepare gelatin solution for slide dip: Mix 500 ml Solution a with 19.25 ml solution B. Heat to 70-75 Deg. C, dip slides 2 -4 minutes in this solution. Allow to dry vertically, then put in hot oven overnight (60 deg. C). To adhere sections to slides we place the 5 micron MMA sections on slide, spread with 70% ethanol, cover with a piece of plastic and stack slides then clamp with an office clamp. Put slides in 37deg C oven for two nights or for one night in 60 deg C oven (don't do the hot oven if you are subsequently staining for enzymes). For immuno we mount our 5 micron plastic sections on hybridization slides from Scientific Device Laboratory, Cat. #063 and place in hot oven 60 deg C overnite. Hope this helps! ------------------------------ Message: 13 Date: Wed, 21 Dec 2005 07:18:32 -0600 From: Sharon E Willman Subject: [Histonet] Modified Davidson's Rat Eyes and Testes Fixation Times To: histonet@lists.utsouthwestern.edu Message-ID: <43A95628.7060609@bms.com> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Hi, I was needing input on how long you fix rat eyes and testes in Modified Davidson's. Any suggestions would be most appreciated. Thanks in advance, Sharon Willman ------------------------------ Message: 14 Date: Wed, 21 Dec 2005 08:25:26 -0500 From: "Roberta Horner" Subject: [Histonet] Embedding Centers To: Message-ID: Content-Type: text/plain; charset="US-ASCII" My embedding center just died. Last time our maintenance looked at it they couldn't get the parts (its about 17 years old). I need to get information quick on which ones you like and are good. So far I have quotes from Surgipath and on the Leica but I don't know anything about them. Any comments please? Roberta ------------------------------ Message: 15 Date: Wed, 21 Dec 2005 08:49:01 -0500 From: "Angela Bitting" Subject: [Histonet] Glass coverslippers To: Message-ID: Content-Type: text/plain; charset=US-ASCII Can anyone recommend a automated glass coverslipper? I've used Hacker in the past. Any that are faster, better? Thanks. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. ------------------------------ Message: 16 Date: Wed, 21 Dec 2005 08:56:05 -0500 From: "Bartlett, Jeanine" Subject: RE: [Histonet] Glass coverslippers To: "Angela Bitting" , Message-ID: Content-Type: text/plain; charset="us-ascii" We recently demo'd the new one from Leica and it was great! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Angela Bitting Sent: Wednesday, December 21, 2005 8:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Glass coverslippers Can anyone recommend a automated glass coverslipper? I've used Hacker in the past. Any that are faster, better? Thanks. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 17 Date: Wed, 21 Dec 2005 06:12:16 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Embedding Centers To: Roberta Horner , histonet@lists.utsouthwestern.edu Message-ID: <20051221141216.20482.qmail@web61215.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi Roberta: I would suggest you to call Sakura-Finetek. They have a new embedding center that has all the advantages of the old Tissue-Tek, along with the Sakura reliability. Ask for a "demo" instrument (they usually accomodate that type of request). Ren? J. Roberta Horner wrote: My embedding center just died. Last time our maintenance looked at it they couldn't get the parts (its about 17 years old). I need to get information quick on which ones you like and are good. So far I have quotes from Surgipath and on the Leica but I don't know anything about them. Any comments please? Roberta _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 18 Date: Wed, 21 Dec 2005 08:25:29 -0600 From: "Jackie M O'Connor" Subject: [Histonet] San Antonio Histology connection To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Could someone in the San Antonio area contact me directly? Thanks. Jackie O' ------------------------------ Message: 19 Date: Wed, 21 Dec 2005 18:20:37 +0400 From: "Anne Van Binsbergen" Subject: RE: [Histonet] Embedding Centers To: "Roberta Horner" , Message-ID: <7E070A7B959A9F42BE732545E5CF621094ACBA@SKMCEMAIL.skmc.gov.ae> Content-Type: text/plain; charset="us-ascii" Im a fan of the Sakura embedding system - it's ergonomically designed and VERY user friendly Can be 'left handed' or 'right handed' Cassettes left/right Moulds left/right Cold plate left/right Forceps left/right Nothing more annoying (being a lefty) to be forced to work like a non-lefty!! Everything works independently of everything else - ie - micro adjustments can be made to temp in almost any area to your requirements Nice broad anti burn protection to rest hands/fingers while embedding And if you have a VIP5 the basket fits snugly into the holding area - left or right as you prefer of course!! Like I said - I am THE fan of Sakura And I just KNOW that some of you are smiling out there - you know who you are!!! Annieinarabia -----Original Message----- From: Roberta Horner [mailto:rjr6@psu.edu] Sent: Wednesday, December 21, 2005 5:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Embedding Centers My embedding center just died. Last time our maintenance looked at it they couldn't get the parts (its about 17 years old). I need to get information quick on which ones you like and are good. So far I have quotes from Surgipath and on the Leica but I don't know anything about them. Any comments please? Roberta _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 20 Date: Wed, 21 Dec 2005 18:24:12 +0400 From: "Anne Van Binsbergen" Subject: RE: [Histonet] Glass coverslippers To: "Bartlett, Jeanine" , "Angela Bitting" , Message-ID: <7E070A7B959A9F42BE732545E5CF6210948C5F@SKMCEMAIL.skmc.gov.ae> Content-Type: text/plain; charset="us-ascii" Try the Sakura Fast, efficient, small 'footprint' - will fit into a small space Had a Leica once - was so very patient - 2 exchanges (new machines) later - still hated it Annieinarabia -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: Wednesday, December 21, 2005 5:56 PM To: Angela Bitting; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Glass coverslippers We recently demo'd the new one from Leica and it was great! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Angela Bitting Sent: Wednesday, December 21, 2005 8:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Glass coverslippers Can anyone recommend a automated glass coverslipper? I've used Hacker in the past. Any that are faster, better? Thanks. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ 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 ------------------------------ Message: 21 Date: Wed, 21 Dec 2005 08:14:11 -0700 From: Gayle Callis Subject: [Histonet] Re: MMA sections falling off To: "Nancy W. Troiano" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20051221081017.01b39c08@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed To increase the holding power, the gelatin can be 275 bloom, a larger gelatin molecule made from pig collagen. This subbing solution suggested by Nancy is tried and true, also works for the nasty decalcified bone sections, generally larger ones. However, if the subbing solution causes too much blue background staining from hematoxylin, one can dip the subbed slides in NBF a couple of times, rinse well, dry and store in cool, dry place. The NBF crosslinks the gelatin a bit and reduces blue background. At 05:20 AM 12/21/2005, you wrote: >We use Chrome-alum gel coated slides and prepare them as >follows: dissolve 9 g gelatin(Fisher #G8-500) per 1000 ml distilled >water (solution A) while gently heating solution. Dissolve 4 g Chromium >Potassium Sulfate (Fisher #C337-500) in distilled water (solution >B). Prepare gelatin solution for slide dip: Mix 500 ml Solution a with >19.25 ml solution B. Heat to 70-75 Deg. C, dip slides 2 -4 minutes in >this solution. Allow to dry vertically, then put in hot oven overnight >(60 deg. C). To adhere sections to slides we place the 5 micron MMA >sections on slide, spread with 70% ethanol, cover with a piece of plastic >and stack slides then clamp with an office clamp. Put slides in 37deg C >oven for two nights or for one night in 60 deg C oven (don't do the hot >oven if you are subsequently staining for enzymes). For immuno we mount >our 5 micron plastic sections on hybridization slides from Scientific >Device Laboratory, Cat. #063 and place in hot oven 60 deg C >overnite. Hope this helps! > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 22 Date: Wed, 21 Dec 2005 10:14:58 -0500 From: Pamela Marcum Subject: Re: [Histonet] Embedding Centers To: "Roberta Horner" , Message-ID: <6.1.1.1.2.20051221101125.01997318@mail.vet.upenn.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed At 08:25 AM 12/21/2005, Roberta Horner wrote: >My embedding center just died. Last time our maintenance looked at it >they couldn't get the parts (its about 17 years old). I need to get >information quick on which ones you like and are good. So far I have >quotes from Surgipath and on the Leica but I don't know anything about >them. Any comments please? >Roberta >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Hi, I will be totally different as i am buying the Thermo Shandon Embedding Center. I had it in for demo and it had all the features the others have for timing and pre-sets as well as some independent temperature ranges I likes. Also the cold plate is seperate so I can have it on either side or in another area if I want for special projects. Good Luck, I think they all work well it just depends for me on features and price and Shandon also had the right price. Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu ------------------------------ Message: 23 Date: Wed, 21 Dec 2005 09:17:14 -0600 From: Sharon Allen Subject: [Histonet] decal rapid To: "Histonet (E-mail)" Message-ID: <36FE435D6D2F1D489174B22362A961B66B830B@hscxntmx0006.hsc.mb.ca> Content-Type: text/plain; charset="iso-8859-1" Hi, Does anyone know about or what supplier "Decal Rapid" by "Pathtech" can be purchased from? We had used it a few years ago but have no idea where we got it from. I tried searching the internet & came up with Pathtech in Australia but would like a supplier a little closer. Thanks in advance Sharon sallen@hsc.mb.ca This e-mail and/or any documents in this transmission is intended for the address(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. ------------------------------ Message: 24 Date: Wed, 21 Dec 2005 10:27:06 -0500 From: "Santana, Diane" Subject: [Histonet] FW: artifact To: "'Histonet@lists.utsouthwestern.edu'" Message-ID: <4C96AA62BADFD81180CB0002A5AD67FB04D7E282@MAILPMA> Content-Type: text/plain; charset="iso-8859-1" > -----Original Message----- > From: Santana, Diane > Sent: Tuesday, December 20, 2005 10:20 AM > To: 'histonet@lists.utsouthwestern.edu' > Subject: artifact > > I am having a problem with artifact on some of my GI bx's. Actually a > paper published by Anatech "The Innovator", summer2004 shows this same > artifact on the front page. They believe it is a fixation problem, and or > retrieval by the endo Drs. My pathologist rules both these ideas out. I > tend to agree with him, at lease on the fixation. Our bx's go into the > processor around 5 pm and the bx run starts around 2 am. > It is a loss of nuclear detail. This artifact can be on a small section of > a bx. Even if there is 5 bx's in one cassette maybe 1 will have this > artifact where the others look great. I thought at first it was a clearing > problem or nuclear bubbling I changed times in clear rite and oven. It did > not eliminate the problem. My pathologist has seen this by other labs > also, but very seldom. For us it could be 1 or 2 slides a day. Sometimes > not at all. Weekend or weekdays. No consistently. The best way I can > describe it is that it looks like a smudge. > Our bx run is this; > > 10% NBF 30 min ea x2 > 80% Alcohol 10mins > 95% alcohol 10 mins ea x2 > 100% alcohol 17 mins ea x3 > Clear rite 22 mins ea x2 > Paraffin 15 mins ea x3 > > I would appreciate any feedback on this. > Diane Santana > Pentucket Medical Assoc. > Haverhill, Mass. ------------------------------ Message: 25 Date: Wed, 21 Dec 2005 07:22:41 -0800 (PST) From: Steven Coakley Subject: [Histonet] antibody search To: Histonet@lists.utsouthwestern.edu Message-ID: <20051221152241.11885.qmail@web90204.mail.scd.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Gooday everyone, I'd like to get a few recommendations for anti-insulin and anti-glucagon antibodies for FFPE mouse tissue. If anyone has used a specific companies antibodies that work well please let me know. I'm still learning this research IHC stuff so I can use all the help I can "muster" Steve __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 26 Date: Wed, 21 Dec 2005 09:54:28 -0500 From: Luis Chiriboga Subject: [Histonet] Mouse liver tissue processing To: Histonet Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hi everyone We are having a real serious problem getting good sections from mouse livers. The tissue curls and shreds out of the block as soon as it comes in contact with the knife, end up with a section that has the exact outline of the tissue but no tissue. The tissue consistency could best be described as "dry?" Interestingly, it is only the normal livers that have this problem. We have tried sectioning thick, thin, no soak, cold soak......but out of ideas. Anyone have any suggestions? Unfortunately we have no control over the tissue processing, but perhaps can convince to change. Thanks Luis Happy Holidays to all!! ------------------------------ Message: 27 Date: Wed, 21 Dec 2005 11:02:33 -0500 From: "HSRL" Subject: RE: [Histonet] Mouse liver tissue processing To: "'Luis Chiriboga'" , Message-ID: <000c01c60647$f47a7ab0$0200a8c0@HSRLMAIN> Content-Type: text/plain; charset="us-ascii" Luis, Have a tried keeping them on a WET ice for 45 minutes to an hour? If you have tried it and it doesn't work, it is a processing problem that you must rectify. -Tom Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 5930 Main Street Mount Jackson, Virginia 22842 540.477.4440 540.477.4448 tomgalati@hsrl.org www.hsrl.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Luis Chiriboga Sent: Wednesday, December 21, 2005 9:54 AM To: Histonet Subject: [Histonet] Mouse liver tissue processing Hi everyone We are having a real serious problem getting good sections from mouse livers. The tissue curls and shreds out of the block as soon as it comes in contact with the knife, end up with a section that has the exact outline of the tissue but no tissue. The tissue consistency could best be described as "dry?" Interestingly, it is only the normal livers that have this problem. We have tried sectioning thick, thin, no soak, cold soak......but out of ideas. Anyone have any suggestions? Unfortunately we have no control over the tissue processing, but perhaps can convince to change. Thanks Luis Happy Holidays to all!! _______________________________________________ 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 End of Histonet Digest, Vol 25, Issue 26 **************************************** Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From esther.peters <@t> verizon.net Sun Feb 12 14:23:19 2006 From: esther.peters <@t> verizon.net (Esther Peters) Date: Sun Feb 12 14:23:20 2006 Subject: [Histonet] Learning histology References: <8C7FC9904048E67-FC4-9C6@FWM-D44.sysops.aol.com> Message-ID: <43EF9937.2020003@verizon.net> Stephen has received excellent advice from several folks on learning histology. Note that the emphasis has to be on "learning" through individual study done by the student, with as much time spent at the microscope or looking at online histology study guides as possible, learning the terminology and key features of the cells and tissues. This should include learning to think about what you are learning. In histology, structure relates to function! Think about why you might find goblet cells in one part of the digestive tract and not in another, what makes striated muscle useful in one place and smooth muscle in another, etc. I found with my students that thinking was often very difficult (as was studying on their own)! So read your histology textbook and atlas carefully, understand functions, think about the identifying features, and look at as many different examples of the same tissues as you can! Esther Peters, Ph.D. George Mason University mtitford@aol.com wrote: > Stephen at Texas A & M wants to know how to learn histology. I think the best way is to borrow a set of teaching microscope slides from your anatomy department (or cellular biology, whatever they call it now) and sit down with a microscope. > > 1) First of all study the different types of cells ( three types of muscle cells, the connective tissue cells, blood cells and developing cells, all the epithelial cells, glandular cells, etc) > 2) Then study the organ systems (Kidney, lung, intestine, nervous system, G.I. Tract, endocrine etc).Learn to diffentiate the different parts of the stomach, large and small intestines, brain, etc. Nearly every organ has one or two key structures that "give it away". Try to learn those. > 3) Then learn the tricks professors play on you: be able to differentiate resting breast from prostate, lactating breast from thyroid, spleen from lymph node, etc. > This is a time consuming method but it pays off. Modern histology classes are often taught using powerpoint or some other method where they use the best d*** photographs they can find of that organ or structure. In real life you have to look around the slide, orient yourself, find identfying features in the tissue, and then name it! Career histotechnologists add an additional step of learning the special stains associated with each particular organ or tissue. > > Good luck with your studies > > Mike Titford > USA Pathology > Mobile AL USA > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From lmt2104 <@t> columbia.edu Sun Feb 12 15:52:28 2006 From: lmt2104 <@t> columbia.edu (Lauren Tanabe) Date: Sun Feb 12 15:54:51 2006 Subject: [Histonet] Immunohistochemistry on brain: free floating versus cryostat mounted sections Message-ID: <671e238057fcd4dadbed7b2a8ef28dae@columbia.edu> Hi. I'm new to histonet and have found it very helpful. I was hoping to get some feedback on the following. I'm currently trying to figure out the best way to do DAB staining on mouse brain from a variety of ages ranging from E 15 up throuh P 14. If I try to do free floating sections on the younger animals, the sections fall apart. At the same time, I just can't get my cryostat sections to look nice. I've been perfusing with 4% PFA with a 3 hr post fix. I'm thinking that this is probably not long enough. I'm starting from scratch here so I'm trying to figure out: 1. The best way to slice and stain embryonic through P14 sections 2. The optimal thickness 3. If using cryostat mounted sections: How to get my sections to stick to the slides better. I invariably get little bits of brain that come up off the slide or maddening little bubbles underneath my section. (I'm using Fisher Superfrost/Plus slides) 4. The best way to mount free floating sections. The few times I have tried this, I get to the last step (with the help of mesh wells that fit into 12 well tissue culture plates) and when I pick it up with a brush I have a very difficult time transferring the section and not tearing it. --I also notice that I get little grid marks on my section from the mesh. Not terrible, but I'd like to eliminate it if possible. --Also the best way to minimize background. And when to do steps at room temperature versus at 4 degrees. Sorry if some of these questions are naive. I'm very new to the world of histology. Thanks in advance, Lauren Columbia University, NY From p.simpson <@t> improvision.com Mon Feb 13 02:52:51 2006 From: p.simpson <@t> improvision.com (Pippa Simpson) Date: Mon Feb 13 02:53:01 2006 Subject: [Histonet] unsubscribe References: <7E070A7B959A9F42BE732545E5CF6210016F4CEF@SKMCEMAIL.skmc.gov.ae> Message-ID: <013b01c6307a$df1c8750$0701a8c0@improvision.com> _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.m.vanderloos <@t> amc.uva.nl Mon Feb 13 03:14:20 2006 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Mon Feb 13 03:14:33 2006 Subject: [Histonet] RE: Mouse tissue staining with monoclonal mouse Abs. Message-ID:

Dear Guillermo,

Perhaps you should try a biotin-free detection system instead of t In my hands endogenous biotin cannot be effectively kidney tissue. Molecular Probes/Invitrogen has a Zenon kit in vitro labeling your mouse primary antibody with either enzymes fluorochromes via a labeled Fab anti-mouse reagent. Then c ontinue either with fluorescence microscopy or anti-FITC for enzymatic de van der Loos, PhD
Dept. of Pathology
Acade Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterd am
The Netherlands

 

Date: Fri, 10 Feb 2006 15:11:55 +0100 (CET)
From: <gpbnas@yahoo.es>
Subject: [ Mouse tissue staining with monoclonal mouse Abs.
Hel Vector Mouse on Mouse kit.
To: Histonet <histone t@lists.utsouthwestern.edu

Dear all,
&nbs with mouse frozen sections o implying an important mouse IgG kidney). For this reason we bought avoid detection of deposited IgG by seconda maintaining sensitivity for mouse primary antibody. increasing concentration of blocking reagent as well as decreasing concentration of biotynilated anti-mouse IgG, there still is a "back glomeruli togeth I would really appre helpful.
 &n advance,


Gui Ph.D.
Laboratorio de Reumatolog?a
Hospital 12 de Octubre
Avda de C?rdoba s/n
Madrid 28041
Spain

From TMcNemar <@t> lmhealth.org Mon Feb 13 04:17:30 2006 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Mon Feb 13 04:15:15 2006 Subject: [Histonet] Histology aide... Message-ID: <61FA27BFA9398E4CA9184FF9B0759B08660D3F@nt_exchange.lmhealth.org> Hello all, I'm trying to get a position established for a Histology aide. We've not had an official aide before and I need to get some ideas about pay scales and job titles. If anyone is willing to share the payscale for an aide and/or their official job title, I would greatly appreciate it. Please respond to private mail below. Thanks. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Hospital Newark, Ohio 43055 mailto:tmcnemar@lmhealth.org From NSEARCY <@t> swmail.sw.org Mon Feb 13 06:52:41 2006 From: NSEARCY <@t> swmail.sw.org (Nita Searcy) Date: Mon Feb 13 06:53:04 2006 Subject: [Histonet] Billing on Send-out Consultations Message-ID: Can someone give me some references regarding the above? (Consultations sent to other pathologists for second opinion/ verification). I know that the "receiving "entity bills codes 88321; 88323; or 88325. How does the sending lab bill patient? Can they? I have a pathologist that insists there must be a mechanism for billing these. I have received conflicting answers. I need a document- reference that verifies whatever is the rule. Thanks From kmerriam2003 <@t> yahoo.com Mon Feb 13 07:12:01 2006 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Mon Feb 13 07:12:10 2006 Subject: [Histonet] Slide scanner for Quantum Dot Fluorescence In-Reply-To: Message-ID: <20060213131201.44774.qmail@web50315.mail.yahoo.com> Hello all, Thanks to everyone for their suggestions, here are the vendors I am looking at: http://www.compucyte.com/icolor.htm http://www.confocal.com/prodsdetail.cfm?ProdID=6 http://www.zeiss.com/4125681F004CA025?Open Hamamatsu also makes a scanner that will have fluorescence available sometime in the summer, here is their website: http://sales.hamamatsu.com/index.php?id=13198981 Kim Carmen Contreras-Sesvold wrote: Can you let me know the results of this query? I am also interested. Thanks Carmen >>> Kim Merriam 02/09/06 7:53 AM >>> Hello All, I am looking for a slide scanner (captures images of entire tissue sections on slides) that has fluorescent capabilities. We currently have an Aperio slide scanner for our routine light microscopy slides, we are very happy with it, but we will be branching out into Quantum Dots and would like a scanner that can do the same thing for fluorescent-stained slides. Does anyone have any ideas or recommendations? Thanks, Kim Kim Merriam Novartis Cambridge, MA --------------------------------- Yahoo! Mail - Helps protect you from nasty viruses. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Kim Merriam Novartis Cambridge, MA --------------------------------- Yahoo! Autos. Looking for a sweet ride? Get pricing, reviews, & more on new and used cars. From cpomajzl <@t> cpllabs.com Mon Feb 13 07:24:51 2006 From: cpomajzl <@t> cpllabs.com (Chris Pomajzl) Date: Mon Feb 13 07:23:44 2006 Subject: [Histonet] Billing on Send-out Consultations References: Message-ID: <002901c630a0$e108d1b0$26fca8c0@CSP> We usually bill the party requesting the second opinion (be that the referring lab, pathologist, physician, patient). >From what I understand, if the consultation is ordered by the physician, then the patient can be billed. But if the pathologist is ordering the consultation, it is the pathologist that is billed. I think that in these cases, the pathologist will sometimes request the physician to order a second opinion so that the patient can be billed. Not sure if this is correct, but that is my understanding. ----- Original Message ----- From: "Nita Searcy" To: Sent: Monday, February 13, 2006 6:52 AM Subject: [Histonet] Billing on Send-out Consultations > Can someone give me some references regarding the above? (Consultations > sent to other pathologists for second opinion/ verification). > > I know that the "receiving "entity bills codes 88321; 88323; or 88325. > > > How does the sending lab bill patient? Can they? I have a pathologist > that insists there must be a mechanism for billing these. I have > received conflicting answers. I need a document- reference that > verifies whatever is the rule. > > Thanks > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sjchtascp <@t> yahoo.com Mon Feb 13 08:16:30 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Mon Feb 13 08:16:39 2006 Subject: [Histonet] Pancreas Controls Desired Message-ID: <20060213141630.30716.qmail@web90209.mail.scd.yahoo.com> Good morning, Does anyone have any spare FFPE pancreas blocks. I work in a research lab and only have acsess to rodent tissue. The tissue, either human or animal, needs to have been fixed in a timly manner so that would eliminate autopsy tissue. Thanks everyone, Steve --------------------------------- Yahoo! Mail Use Photomail to share photos without annoying attachments. From sjchtascp <@t> yahoo.com Mon Feb 13 08:16:25 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Mon Feb 13 08:17:35 2006 Subject: [Histonet] Pancreas Controls Desired Message-ID: <20060213141626.50521.qmail@web90203.mail.scd.yahoo.com> Good morning, Does anyone have any spare FFPE pancreas blocks. I work in a research lab and only have acsess to rodent tissue. The tissue, either human or animal, needs to have been fixed in a timly manner so that would eliminate autopsy tissue. Thanks everyone, Steve --------------------------------- What are the most popular cars? Find out at Yahoo! Autos From Luis.Chiriboga <@t> med.nyu.edu Mon Feb 13 09:38:37 2006 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Mon Feb 13 09:52:14 2006 Subject: [Histonet] Region 1 Symposium Program Available Message-ID: Good Morning Everyone The New York State Histotechnology Society/Region 1 Symposium program is now available on line. Please visit us at www.nyhisto.org The program, registration form and exhibitor registration form are all available for downloading as Adobe Acrobat (PDF) files. We hope to see you there !!!! From jkiernan <@t> uwo.ca Mon Feb 13 10:24:56 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Mon Feb 13 10:24:39 2006 Subject: [Histonet] RE: purple blue haze References: <200602121240.k1CCeFVT032647@outmail.freedom2surf.net> Message-ID: <43F0B2D8.8E0F7B2@uwo.ca> I think Kemlo has to be right here. If the tap water is very "hard" (too much calcium, usually but not always as bicarbonate), it might precipitate calcium sulphate, by metathesis with sulphate ions from the haemalum. Hardness of water due to calcium bicarbonate is called "temporary" because it's lost on heating the water; carbon dioxide is released, solid calcium carbonate deposits in the pipes, kettle etc, and the water becomes "soft," meaning that you don't need to mix much soap with it to wash your feet, socks etc. With hard water you need more soap because some is wasted in forming a useless scum, composed of insoluble calcium salts of higher fatty acids. Water from Lori's hot tap may therefore not contain enough calcium to make a precipitate. Having made this convincing argument, I must confess that I frequently add a splash of limewater (saturated calcium hydroxide) to accelerate blueing of a haemalum nuclear stain, and have never seen a "haze" artifact. However, I do all staining manually, and the diluted limewater follows 30-60 seconds in running (cold, rather hard) tap water - enough to quickly wash away all the surplus haemalum. If I went straight from haemalum into a less generous amount of hard tap water, especially with an added Ca salt, a hazy calcium sulphate precipitate might well happen. I've never done the test. The phrase "run through the stainer" in Lori's report indicates a mechanized H&E (hemalum and eosin) procedure, in which slides may well be exposed to mininimal passage through the steps of the method, to save time and solvents. Rapid slide transfers carry over materials that don't mix well. Bad mixings often make precipitates or emulsions, which are clouds under the microscope. Anyone who follows up this line of enquiry should write it up for publication. There's probably a 3-page paper in this, for 2 days work in the lab. Widely read technique-oriented journals are eager to publish papers that investigate and explain artifacts that occur with routine methods. It helps everyone. John Kiernan Anatomy, UWO London, Canada ----------------------- kemlo wrote: > > Obviously something in the water! If you restain using hot water does the > haze disappear? You must assume I suppose that something crystallises out of > cold water and that the hot water is either 'cleaner' or is sufficiently > warm to dissolve (or keep dissolved) what ever is doing it. > > In London we never blued in tap water, but then you ever did anything with > London water but use it in the toilet. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > lharris@samhealth.org > Sent: 10 February 2006 22:02 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] RE: purple blue haze > > We had a purple blue haze on our H&E's when we used cold tap water run > through the stainer. We had to use hot water and the haze went away. I've > tried to return to using cold water and we get the haze every time. > > Lori A. Harris, HT (ASCP) > Histology Section Leader > GSRMC - Pathology > 3600 NW Samaritan Drive > Corvallis, OR 97330 > 1-541-768-6078 > lharris@samhealth.org From ROrr <@t> enh.org Mon Feb 13 10:50:41 2006 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Mon Feb 13 10:50:51 2006 Subject: [Histonet] Ca-50 ab Message-ID: Vendors! Does anyone have Ca-50 antibody in stock? I've tried several places and it's been discontinued. Let me know!!! Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 From Rcartun <@t> harthosp.org Mon Feb 13 11:22:07 2006 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Mon Feb 13 11:22:38 2006 Subject: [Histonet] CD146 Message-ID: Does anyone have experience doing IHC for CD146 (Mel-CAM) in formalin-fixed tissue? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax From liz <@t> premierlab.com Mon Feb 13 12:03:56 2006 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Mon Feb 13 12:04:13 2006 Subject: [Histonet] PECAM-1 for goat Message-ID: <001a01c630c7$db837b10$95d48a80@Chlipala> Hello everyone What antibody are you using for detecting PECAM-1 in goat? I have the BD antibody for mouse and I also have Dako's CD31 (mouse anti-human) does anyone know if that antibody will work in goat? Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From christiegowan <@t> msn.com Mon Feb 13 12:13:48 2006 From: christiegowan <@t> msn.com (CHRISTIE GOWAN) Date: Mon Feb 13 12:13:56 2006 Subject: [Histonet] Position Opening Message-ID: We have outstanding opportunities for ASCP certified histologists to join our team. Key job responsibilities include the preparation of frozen and paraffin thin sections, the construction of tissue arrays and the performance of research immunohistochemistry assays. These positions will work closely with our Pathology and Biorepository departments in fulfillment of routine and highly customized customer requests. Both experienced histologists and recent grads are encouraged to apply as we have openings at all levels. We offer a highly competitive compensation package which includes 100% company paid health, dental, and life insurances, a 401(k) plan and paid vacation. This is a great opportunity for the right candidate to join a unique new company on the ground floor, and to grow their career as they share in the growth and success of Cytomyx, LLC. Please send resumes to: christie.gowan@cytomyx.com We are conveniently located in historic Lexington, MA. off route 2. We can also be reached at 781-863-9720 ext 260. From mlafrini <@t> csmlab.com Mon Feb 13 12:21:13 2006 From: mlafrini <@t> csmlab.com (Michael LaFriniere) Date: Mon Feb 13 12:22:16 2006 Subject: [Histonet] Re: blue mucin Message-ID: Any help would be appreciated.... I am noticing recently on routine H&E sections, the mucin on colon biopsies only, demonstrating a strong blue color....I know the mucin is picking up the hematoxylin stain and not differentiating in the acid rinse. I was wondering if anybody has demonstrated this recently and the possible corrections to end this aggravation. This leads me to believe that it may be something with the patient prep prior to the biopsy? I am using the Richard Allen (7221) Hematoxylin and routine reagents. Any suggestions would be greatly welcomed! Thank you Michael R. LaFriniere Executive Director Cytology Services of Maryland (CSM) 301-206-2555 ext 27 301-206-2595 fax michael.lafriniere@csmlab.com ----------------------------------------- From mohana_g2002 <@t> yahoo.com Mon Feb 13 12:37:15 2006 From: mohana_g2002 <@t> yahoo.com (MOHANA) Date: Mon Feb 13 12:37:25 2006 Subject: [Histonet] stereology Message-ID: <20060213183715.9136.qmail@web33501.mail.mud.yahoo.com> please can somebody explain me stereological procedure, optical fractionator method and caveliris principle!!!!!!!!!!!!!! mohana Mohana --------------------------------- Yahoo! Mail Use Photomail to share photos without annoying attachments. From lsunhwa <@t> gmail.com Mon Feb 13 12:57:32 2006 From: lsunhwa <@t> gmail.com (Sunhwa Lee) Date: Mon Feb 13 12:57:42 2006 Subject: [Histonet] HELP, Plastic sectioning Message-ID: <177172430602131057s7175ba87j89405acdfac7c69@mail.gmail.com> I am trying to do plastic sectioning with Fish brain and the section is shattered a lot. I've been changing thickness 1-5um, new knife, and embedding materials (JB-4 and Technovit 7100), but not thing works. My slicing conditions are as follows. Any suggestions, please. In advance, thank you so much. 1. Knife : High profile stainless steel knife 2. Microtome: normal rotating Microtome (Leica 820-II) 3. Thickness: 1-5um 4. Cutting angle: 2-3degree 5. Embedding material: GMA (Technovit 7100) 6. Sample: fish brain From gcallis <@t> montana.edu Mon Feb 13 13:25:12 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Feb 13 13:25:22 2006 Subject: [Histonet] PECAM-1 for goat In-Reply-To: <001a01c630c7$db837b10$95d48a80@Chlipala> References: <001a01c630c7$db837b10$95d48a80@Chlipala> Message-ID: <6.0.0.22.1.20060213122330.01b52d68@gemini.msu.montana.edu> Serotec says Clone CO.3E1D4 is supposed to work, not sure what host of the antibody is, got this from their veterinary antibody cross reactivity chart. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From paulah <@t> grandpath.com Mon Feb 13 14:06:11 2006 From: paulah <@t> grandpath.com (Paula Hineline) Date: Mon Feb 13 14:06:20 2006 Subject: [Histonet] stain Message-ID: <000001c630d8$ef93d530$b900a8c0@Pathlab.local> We are seeing some problems with our short-run / small biopsy H&E stained slides. The nuclear detail varies from good to "milky" and indistinct. Nuclear detail can be markedly indistinct with variations occurring from slide to slide and even on the same slide. We are in the process of troubleshooting but I am hoping someone might recognize this problem and have some suggestions. Your responses are much appreciated. Paul Wozniak Laboratory Manager Pathology Laboratory Grandville, Mi 616.530.3344 x 101 From JWEEMS <@t> sjha.org Mon Feb 13 14:13:30 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Mon Feb 13 14:13:39 2006 Subject: [Histonet] Billing on Send-out Consultations Message-ID: <83AACDB0810528418AA106F9AE9B7F7E01305786@sjhaexc02.sjha.org> This has become a real problem in that many consultants will only bill the facility. The best consultants have stopped billing the patient's insurance. We cannot bill professional charges because our pathologists are contract physicians. We go round and round all the time, and usually the hospital ends up absorbing charges. I'm anxious to know how others are handling it. It is my understanding that if a clinician orders a second opinion, the patient's insurance will pay. However, this has to be precertified and all that jazz before this will happen. I have a wonderful lab assistant who works very hard to keep it all together, but we still have to absorb lots of charges. Some of the insurance companies have a hard time understanding that the patient is not going for referral, but that the patient's pathology case is going. It's a very frustrating endeavor that I hope will become more black and white instead of such a gray area. Pathology has so many gray areas in billing. Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Nita Searcy Sent: Monday, February 13, 2006 7:53 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Billing on Send-out Consultations Can someone give me some references regarding the above? (Consultations sent to other pathologists for second opinion/ verification). I know that the "receiving "entity bills codes 88321; 88323; or 88325. How does the sending lab bill patient? Can they? I have a pathologist that insists there must be a mechanism for billing these. I have received conflicting answers. I need a document- reference that verifies whatever is the rule. Thanks _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From KarBieber <@t> aol.com Mon Feb 13 15:09:23 2006 From: KarBieber <@t> aol.com (KarBieber@aol.com) Date: Mon Feb 13 15:09:36 2006 Subject: [Histonet] back-up power Message-ID: <1c5.39f47387.31224f83@aol.com> This is mainly for Mohs people, but I'm hoping others might have good suggestions too. What do you do for back up power for the cryostat in the event of an electrical outage? Thanks, Karen From TJasper <@t> smdc.org Mon Feb 13 15:18:13 2006 From: TJasper <@t> smdc.org (Jasper, Thomas G.) Date: Mon Feb 13 15:18:38 2006 Subject: [Histonet] back-up power Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA07D39141@harrier> Dear Karen, If this is vital I would suggest a dedicated line with UPS. UPS (uninterrupted power supply) is usually a red outlet box, should be available to you through your facilities management people. Perhaps you have other equipment already on UPS? Seems logical to me. Thomas Jasper HT(ASCP)BAS Anatomic Pathology Supervisor SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of KarBieber@aol.com Sent: Monday, February 13, 2006 3:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] back-up power This is mainly for Mohs people, but I'm hoping others might have good suggestions too. What do you do for back up power for the cryostat in the event of an electrical outage? Thanks, Karen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. As required by federal and state laws, you need to hold this information as privileged and confidential. If you have received this communication in error, please notify the sender and destroy all copies of this communication and any attachments. From AnthonyH <@t> chw.edu.au Mon Feb 13 16:07:02 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Mon Feb 13 16:10:01 2006 Subject: [Histonet] Re: blue mucin Message-ID: Michael, The mucin staining is a function of the Hematoxylin pH. Lowering the pH (usually by adding acetic acid) can significantly reduce mucin staining. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michael LaFriniere Sent: Tuesday, 14 February 2006 5:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: blue mucin Any help would be appreciated.... I am noticing recently on routine H&E sections, the mucin on colon biopsies only, demonstrating a strong blue color....I know the mucin is picking up the hematoxylin stain and not differentiating in the acid rinse. I was wondering if anybody has demonstrated this recently and the possible corrections to end this aggravation. This leads me to believe that it may be something with the patient prep prior to the biopsy? I am using the Richard Allen (7221) Hematoxylin and routine reagents. Any suggestions would be greatly welcomed! Thank you Michael R. LaFriniere Executive Director Cytology Services of Maryland (CSM) 301-206-2555 ext 27 301-206-2595 fax michael.lafriniere@csmlab.com ----------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From RSRICHMOND <@t> aol.com Mon Feb 13 17:30:13 2006 From: RSRICHMOND <@t> aol.com (RSRICHMOND@aol.com) Date: Mon Feb 13 17:30:25 2006 Subject: [Histonet] Histobath coolants - hydrofluoroethers Message-ID: <24.1a21423.31227085@aol.com> I asked Histonetters earlier whether they knew of any non-flammable coolant was available for the Thermo Electron Histobath. According to my Thermo Electron Corporation representative, a non-flammable coolant for their Histobath frozen section freezing device is made by 3M. It's 3M? Novec? Engineered Fluid HFE-7100 This product belongs to a class of fluorocarbons called "segregated hydrofluoroethers (HFE's)" According to various MSDS, this HFE-7100 is methyl nonafluoroisobutyl ether C4F9-O-CH3 It melts and freezes at -135 C, so that it will remain liquid in the Histobath. (It would freeze solid in liquid nitrogen, however.) It boils at 60 C., and is listed as non-flammable. It costs around $230 a gallon. Apparently these hydrofluorethers were introduced in 1996 to replace ozone-damaging chlorofluorocarbons, and have since found numerous uses in the laboratory and in industry. What I can't find in Google is any reference to their use for freezing tissue for frozen sections. Is anyone on Histonet familiar with HFE-7100 ? Bob Richmond Gastonia NC From JMacDonald <@t> mtsac.edu Mon Feb 13 17:37:21 2006 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Mon Feb 13 17:37:09 2006 Subject: [Histonet] Fw: Help needed Message-ID: A friend has asked advice using MMA. I have no experience. Can anyone help? See message below. Thank you. Jennifer MacDonald ----- Forwarded by Jennifer MacDonald/Biology/NaturalSciencesDiv/MtSAC on 02/13/2006 03:35 PM ----- "Guerrero, Debbie" 02/13/2006 02:32 PM To cc Subject Help needed I have a question and I'm wondering if you can offer some advice. I'm currently embedding M. cochlea in JB-4 (GMA) and Technovit 9100 New, which is MMA. We are having a very hard times with the sections in MMA sticking to the slides, the completely fall of, even before staining. I've tried what they suggest of putting 30% ETOH to the knife and pick it up on a superfrost slide that has 50% ETOH, but no success. Also tomorrow we are going to try embedd in polyester wax. I've been reading about it and I've found that sections also have difficulty staying on the slide. Any sugestions will be highly appreciated. P.S GMA sections no problems, they have worked fine, even with the H&E. Debbie From kemlo.rogerson <@t> waht.swest.nhs.uk Tue Feb 14 02:35:28 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Tue Feb 14 02:36:02 2006 Subject: [Histonet] Re: blue mucin Message-ID: >From my dim memory I think what is happening is that your haematoxylin is acting as a 'basic' stain (or is it acid?). Normally, as you know, haematin would not stain nucleic acids as it is acidic; the aluminium acts as a mordant as nucleic acids are siderphilic (metal loving). When haematin 'over-oxidises' it forms oxyhaematin and that will stain structures such as mucin and cytoplasm. I've found a new batch of recently prepared or stabilised haematoxylin is usually the answer; have I got me acids and bases the correct way round? Or am I just talking rubbish? Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Michael LaFriniere [mailto:mlafrini@csmlab.com] Sent: Monday, February 13, 2006 6:21 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: blue mucin Any help would be appreciated.... I am noticing recently on routine H&E sections, the mucin on colon biopsies only, demonstrating a strong blue color....I know the mucin is picking up the hematoxylin stain and not differentiating in the acid rinse. I was wondering if anybody has demonstrated this recently and the possible corrections to end this aggravation. This leads me to believe that it may be something with the patient prep prior to the biopsy? I am using the Richard Allen (7221) Hematoxylin and routine reagents. Any suggestions would be greatly welcomed! Thank you Michael R. LaFriniere Executive Director Cytology Services of Maryland (CSM) 301-206-2555 ext 27 301-206-2595 fax michael.lafriniere@csmlab.com ----------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo.rogerson <@t> waht.swest.nhs.uk Tue Feb 14 02:42:02 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Tue Feb 14 02:42:28 2006 Subject: [Histonet] stain Message-ID: 'Milky' nuclear detail could be poor fixation, however it's patchy so that's odd. It could be water in the final clearing agent, it could be contaminated mounting medium (maybe water again), it could be carry over of water from contaminated alcohol to the clearing agent. Used to have this and poor resolution that was finally traced to poor fixation; which oddly can be patchy. Contaminated clearing agent or those fast drying mounting mediums also can cause poor resolution due to the scattering of light. Try taking the coverslip off, rehydrate and dehydrate with fresh alcohols and clearing agent; see if it goes away. If it doesn't then fresh mounting medium and if that fails, post fix the sections and restain. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Paula Hineline [mailto:paulah@grandpath.com] Sent: Monday, February 13, 2006 8:06 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] stain We are seeing some problems with our short-run / small biopsy H&E stained slides. The nuclear detail varies from good to "milky" and indistinct. Nuclear detail can be markedly indistinct with variations occurring from slide to slide and even on the same slide. We are in the process of troubleshooting but I am hoping someone might recognize this problem and have some suggestions. Your responses are much appreciated. Paul Wozniak Laboratory Manager Pathology Laboratory Grandville, Mi 616.530.3344 x 101 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo.rogerson <@t> waht.swest.nhs.uk Tue Feb 14 02:51:07 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Tue Feb 14 02:51:21 2006 Subject: [Histonet] Re: blue mucin Message-ID: Doesn't that just get round the problem of 'over-oxidised' stain? Is that not just curing the symptoms but the problem remains that the haematoxylin is over oxidised? Kemlo Rogerson Weston General Hospital Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: Monday, February 13, 2006 10:07 PM To: Michael LaFriniere; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: blue mucin Michael, The mucin staining is a function of the Hematoxylin pH. Lowering the pH (usually by adding acetic acid) can significantly reduce mucin staining. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michael LaFriniere Sent: Tuesday, 14 February 2006 5:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: blue mucin Any help would be appreciated.... I am noticing recently on routine H&E sections, the mucin on colon biopsies only, demonstrating a strong blue color....I know the mucin is picking up the hematoxylin stain and not differentiating in the acid rinse. I was wondering if anybody has demonstrated this recently and the possible corrections to end this aggravation. This leads me to believe that it may be something with the patient prep prior to the biopsy? I am using the Richard Allen (7221) Hematoxylin and routine reagents. Any suggestions would be greatly welcomed! Thank you Michael R. LaFriniere Executive Director Cytology Services of Maryland (CSM) 301-206-2555 ext 27 301-206-2595 fax michael.lafriniere@csmlab.com ----------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SARAH.REEVES <@t> ekht.nhs.uk Tue Feb 14 05:05:59 2006 From: SARAH.REEVES <@t> ekht.nhs.uk (SARAH REEVES) Date: Tue Feb 14 05:10:22 2006 Subject: [Histonet] Does anyone use p16 with liquid based cytology for gynae and or non-gynae? We are looking to improve Message-ID: Does anyone use p16 with liquid based cytology for gynae and or non-gynae? We are looking to improve methodology as our results are quite variable. Thanks in advance Sarah ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of East Kent Hospitals NHS Trust. If you are not the intended recipient, be advised that you have received this email in error and that any use, dissemination, forwarding, printing or copying of this email is strictly prohibited. If you have received this email in error please notify the system manager at the following email address: root.postmaster@ekht.nhs.uk www.kentandmedway.nhs.uk This footnote also confirms that although this email message has been swept by MIMEsweeper for the presence of computer viruses, it is strongly recommended that you carry out your own virus scan of this message and any attachments. www.mimesweeper.com ********************************************************************** From BMolinari <@t> heart.thi.tmc.edu Tue Feb 14 05:47:09 2006 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Tue Feb 14 05:53:52 2006 Subject: [Histonet] HELP, Plastic sectioning Message-ID: I use the Leica RM 2155 automated microtome and a D profile tungsten carbide knife . My angle is approx. 4 degrees, but I have to adjust that accordingly. I cut at 3um. Be sure everything is tight, tight tight. Good luck. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sunhwa Lee Sent: Monday, February 13, 2006 12:58 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HELP, Plastic sectioning I am trying to do plastic sectioning with Fish brain and the section is shattered a lot. I've been changing thickness 1-5um, new knife, and embedding materials (JB-4 and Technovit 7100), but not thing works. My slicing conditions are as follows. Any suggestions, please. In advance, thank you so much. 1. Knife : High profile stainless steel knife 2. Microtome: normal rotating Microtome (Leica 820-II) 3. Thickness: 1-5um 4. Cutting angle: 2-3degree 5. Embedding material: GMA (Technovit 7100) 6. Sample: fish brain _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cpomajzl <@t> cpllabs.com Tue Feb 14 06:51:37 2006 From: cpomajzl <@t> cpllabs.com (Chris Pomajzl) Date: Tue Feb 14 06:50:32 2006 Subject: [Histonet] stain References: Message-ID: <003401c63165$64b5ece0$26fca8c0@CSP> Could also be inadequate de-paraffinization. ----- Original Message ----- From: "Kemlo Rogerson" To: "'Paula Hineline'" ; Sent: Tuesday, February 14, 2006 2:42 AM Subject: RE: [Histonet] stain > 'Milky' nuclear detail could be poor fixation, however it's patchy so that's > odd. It could be water in the final clearing agent, it could be contaminated > mounting medium (maybe water again), it could be carry over of water from > contaminated alcohol to the clearing agent. > > Used to have this and poor resolution that was finally traced to poor > fixation; which oddly can be patchy. Contaminated clearing agent or those > fast drying mounting mediums also can cause poor resolution due to the > scattering of light. Try taking the coverslip off, rehydrate and dehydrate > with fresh alcohols and clearing agent; see if it goes away. If it doesn't > then fresh mounting medium and if that fails, post fix the sections and > restain. > > Kemlo Rogerson > Pathology Manager > Ext 3311 > DD 01934 647057 > Mob 07749 754194 > > > > > -----Original Message----- > From: Paula Hineline [mailto:paulah@grandpath.com] > Sent: Monday, February 13, 2006 8:06 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] stain > > We are seeing some problems with our short-run / small biopsy H&E > stained slides. The nuclear detail varies from good to "milky" and > indistinct. Nuclear detail can be markedly indistinct with variations > occurring from slide to slide and even on the same slide. We are in the > process of troubleshooting but I am hoping someone might recognize this > problem and have some suggestions. Your responses are much appreciated. > > Paul Wozniak > Laboratory Manager > Pathology Laboratory > Grandville, Mi > 616.530.3344 x 101 > _______________________________________________ > 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 From emerald_lake77 <@t> yahoo.com Tue Feb 14 08:43:19 2006 From: emerald_lake77 <@t> yahoo.com (GT Hebert) Date: Tue Feb 14 08:43:27 2006 Subject: [Histonet] Qdots (quantum dots) mounting media question Message-ID: <20060214144319.59207.qmail@web31710.mail.mud.yahoo.com> Hello, Other than PVA-DABCO as a media for mounting IHC slides using antibodies conjugated with qdots, does anyone have any experience or knowledge of other medias that can be used or that should not be used? I just want to see if I can use anything I already have in house (Vector Lab's mounting medias (various) or Polysciences Aqua-Poly/Mount (Cat# 18606-20). Thank you. Gustave T. Hebert Scientist II Cardiovascular and Metabolic Diseases Wyeth Research Cambridge, MA --------------------------------- What are the most popular cars? Find out at Yahoo! Autos From david.kinsley <@t> spcorp.com Tue Feb 14 08:58:38 2006 From: david.kinsley <@t> spcorp.com (Kinsley, David) Date: Tue Feb 14 08:59:20 2006 Subject: [Histonet] Help with IHC on mouse adipose tissue Message-ID: Hi everyone, An associate of mine is going to be working on samples of mouse adipose tissue. She is looking to perform IHC on paraffin sections. Does anyone have any experience, protocols that they would be willing to share with me? I'm looking for appropriate tissue size, fixation conditions, processing, and antigen retrieval conditions. I'm sorry for the short notice but she is planning to collect tissue samples tomorrow so I don't have much time to research this myself. Any suggestions are greatly appreciated. thanks David Kinsley Schering-Plough Research Institute Kenilworth, NJ 07033 908-740-4669 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From vazquezr <@t> ohsu.edu Tue Feb 14 09:05:49 2006 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Tue Feb 14 09:06:22 2006 Subject: [Histonet] back-up power Message-ID: In the event of a power outage , we have one outlet on the backup generator and run an extension cord. Robyn From vazquezr <@t> ohsu.edu Tue Feb 14 09:06:51 2006 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Tue Feb 14 09:07:26 2006 Subject: [Histonet] back-up power Message-ID: That's what we have. Robyn OHSU From pmarcum <@t> vet.upenn.edu Tue Feb 14 09:37:28 2006 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Tue Feb 14 09:37:46 2006 Subject: [Histonet] HELP, Plastic sectioning In-Reply-To: <177172430602131057s7175ba87j89405acdfac7c69@mail.gmail.com > References: <177172430602131057s7175ba87j89405acdfac7c69@mail.gmail.com> Message-ID: <6.1.1.1.2.20060214101720.01998e90@mail.vet.upenn.edu> At 01:57 PM 2/13/2006, Sunhwa Lee wrote: >I am trying to do plastic sectioning with Fish brain and the section >is shattered a lot. I've been changing thickness 1-5um, new knife, >and embedding materials (JB-4 and Technovit 7100), but not thing >works. My slicing conditions are as follows. Any suggestions, please. > In advance, thank you so much. > >1. Knife : High profile stainless steel knife >2. Microtome: normal rotating Microtome (Leica 820-II) >3. Thickness: 1-5um >4. Cutting angle: 2-3degree >5. Embedding material: GMA (Technovit 7100) >6. Sample: fish brain > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Hi Sunwha, You did not mention how you are fixing and dehydrating the tissues and that can affect sectioning also. What is you schedule and how are you polymerizing? Is it room temperature or 4C? You thickness with the correct knife and tissue preparation is not the problem. As you are using JB4 or Technovit 7100 it will still require a glass knife or tungsten carbide knife at the least for your best sections. That may require a special knife holder for the older blade types as in pre-disposable. I know Linda Jenkins has used disposable knives however, I believe they were the low profiles. Plastics are simply too hard for most disposable knives and the knives will deteriorate too quickly for you get good sections routinely. A standard microtome is usable just not recommended as they are not generally heavy enough to cut smoothly with very hard materials. I have seen people with tons of experience section well on lighter microtomes and I have done some it is just much harder to get consistent sections. Angle is determined by trying different positions and seeing what works best. One angle is not good for all blocks. I change angle sometimes when the block is larger or smaller and it is always while I face the block so I don't lose sections of my tissue. Hope this helps. Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From Inga.Hansson <@t> neuro.uu.se Tue Feb 14 10:05:06 2006 From: Inga.Hansson <@t> neuro.uu.se (Inga Hansson) Date: Tue Feb 14 09:55:20 2006 Subject: [Histonet] methanol treatment Message-ID: Hi all, One of my collegues claims that she sometimes treats paraffin sections with ice-cold methanol in order to get "difficult antibodies" to work. Has anyone of you ever heard of this type of treatment and, if so, can you tell me how it works? many thanks in advance! Inga From jorge.tornero <@t> gmail.com Tue Feb 14 10:06:38 2006 From: jorge.tornero <@t> gmail.com (Jorge Tornero) Date: Tue Feb 14 10:06:49 2006 Subject: [Histonet] HELP, Plastic sectioning In-Reply-To: <8c964a790602140806g612e0ee2p@mail.gmail.com> References: <177172430602131057s7175ba87j89405acdfac7c69@mail.gmail.com> <6.1.1.1.2.20060214101720.01998e90@mail.vet.upenn.edu> <8c964a790602140806g612e0ee2p@mail.gmail.com> Message-ID: <8c964a790602140806vd54f330u@mail.gmail.com> 2006/2/14, Jorge Tornero : > Hi, > > I think the first thing you have to change is your knife. As far as I > know, it must be a tungste-carbide knife (or glass). > > I use a leica rm-2145 microtome whit a D-profile knife. I cut with an > angle of about 6 degrees. > > But also I have suffered from shattering and I was annoyed because we > even use a leica EM-TP tissue proccessor to do the embedding. We use a > very long protocol for dehydration and embedding (24 EtOH 70, 24 h > EtOH 96, 24 h EtOH 100, 48 h 1:1 and 60 hours of pure activated resin) > and it worked for everybody except us. Why? One day I realized that we > put the pure resin and the 1:1 solution at the begginning of the > proccess (when we started the EM-TP) and oviously the resin was > damaged (it turns with a brownish color) when its turn in the > processor began. So now, we put the resin into the processor hours > before its use (because it is friday and we dont work on weekend and > the dwarf inside the processor works for us and changes the liquids) > And now everything is fine. Check if there is a failure in your > protocol or equipment and remember to store any resin or its solutions > in the fridge at 4%. Our problem was caused because the EM-TP only > refrigerates the vial that is being used in that moment. So we trust > that we had 4 ?C in our processor but it only was in the vial that was > running. > > Saludos, > > Jorge Tornero > Instituto Espa?ol de Oceanografia > C?diz - SPAIN > > > 2006/2/14, Pamela Marcum : > > At 01:57 PM 2/13/2006, Sunhwa Lee wrote: > > >I am trying to do plastic sectioning with Fish brain and the section > > >is shattered a lot. I've been changing thickness 1-5um, new knife, > > >and embedding materials (JB-4 and Technovit 7100), but not thing > > >works. My slicing conditions are as follows. Any suggestions, please. > > > In advance, thank you so much. > > > > > >1. Knife : High profile stainless steel knife > > >2. Microtome: normal rotating Microtome (Leica 820-II) > > >3. Thickness: 1-5um > > >4. Cutting angle: 2-3degree > > >5. Embedding material: GMA (Technovit 7100) > > >6. Sample: fish brain > > > > > >_______________________________________________ > > >Histonet mailing list > > >Histonet@lists.utsouthwestern.edu > > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > Hi Sunwha, > > > > You did not mention how you are fixing and dehydrating the tissues and that > > can affect sectioning also. What is you schedule and how are you > > polymerizing? Is it room temperature or 4C? You thickness with the > > correct knife and tissue preparation is not the problem. > > > > As you are using JB4 or Technovit 7100 it will still require a glass knife > > or tungsten carbide knife at the least for your best sections. That may > > require a special knife holder for the older blade types as in > > pre-disposable. I know Linda Jenkins has used disposable knives however, I > > believe they were the low profiles. Plastics are simply too hard for most > > disposable knives and the knives will deteriorate too quickly for you get > > good sections routinely. A standard microtome is usable just not > > recommended as they are not generally heavy enough to cut smoothly with > > very hard materials. I have seen people with tons of experience section > > well on lighter microtomes and I have done some it is just much harder to > > get consistent sections. Angle is determined by trying different positions > > and seeing what works best. One angle is not good for all blocks. I > > change angle sometimes when the block is larger or smaller and it is always > > while I face the block so I don't lose sections of my tissue. Hope this helps. > > > > Best Regards, > > > > Pamela A Marcum > > Manager, Histology Special Procedures > > University of Pennsylvania > > School of Veterinary Medicine > > R.S. Reynolds Jr. CORL > > New Bolton Center > > 382 West Street Road > > Kennett Square, PA 19348 > > > > Phone - 610-925-6278 > > Fax - 610-925-8120 > > E-mail - pmarcum@vet.upenn.edu > > > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From jorge.tornero <@t> gmail.com Tue Feb 14 10:07:17 2006 From: jorge.tornero <@t> gmail.com (Jorge Tornero) Date: Tue Feb 14 10:07:29 2006 Subject: [Histonet] HELP, Plastic sectioning In-Reply-To: <8c964a790602140806vd54f330u@mail.gmail.com> References: <177172430602131057s7175ba87j89405acdfac7c69@mail.gmail.com> <6.1.1.1.2.20060214101720.01998e90@mail.vet.upenn.edu> <8c964a790602140806g612e0ee2p@mail.gmail.com> <8c964a790602140806vd54f330u@mail.gmail.com> Message-ID: <8c964a790602140807s3195cbaem@mail.gmail.com> Hi, I think the first thing you have to change is your knife. As far as I know, it must be a tungste-carbide knife (or glass). I use a leica rm-2145 microtome whit a D-profile knife. I cut with an angle of about 6 degrees. But also I have suffered from shattering and I was annoyed because we even use a leica EM-TP tissue proccessor to do the embedding. We use a very long protocol for dehydration and embedding (24 EtOH 70, 24 h EtOH 96, 24 h EtOH 100, 48 h 1:1 and 60 hours of pure activated resin) and it worked for everybody except us. Why? One day I realized that we put the pure resin and the 1:1 solution at the begginning of the proccess (when we started the EM-TP) and oviously the resin was damaged (it turns with a brownish color) when its turn in the processor began. So now, we put the resin into the processor hours before its use (because it is friday and we dont work on weekend and the dwarf inside the processor works for us and changes the liquids) And now everything is fine. Check if there is a failure in your protocol or equipment and remember to store any resin or its solutions in the fridge at 4%. Our problem was caused because the EM-TP only refrigerates the vial that is being used in that moment. So we trust that we had 4 ?C in our processor but it only was in the vial that was running. Saludos, Jorge Tornero Instituto Espa?ol de Oceanografia C?diz - SPAIN From Michele_Marggi <@t> ssmhc.com Tue Feb 14 10:24:48 2006 From: Michele_Marggi <@t> ssmhc.com (Michele_Marggi@ssmhc.com) Date: Tue Feb 14 10:25:22 2006 Subject: [Histonet] AMACR (P504S) Message-ID: I am researching AMACR (P504s) and need some help. Who out there in histo-land is using it? Is this a Research Use Only antibody? Is a disclaimer in the report required? Any input would be appreciated...... Thanks, Michele Marggi Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. From dellav <@t> musc.edu Tue Feb 14 10:47:56 2006 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Tue Feb 14 10:48:26 2006 Subject: [Histonet] Histobath coolants - hydrofluoroethers Message-ID: Bob, thanks for sharing this information. You may be the pioneer though in the application of HFE 7100 for freezing tissues. you may have a journal article once you've had some experience with the product. you'll want to explore whether HFE 7100 will react in any undesirable fashion with OCT or other embedding media used for frozen sections or with fresh tissues for that matter. any significant safety cautions in the MSDS we should know about? Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> 02/13/06 06:30PM >>> I asked Histonetters earlier whether they knew of any non-flammable coolant was available for the Thermo Electron Histobath. According to my Thermo Electron Corporation representative, a non-flammable coolant for their Histobath frozen section freezing device is made by 3M. It's 3M* Novec* Engineered Fluid HFE-7100 This product belongs to a class of fluorocarbons called "segregated hydrofluoroethers (HFE's)" According to various MSDS, this HFE-7100 is methyl nonafluoroisobutyl ether C4F9-O-CH3 It melts and freezes at -135 C, so that it will remain liquid in the Histobath. (It would freeze solid in liquid nitrogen, however.) It boils at 60 C., and is listed as non-flammable. It costs around $230 a gallon. Apparently these hydrofluorethers were introduced in 1996 to replace ozone-damaging chlorofluorocarbons, and have since found numerous uses in the laboratory and in industry. What I can't find in Google is any reference to their use for freezing tissue for frozen sections. Is anyone on Histonet familiar with HFE-7100 ? Bob Richmond Gastonia NC _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From David.Edmondson <@t> christie-tr.nwest.nhs.uk Tue Feb 14 11:04:10 2006 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Tue Feb 14 11:07:02 2006 Subject: [Histonet] small dryer Message-ID: <3C83687E8F6AE04792E361ABE2D385B84180AA@cht-mail2-2k.xchristie.nhs.uk> SHANDON do one, that is apparently made by Raymond Lamb in the UK Model No. 3120064 We have only had it for a couple of years so the information will not be too much out of date. David -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Maria Christensen Sent: 06 February 2006 21:02 To: Steven Coakley; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] small dryer >Does anyone know where I might be able to get a small forced air dryer, >holding 2-3 racks of slides? > > Steve > > >--------------------------------- >Brings words and photos together (easily) with > PhotoMail - it's free and works with Yahoo! Mail. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet We have one from TBS, Slide Dryer II. It works pretty well for what we use it (dry 130-160 slides O.N. at 37C) but it can dry slides in about half an hour. I think it lists for $1500 at Fisher Scientific, but we get a nice discount from them. Hope this helps! -- Mar?a Maria A. Christensen Technical Associate Dept. of Medical Microbiology & Immunology Creighton University School of Medicine 2500 California Plaza Omaha, Nebraska 68178-0404 voice (402) 280-2678 e-mail mariac@creighton.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ****************************************************************************************************************** This e-mail and any files transmitted with it are confidential and solely for the use of the intended recipient. If you have received this e-mail in error you should not disseminate, distribute or copy it. Please notify the sender immediately and delete this e-mail from your system. ******************************************************************************************************************* From nair.ashwin <@t> gmail.com Tue Feb 14 11:59:55 2006 From: nair.ashwin <@t> gmail.com (Ashwin Nair) Date: Tue Feb 14 12:00:04 2006 Subject: [Histonet] Frigocut 2800 E Message-ID: Hi, We have a Reichert-Jung Frigocut 2800 E cryostat in the lab which we bought recently. However, we didn't get a foot switch or a foot pedal along with it. I was wondering if someone could tell me where I could get one and whether there is anything specific that I need to be aware of when I buy one. Leica has a foot pedal but they are not sure if it will fit in to the 8 amp socket. Could someone please help me. -- Thank you. Ashwin Nair From RSRICHMOND <@t> aol.com Tue Feb 14 12:43:58 2006 From: RSRICHMOND <@t> aol.com (RSRICHMOND@aol.com) Date: Tue Feb 14 12:44:15 2006 Subject: [Histonet] AMACR (P504S) Message-ID: <207.12778982.31237eee@aol.com> Michele Marggi, Surgical Pathology Supervisor, St. Marys Hospital Medical Center, Madison, Wisconsin asks: >>I am researching AMACR (P504s) and need some help. Who out there in histo-land is using it? Is this a Research Use Only antibody? Is a disclaimer in the report required?<< P504S = AMACR, alpha-methylacyl-CoA racemase. An immunostain for AMACR is supposed to mark prostate cancer and not benign glands - also marks many urothelial cancers. Jonathan Epstein, the prostate guru at Johns Hopkins, recommends it. He notes that while all prostate cancers processed at Johns Hopkins mark with it, that 30% of prostate cancers processed elsewhere don't. Clearly some processing problems need to be researched. Our group of five pathologists introduced it a year or so ago. We haven't been too impressed with it (though we find the 34BE12 IHC for prostate basal cell high molecular weight cytokeratin to be essential in maybe 10% of cases). I'm not aware of any special disclaimer needed in a report. Like any "special stain for cancer" it's dangerous in the hands of anybody but an experienced pathologist! Bob Richmond Gastonia NC From NHeath <@t> Lifespan.org Tue Feb 14 12:45:22 2006 From: NHeath <@t> Lifespan.org (Heath, Nancy L.) Date: Tue Feb 14 12:45:38 2006 Subject: [Histonet] plexiglass container Message-ID: <09C945920A6B654199F7A58A1D7D1FDE65ED74@lsexch.lsmaster.lifespan.org> Hi, Does anyone know where I can get a plexiglass container to go around my metler scale?? Thanks, Nancy Heath, HT(ASCP) Neuropathology Dept Rhode Island Hospital nheath@lifespan.org From ROrr <@t> enh.org Tue Feb 14 12:46:59 2006 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Tue Feb 14 12:47:08 2006 Subject: [Histonet] AMACR Message-ID: Michele, I use P504s (AKA) AMACR. I have run both the predilute and the concentrate from Biocare. It works very well in my lab. I also use Biocare's PIN-4 wich is a cocktail of the P504s, p63 and a HMWck The P504s is red. The other 2 are brown (nuclear) and Brown (cytoplasmic) The advantage of the "triple stain" is especially evident when you have one slide of a needle bx of the prostate. You can have a look at all 3 of these stains on one slide. I'll be glad to answer any questions you have about this privately, let me know. As for the designation the data sheet I have from Biocare lists P504s as IVD. I believe this is an updated data sheet. We don't list a specific disclaimer for this antibody on our reports. I'm wondering why you would ask, in case I'm missing something! Hello to all my friends in Madison! Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 ---------------------------------------------------------------------- Message: 1 Date: Tue, 14 Feb 2006 10:24:48 -0600 From: Michele_Marggi@ssmhc.com Subject: [Histonet] AMACR (P504S) To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" I am researching AMACR (P504s) and need some help. Who out there in histo-land is using it? Is this a Research Use Only antibody? Is a disclaimer in the report required? Any input would be appreciated...... Thanks, Michele Marggi Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. From POWELL_SA <@t> Mercer.edu Tue Feb 14 13:21:00 2006 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Tue Feb 14 13:21:24 2006 Subject: [Histonet] plexiglass container In-Reply-To: <09C945920A6B654199F7A58A1D7D1FDE65ED74@lsexch.lsmaster.lifespan.org> Message-ID: <01LYYMKHT1928WWU2I@Macon2.Mercer.edu> A researcher here made me one for my balance. I gave the dimensions and he cut and glued it together. Shirley -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heath, Nancy L. Sent: Tuesday, February 14, 2006 1:45 PM To: Histonet Server (E-mail) Subject: [Histonet] plexiglass container Hi, Does anyone know where I can get a plexiglass container to go around my metler scale?? Thanks, Nancy Heath, HT(ASCP) Neuropathology Dept Rhode Island Hospital nheath@lifespan.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bakarw <@t> hotmail.com Tue Feb 14 13:39:27 2006 From: bakarw <@t> hotmail.com (Barbara Karwinski) Date: Tue Feb 14 13:39:36 2006 Subject: [Histonet] Leica ST4040 Message-ID: Hi, Is somebody using Leica ST4040 autostainer? I need help with staining procedure for HE and Pap. Barbara From ROrr <@t> enh.org Tue Feb 14 13:50:10 2006 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Tue Feb 14 13:50:19 2006 Subject: [Histonet] CORRECTION ON AMACR Message-ID: Hi everyone, I need to make a correction I reviewed my data sheet and checked with Biocare, I found out today, This antibody as it is sold from Biocare is now RUO (research use only). Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 ---------------------------------------------------------------------- Message: 1 Date: Tue, 14 Feb 2006 10:24:48 -0600 From: Michele_Marggi@ssmhc.com Subject: [Histonet] AMACR (P504S) To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" I am researching AMACR (P504s) and need some help. Who out there in histo-land is using it? Is this a Research Use Only antibody? Is a disclaimer in the report required? Any input would be appreciated...... Thanks, Michele Marggi Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 > From david.kinsley <@t> spcorp.com Tue Feb 14 13:59:55 2006 From: david.kinsley <@t> spcorp.com (Kinsley, David) Date: Tue Feb 14 14:00:39 2006 Subject: [Histonet] Suggestions for sectioning mouse colon Message-ID: Hi Everyone, I've been sectioning longitudinal sections of mouse colon and can't seem to alleviate the "pinches" in the colon after staining. Here is what I've been doing; There are 3 longitudinal strips of colon in 1 paraffin block (proximal colon to rectum) Section at 5um and float on water bath 48-50 degrees C Allow sections to float for 1-2 minutes and pick up on + slides Allow sections to dry vertically briefly in a slide rack (to allow excess water to drain off) Lay sections flat on a heating plate at 48-50 degrees C for 1hour Then I place them in a staining rack and heat them in an oven at 60 degrees C for 2 hours to overnight. Stain with H&E I can't seem to be able to obtain nice flat sections all of the time....they don't really have folds, they come off the knife well and look nice and flat prior to staining, but there are many small "pinches" alone the length of the colon. Any suggestions are appreciated. David Kinsley Schering-Plough Research Institute Kenilworth, NJ 07033 908-740-4669 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From AnthonyH <@t> chw.edu.au Tue Feb 14 15:40:36 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Tue Feb 14 15:40:59 2006 Subject: [Histonet] Re: blue mucin Message-ID: Hi Kemlo, As with most problems, there are often several factors that enter into the equation. Old over-oxidised haematoxylins, in my experience tend to colour tissue elements brown but these solutions are definitely old. Fresh haematoxylins, and those that are well within their shelf life tend to stain mucins when they are not acidified enough. A haematoxylin that is a little old and more neutral would probably stain mucins as you have suggested without showing the brown effect. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Kemlo Rogerson [mailto:kemlo.rogerson@waht.swest.nhs.uk] Sent: Tuesday, 14 February 2006 7:51 PM To: Tony Henwood; Michael LaFriniere; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: blue mucin Doesn't that just get round the problem of 'over-oxidised' stain? Is that not just curing the symptoms but the problem remains that the haematoxylin is over oxidised? Kemlo Rogerson Weston General Hospital Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: Monday, February 13, 2006 10:07 PM To: Michael LaFriniere; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: blue mucin Michael, The mucin staining is a function of the Hematoxylin pH. Lowering the pH (usually by adding acetic acid) can significantly reduce mucin staining. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michael LaFriniere Sent: Tuesday, 14 February 2006 5:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: blue mucin Any help would be appreciated.... I am noticing recently on routine H&E sections, the mucin on colon biopsies only, demonstrating a strong blue color....I know the mucin is picking up the hematoxylin stain and not differentiating in the acid rinse. I was wondering if anybody has demonstrated this recently and the possible corrections to end this aggravation. This leads me to believe that it may be something with the patient prep prior to the biopsy? I am using the Richard Allen (7221) Hematoxylin and routine reagents. Any suggestions would be greatly welcomed! Thank you Michael R. LaFriniere Executive Director Cytology Services of Maryland (CSM) 301-206-2555 ext 27 301-206-2595 fax michael.lafriniere@csmlab.com ----------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Sandra.Etheridge <@t> gov.bc.ca Tue Feb 14 16:11:52 2006 From: Sandra.Etheridge <@t> gov.bc.ca (Etheridge, Sandra AL:EX) Date: Tue Feb 14 16:12:02 2006 Subject: [Histonet] Alcohol Based EDTA Decalcifiers Message-ID: <2437188E40DE2947BD3AE43C87F3CB5F12A2A1@stand.idir.bcgov> Hello, everyone, Is anyone aware of a commercial or homemade alcohol based EDTA decalcifying solution? Our fish pathologist is currently fixing his tissues in Davidson's and would prefer to go into an alcohol decal solution instead of an aqueous one (acid hematin pigment forms around the red cells - not nice for photos). Any information is, as always, greatly appreciated. Thanks! Sandra Etheridge BC Ministry of Agriculture & Lands Animal Health Center, Histology Abbotsford, BC Canada From Linresearch <@t> aol.com Tue Feb 14 16:59:39 2006 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Tue Feb 14 16:59:58 2006 Subject: [Histonet] Microarray Message-ID: <286.590e3d4.3123badb@aol.com> Hi, Are there any hints to share on getting all the cores to stay evenly in the recipient block and to get sections without losing cores? Lin From mhwac <@t> yahoo.com Wed Feb 15 02:21:09 2006 From: mhwac <@t> yahoo.com (mh how) Date: Wed Feb 15 02:21:27 2006 Subject: [Histonet] Hacker HCM 6000 coverslipper Message-ID: <20060215082109.54870.qmail@web53003.mail.yahoo.com> Hi ! We are considering getting a HCM 6000 coverslipper from Hacker. Does anybody out there has any experience with this model ? Any comments would be appreciated. Thanks ! Shannon Bita Lifescience Sdn Bhd Kuala Lumpur Malaysia --------------------------------- Yahoo! Mail Use Photomail to share photos without annoying attachments. From Jacqueline.Malam <@t> rli.mbht.nhs.uk Wed Feb 15 02:41:08 2006 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Wed Feb 15 02:46:08 2006 Subject: [Histonet] AMACR Message-ID: Hi We use it regularly in conjunction with 34BE12 on prostatic core biopsies Jacqui Lancaster UK DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From kemlo.rogerson <@t> waht.swest.nhs.uk Wed Feb 15 02:47:20 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Wed Feb 15 02:47:36 2006 Subject: [Histonet] Re: blue mucin Message-ID: I assume that is the function of glycerol in haematoxylin solutions, to stabilise them against overoxidation. It's a memory from those days I made haematoxylin solutions the nice smell of Ehrlich's (was it Ehrlich's?) you got a smell (?an ester) when it was ripe. Ripening big flasks of haematoxylin in the window sill, mine were 'organic' never having the oxidiser added but naturally ripened. Is that when you are truly old? Reminiscing over haematoxylin production? Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: Tuesday, February 14, 2006 9:41 PM To: Kemlo Rogerson; Michael LaFriniere; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: blue mucin Hi Kemlo, As with most problems, there are often several factors that enter into the equation. Old over-oxidised haematoxylins, in my experience tend to colour tissue elements brown but these solutions are definitely old. Fresh haematoxylins, and those that are well within their shelf life tend to stain mucins when they are not acidified enough. A haematoxylin that is a little old and more neutral would probably stain mucins as you have suggested without showing the brown effect. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Kemlo Rogerson [mailto:kemlo.rogerson@waht.swest.nhs.uk] Sent: Tuesday, 14 February 2006 7:51 PM To: Tony Henwood; Michael LaFriniere; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: blue mucin Doesn't that just get round the problem of 'over-oxidised' stain? Is that not just curing the symptoms but the problem remains that the haematoxylin is over oxidised? Kemlo Rogerson Weston General Hospital Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: Monday, February 13, 2006 10:07 PM To: Michael LaFriniere; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: blue mucin Michael, The mucin staining is a function of the Hematoxylin pH. Lowering the pH (usually by adding acetic acid) can significantly reduce mucin staining. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michael LaFriniere Sent: Tuesday, 14 February 2006 5:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: blue mucin Any help would be appreciated.... I am noticing recently on routine H&E sections, the mucin on colon biopsies only, demonstrating a strong blue color....I know the mucin is picking up the hematoxylin stain and not differentiating in the acid rinse. I was wondering if anybody has demonstrated this recently and the possible corrections to end this aggravation. This leads me to believe that it may be something with the patient prep prior to the biopsy? I am using the Richard Allen (7221) Hematoxylin and routine reagents. Any suggestions would be greatly welcomed! Thank you Michael R. LaFriniere Executive Director Cytology Services of Maryland (CSM) 301-206-2555 ext 27 301-206-2595 fax michael.lafriniere@csmlab.com ----------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo.rogerson <@t> waht.swest.nhs.uk Wed Feb 15 03:09:04 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Wed Feb 15 03:09:19 2006 Subject: [Histonet] Alcohol Based EDTA Decalcifiers Message-ID: I don't know if this makes sense but alcohol is non-polar liquid isn't it? Will EDTA work in a non-polar environment? I think the alcohol will suppress ionisation and hence not work; calcium ions are also insoluble in alcohol. 10% solution of EDTA in 0.1M Tris buffer PH 6.95 may be the solution (get it?). Best wishes Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Etheridge, Sandra AL:EX [mailto:Sandra.Etheridge@gov.bc.ca] Sent: Tuesday, February 14, 2006 10:12 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Alcohol Based EDTA Decalcifiers Hello, everyone, Is anyone aware of a commercial or homemade alcohol based EDTA decalcifying solution? Our fish pathologist is currently fixing his tissues in Davidson's and would prefer to go into an alcohol decal solution instead of an aqueous one (acid hematin pigment forms around the red cells - not nice for photos). Any information is, as always, greatly appreciated. Thanks! Sandra Etheridge BC Ministry of Agriculture & Lands Animal Health Center, Histology Abbotsford, BC Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Wed Feb 15 04:21:25 2006 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Wed Feb 15 04:21:55 2006 Subject: [Histonet] Modified SDH Message-ID: <004701c63219$93d88690$2832d445@HPPav2> Our neuropathologist wants to try a procedure he found mentioned in one sentence, in one textbook, no reference, called "modified SDH". It stains JUST the mitochondria in muscle fibers with "red ragged fiber" syndrome. One of my students is taking this on as their project. We already do a SDH (succinic dehydrogenase) and the modified Gomori trichrome, both of which stain normal mitochondria and those with red ragged fibers. This modified SDH is supposed to stain only the red ragged fiber mitochondria. So far, we have found a few articles that mention it, and that it uses either phenazine methosulfate or phenazine methalsufate. However, these articles were written by PhD's or MD's, and again don't detail HOW to do the stain, nor do they list a reference for the procedure. We've been looking for 3 days now. So we're turning to the wonderful people on the Histonet, knowing there is always somone out there who does know something about every subject, and who is willing to share their written procedure with another histotech. Thanks. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 From lpwenk <@t> sbcglobal.net Wed Feb 15 04:26:14 2006 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Wed Feb 15 04:26:44 2006 Subject: [Histonet] Re: blue mucin In-Reply-To: Message-ID: <004801c6321a$401cde10$2832d445@HPPav2> Check the pH. We had this problem before (different vendor). New hematoxylin out of the bottle had a pH about 2.7, and with water carry would quickly get up to 3.0. Add acetic acid, until pH is 2.4-2.5 Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michael LaFriniere Sent: Monday, February 13, 2006 1:21 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: blue mucin Any help would be appreciated.... I am noticing recently on routine H&E sections, the mucin on colon biopsies only, demonstrating a strong blue color....I know the mucin is picking up the hematoxylin stain and not differentiating in the acid rinse. I was wondering if anybody has demonstrated this recently and the possible corrections to end this aggravation. This leads me to believe that it may be something with the patient prep prior to the biopsy? I am using the Richard Allen (7221) Hematoxylin and routine reagents. Any suggestions would be greatly welcomed! Thank you Michael R. LaFriniere Executive Director Cytology Services of Maryland (CSM) 301-206-2555 ext 27 301-206-2595 fax michael.lafriniere@csmlab.com ----------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From godsgirlnow <@t> msn.com Wed Feb 15 08:35:22 2006 From: godsgirlnow <@t> msn.com (Roxanne Soto) Date: Wed Feb 15 08:35:32 2006 Subject: [Histonet] FISH ON URINE CYTOLOGY Message-ID: Hello everyone, Our lab is thinking about starting FISH on urine cytology and I don't exactly know a lot about it. Does anyone have any info on it, like what kind of probe would one use for abnormal urines (kappa, lambda, etc)? Thanks in advance! Roxanne From godsgirlnow <@t> msn.com Wed Feb 15 08:52:41 2006 From: godsgirlnow <@t> msn.com (Roxanne Soto) Date: Wed Feb 15 08:52:51 2006 Subject: [Histonet] FISH ON URINE CYTOLOGY In-Reply-To: Message-ID: Also, does anyone know what the reimbursement is and what the cost is to perform..... thanks again Roxanne ______________________________________________________________ From: "Roxanne Soto" To: HISTONET@PATHOLOGY.SWMED.EDU Subject: [Histonet] FISH ON URINE CYTOLOGY Date: Wed, 15 Feb 2006 09:35:22 -0500 > > Hello everyone, > > Our lab is thinking about starting FISH on urine cytology and I >don't > exactly know a lot about it. Does anyone have any info on it, >like > what kind of probe would one use for abnormal urines (kappa, >lambda, > etc)? > Thanks in advance! > Roxanne >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cpomajzl <@t> cpllabs.com Wed Feb 15 09:17:45 2006 From: cpomajzl <@t> cpllabs.com (Chris Pomajzl) Date: Wed Feb 15 09:15:46 2006 Subject: [Histonet] FISH ON URINE CYTOLOGY References: Message-ID: <006e01c63242$f9b38540$26fca8c0@CSP> Info on Urovysion Fish Kit http://www.labcorp.com/datasets/labcorp/html/chapter/mono/tg005500.htm ----- Original Message ----- From: "Roxanne Soto" To: Sent: Wednesday, February 15, 2006 8:35 AM Subject: [Histonet] FISH ON URINE CYTOLOGY > > Hello everyone, > > Our lab is thinking about starting FISH on urine cytology and I don't > exactly know a lot about it. Does anyone have any info on it, like > what kind of probe would one use for abnormal urines (kappa, lambda, > etc)? > Thanks in advance! > Roxanne > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nair.ashwin <@t> gmail.com Wed Feb 15 12:04:10 2006 From: nair.ashwin <@t> gmail.com (Ashwin Nair) Date: Wed Feb 15 12:05:06 2006 Subject: [Histonet] PLGA frozen section problem Message-ID: Hi, I am trying to section Poly (lactide-co-glycolide){PLGA} scaffolds and subsequently stain them. I am unable to cut wax sections. I tried embedding the scaffold in OCT by placing the scaffold in a mould with OCT for 1 hour under vacuum 20psi. Then the mould was placed in the freezer. 5 & 10 micron sections were cut. However, the sections that I managed to cut using the cryostat tend to be folded and also disintegrate while staining. Has anyone encountered this. I don't have this problem with PLLA scaffolds though as wax sections work fine. Please help. Ashwin From jhaviland <@t> mdanderson.org Wed Feb 15 12:05:58 2006 From: jhaviland <@t> mdanderson.org (jhaviland@mdanderson.org) Date: Wed Feb 15 12:06:07 2006 Subject: [Histonet] membranous staining of live cells Message-ID: Hello Histonetters: I have a friend that wants to stain the cellular membrane of her blastocytes. It can't be fluorescent or hurt the nuclear DNA or use alcohol as a component of the dye. Is there any thing that does this out there? Joie Haviland, HT (ASCP) Department of Pathology Research MD Anderson Cancer Research Center From TJJ <@t> Stowers-Institute.org Wed Feb 15 12:59:49 2006 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Wed Feb 15 13:00:11 2006 Subject: [Histonet] Re: Suggestions for sectioning mouse colon Message-ID: David, it sounds as though you are doing everything right! You might try adding 1 drop of Photoflo to your flotation bath (or some alcohol) to break the surface tension just a little. It could be your paraffin will only relax to a certain extent, too and nothing will help short of trying a different paraffin. I don't know if a few ripples/nips are worth that headache. Good luck! Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From ewrona <@t> yahoo.com Wed Feb 15 13:21:28 2006 From: ewrona <@t> yahoo.com (Erin Wrona) Date: Wed Feb 15 13:21:38 2006 Subject: [Histonet] Jobs in SF bay area? Message-ID: <20060215192128.94111.qmail@web52512.mail.yahoo.com> Hi all, I'm in the market for a new job in the SF bay area - time for a change! Does anyone know of any openings? Thanks, Erin Wrona San Francisco From runyanc <@t> mail.nih.gov Wed Feb 15 13:29:30 2006 From: runyanc <@t> mail.nih.gov (Runyan, Caroline (NIH/NIMH) [F]) Date: Wed Feb 15 13:29:39 2006 Subject: [Histonet] heat resistant staining nets Message-ID: Hello, I normally perform DAB staining on free-floating brain sections in staining nets from brain research labs. However, I need to add an antigen retrieval step at the beginning, 15 min in citrate buffer at 90C. I've just tested the nets that I have, and they will not stand up to the heat. Does anyone know of a heat-resistant staining net? Thank you. Caroline From gpbnas <@t> yahoo.es Wed Feb 15 13:59:22 2006 From: gpbnas <@t> yahoo.es (Guillermo Palao) Date: Wed Feb 15 13:59:34 2006 Subject: [Histonet] DAPI staining for apoptotic nuclei in frozen sections Message-ID: <20060215195922.71419.qmail@web26210.mail.ukl.yahoo.com> Hello all, I am trying to detect apoptotic nuclei by DAPI staining as a morphological confirmation of FITC-TUNEL positive cells in frozen sections. I stain with DAPI 0.5 ug/ml in PBS for 5 min RT, wash in PBS once, mounting medium and cover. The problem is my DAPI signal is very strong, all nuclei look almost white instead of blue, also nucleoli do not look like "dark spots" in nuclei as in published images, but as very bright dense small white dots. Is it just a question of DAPI concentration? If so, how much could I decrease it? Most articles employ 1 ug/ml for apoptosis detection so I am concerned I might be doing something wrong. Any help is really appreciated. Guillermo Palao, MD. Ph.D. Laboratorio de Reumatolog?a Centro de Investigaci?n Hospital 12 de Octubre Avda de C?rdoba s/n Madrid 28041 Spain --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com From sjchtascp <@t> yahoo.com Wed Feb 15 14:23:02 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Wed Feb 15 14:23:14 2006 Subject: [Histonet] (no subject) Message-ID: <20060215202302.12615.qmail@web90203.mail.scd.yahoo.com> wheres my post of 2/15? --------------------------------- Yahoo! Mail Use Photomail to share photos without annoying attachments. From JWEEMS <@t> sjha.org Wed Feb 15 14:25:18 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Wed Feb 15 14:25:28 2006 Subject: [Histonet] (no subject) Message-ID: <83AACDB0810528418AA106F9AE9B7F7E013057D9@sjhaexc02.sjha.org> here Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Steven Coakley Sent: Wednesday, February 15, 2006 3:23 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) wheres my post of 2/15? --------------------------------- Yahoo! Mail Use Photomail to share photos without annoying attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From awatanabe <@t> tgen.org Wed Feb 15 14:48:52 2006 From: awatanabe <@t> tgen.org (Aprill Watanabe) Date: Wed Feb 15 14:49:01 2006 Subject: [Histonet] Re: Microarray In-Reply-To: <20060215184512.E0EEA2006130@mr1.tgen.org> Message-ID: Lin, I work with TMAs all the time. I'm not sure what your problem is? Are you hand punching the arrays or using an instrument? As far as cores falling off, are they coming off in your applications or when you cut them? Next what tissue are you working with? I have lots of hints but I need you to narrow down the possibilities. If you would prefer you can email me directly or call me for help. On 2/15/06 11:45 AM, "histonet-request@lists.utsouthwestern.edu" wrote: Aprill Watanabe, B.S. Research Associate Tissue Microarray Center (TMA) Translational Genomics Research Institute (TGen) 602-343-8822 awatanabe@tgen.org www.tgen.org From AnthonyH <@t> chw.edu.au Wed Feb 15 16:25:24 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Wed Feb 15 16:27:13 2006 Subject: [Histonet] Re: blue mucin Message-ID: Ha Ha, I remember the story of one lab who also made up Hx the natural way (??) and left it on the window sill to ripen, unfortunately, the lab was on the lower ground floor and it seems the young school boys used to use the opened brown bottle for target practice while they were waiting for the bus. Reminds me of the reaction of orcein and ammonia!! Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Kemlo Rogerson [mailto:kemlo.rogerson@waht.swest.nhs.uk] Sent: Wednesday, 15 February 2006 7:47 PM To: Tony Henwood; Michael LaFriniere; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: blue mucin I assume that is the function of glycerol in haematoxylin solutions, to stabilise them against overoxidation. It's a memory from those days I made haematoxylin solutions the nice smell of Ehrlich's (was it Ehrlich's?) you got a smell (?an ester) when it was ripe. Ripening big flasks of haematoxylin in the window sill, mine were 'organic' never having the oxidiser added but naturally ripened. Is that when you are truly old? Reminiscing over haematoxylin production? Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: Tuesday, February 14, 2006 9:41 PM To: Kemlo Rogerson; Michael LaFriniere; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: blue mucin Hi Kemlo, As with most problems, there are often several factors that enter into the equation. Old over-oxidised haematoxylins, in my experience tend to colour tissue elements brown but these solutions are definitely old. Fresh haematoxylins, and those that are well within their shelf life tend to stain mucins when they are not acidified enough. A haematoxylin that is a little old and more neutral would probably stain mucins as you have suggested without showing the brown effect. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Kemlo Rogerson [mailto:kemlo.rogerson@waht.swest.nhs.uk] Sent: Tuesday, 14 February 2006 7:51 PM To: Tony Henwood; Michael LaFriniere; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: blue mucin Doesn't that just get round the problem of 'over-oxidised' stain? Is that not just curing the symptoms but the problem remains that the haematoxylin is over oxidised? Kemlo Rogerson Weston General Hospital Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: Monday, February 13, 2006 10:07 PM To: Michael LaFriniere; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: blue mucin Michael, The mucin staining is a function of the Hematoxylin pH. Lowering the pH (usually by adding acetic acid) can significantly reduce mucin staining. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michael LaFriniere Sent: Tuesday, 14 February 2006 5:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: blue mucin Any help would be appreciated.... I am noticing recently on routine H&E sections, the mucin on colon biopsies only, demonstrating a strong blue color....I know the mucin is picking up the hematoxylin stain and not differentiating in the acid rinse. I was wondering if anybody has demonstrated this recently and the possible corrections to end this aggravation. This leads me to believe that it may be something with the patient prep prior to the biopsy? I am using the Richard Allen (7221) Hematoxylin and routine reagents. Any suggestions would be greatly welcomed! Thank you Michael R. LaFriniere Executive Director Cytology Services of Maryland (CSM) 301-206-2555 ext 27 301-206-2595 fax michael.lafriniere@csmlab.com ----------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From emry <@t> u.washington.edu Wed Feb 15 19:18:00 2006 From: emry <@t> u.washington.edu (Trisha Emry) Date: Wed Feb 15 19:18:10 2006 Subject: [Histonet] Richard Allen #9 Message-ID: <001401c63296$d41f1d90$8c775f80@sod.washington.edu> A co-worker has just recommended Richard Allen #9 paraffin. Do any of you have some experience with this? I was looking for something to help with wrinkles in large specimens. Thanks, Trisha Seattle From gopher1012 <@t> aol.com Wed Feb 15 21:21:42 2006 From: gopher1012 <@t> aol.com (gopher1012@aol.com) Date: Wed Feb 15 21:21:58 2006 Subject: [Histonet] New List Member Message-ID: <8C800BF3DD00424-C54-C6E@MBLK-M11.sysops.aol.com> Hi, my name is Cassondra Brooks and I am new to Histonet. I decided to join this mailing list because I am currently enrolled in a histology class and I thought this would be a great way to learn more information or clarify present information. I find histology very interesting, especially abnormalites, so any information you would like to pass on would be great! I look foward to learning more information through Histonet. From fadiafandi <@t> mac.com Wed Feb 15 22:27:21 2006 From: fadiafandi <@t> mac.com (fadiafandi@mac.com) Date: Wed Feb 15 22:27:32 2006 Subject: [Histonet] Immediate need for highly experienced Technologists in IHC/ISH/FISH Message-ID: <6E0E281F-0EE6-491D-AB54-C49A7CECAC10@mac.com> This is a posting for an immediate need for multiple positions of highly experienced technologists in Immunohistochemistry, in-situ hybridization (including FISH). Required qualifications include: *Experience for at least seven years in the field working in a high- volume laboratory *Experience with multiple types of IHC instruments and multiple detection systems *Experience with work-up and troubleshooting of instrument failure and antibody staining problems *Experience with validation of >150 antibodies *Experience with validation and work-up of FISH probes (This is not an absolute requirement, but it is desired) *Experience in training and willingness to teach other techs Qualified technologists should submit their curriculum vitae with a short personal statement to Dr. Hadi Yaziji. Please do not email your cv through this email list. Instead, please send an email directly to fadiafandi@mac.com. I will be happy to share additional information with you if you are interested in the position. Best regards, Hadi Yaziji, M.D. From Lchausse <@t> nmh.org Wed Feb 15 22:55:51 2006 From: Lchausse <@t> nmh.org (Chaussey, Leslie) Date: Wed Feb 15 23:00:15 2006 Subject: [Histonet] (no subject) References: <6E0E281F-0EE6-491D-AB54-C49A7CECAC10@mac.com> Message-ID: Hoping someone out there has a better memory than I do. I recall seeing an ad once for a high-density storage microscope slide file system that rotated the appropriate slide cabinet to the user (similar to the clothes coming to the front counter person in a dry cleaner's). Anybody out there recall seeing something like that or have any recollection of the vendor? Thanks for your help! Leslie Chaussey Northwestern Memorial Hospital ----------------------------------------- This message and any included attachments are intended only for the addressee. The information contained in this message is confidential and may constitute proprietary or non-public information under international, federal, or state laws. 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. From kemlo.rogerson <@t> waht.swest.nhs.uk Thu Feb 16 02:34:07 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Thu Feb 16 02:34:25 2006 Subject: [Histonet] Re: blue mucin Message-ID: Would uric acid make it less likely to stain mucin? Downward shots were always easier I remember than upwards! Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: Wednesday, February 15, 2006 10:25 PM To: Kemlo Rogerson; Michael LaFriniere; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: blue mucin Ha Ha, I remember the story of one lab who also made up Hx the natural way (??) and left it on the window sill to ripen, unfortunately, the lab was on the lower ground floor and it seems the young school boys used to use the opened brown bottle for target practice while they were waiting for the bus. Reminds me of the reaction of orcein and ammonia!! Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Kemlo Rogerson [mailto:kemlo.rogerson@waht.swest.nhs.uk] Sent: Wednesday, 15 February 2006 7:47 PM To: Tony Henwood; Michael LaFriniere; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: blue mucin I assume that is the function of glycerol in haematoxylin solutions, to stabilise them against overoxidation. It's a memory from those days I made haematoxylin solutions the nice smell of Ehrlich's (was it Ehrlich's?) you got a smell (?an ester) when it was ripe. Ripening big flasks of haematoxylin in the window sill, mine were 'organic' never having the oxidiser added but naturally ripened. Is that when you are truly old? Reminiscing over haematoxylin production? Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: Tuesday, February 14, 2006 9:41 PM To: Kemlo Rogerson; Michael LaFriniere; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: blue mucin Hi Kemlo, As with most problems, there are often several factors that enter into the equation. Old over-oxidised haematoxylins, in my experience tend to colour tissue elements brown but these solutions are definitely old. Fresh haematoxylins, and those that are well within their shelf life tend to stain mucins when they are not acidified enough. A haematoxylin that is a little old and more neutral would probably stain mucins as you have suggested without showing the brown effect. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Kemlo Rogerson [mailto:kemlo.rogerson@waht.swest.nhs.uk] Sent: Tuesday, 14 February 2006 7:51 PM To: Tony Henwood; Michael LaFriniere; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: blue mucin Doesn't that just get round the problem of 'over-oxidised' stain? Is that not just curing the symptoms but the problem remains that the haematoxylin is over oxidised? Kemlo Rogerson Weston General Hospital Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: Monday, February 13, 2006 10:07 PM To: Michael LaFriniere; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: blue mucin Michael, The mucin staining is a function of the Hematoxylin pH. Lowering the pH (usually by adding acetic acid) can significantly reduce mucin staining. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michael LaFriniere Sent: Tuesday, 14 February 2006 5:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: blue mucin Any help would be appreciated.... I am noticing recently on routine H&E sections, the mucin on colon biopsies only, demonstrating a strong blue color....I know the mucin is picking up the hematoxylin stain and not differentiating in the acid rinse. I was wondering if anybody has demonstrated this recently and the possible corrections to end this aggravation. This leads me to believe that it may be something with the patient prep prior to the biopsy? I am using the Richard Allen (7221) Hematoxylin and routine reagents. Any suggestions would be greatly welcomed! Thank you Michael R. LaFriniere Executive Director Cytology Services of Maryland (CSM) 301-206-2555 ext 27 301-206-2595 fax michael.lafriniere@csmlab.com ----------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nsnwl <@t> neuro.hfh.edu Thu Feb 16 07:24:55 2006 From: nsnwl <@t> neuro.hfh.edu (Nancy Lemke) Date: Thu Feb 16 07:38:47 2006 Subject: [Histonet] whole brain in paraffin Message-ID: <1c215d2a1142f946ddcd4fd83e418828@neuro.hfh.edu> Good Morning Histonet, I would like some information about the preparation and sectioning of 1-2cm slabs of human brain. The cutting will be done on an old Reichert Tetrander (sliding) microtome with a giant steel blade. If anyone who has experience with this sort of sectioning would email me I would love to ask some questions. Thank you in advance, Nancy Lemke Research Coordinator Hermelin Brain Tumor Center Henry Ford Hospital Detroit ============================================================================== Go to http://henryford.com We're Henry Ford. We Can. HFHS CONFIDENTIALITY NOTICE: This email contains information from the sender that may be CONFIDENTIAL, LEGALLY PRIVILEGED, PROPRIETARY or otherwise protected from disclosure. This email is intended for use only by the person or entity to whom it is addressed. If you are not the intended recipient, any use, disclosure, copying, distribution, printing, or any action taken in reliance on the contents of this email, is strictly prohibited. If you received this email in error, please contact the sending party by replying in an email to the sender, delete the email from your computer system and shred any paper copies of the email you printed. Note to Patients: There are a number of risks you should consider before using e-mail to communicate with us. These risks are described in our Privacy Policy at http://henryford.com. Review that policy carefully before continuing to communicate with us by e-mail. For greater Internet security, our policy describes the Henry Ford MyHealth electronic communication process - you may register at http://henryford.com. If you do not believe that our policy gives you the privacy and security protection you need, do not send e-mail or Internet communications to us. ============================================================================== From kemlo.rogerson <@t> waht.swest.nhs.uk Thu Feb 16 08:18:02 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Thu Feb 16 08:18:16 2006 Subject: [Histonet] whole brain in paraffin Message-ID: WOW a Tetrander!!! Thought those things had been consigned to the torture chamber..... Used that thing 25 years ago with mega brain slices, hardest bit was keeping the block cool but as long as the processing was Ok (used to take weeks to process the brain) it was not too bad; needed skill in manipulating the sections and then floating onto dirty great big coated slides. If you have any probs then contact me. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Nancy Lemke [mailto:nsnwl@neuro.hfh.edu] Sent: Thursday, February 16, 2006 1:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] whole brain in paraffin Good Morning Histonet, I would like some information about the preparation and sectioning of 1-2cm slabs of human brain. The cutting will be done on an old Reichert Tetrander (sliding) microtome with a giant steel blade. If anyone who has experience with this sort of sectioning would email me I would love to ask some questions. Thank you in advance, Nancy Lemke Research Coordinator Hermelin Brain Tumor Center Henry Ford Hospital Detroit ============================================================================ == Go to http://henryford.com We're Henry Ford. We Can. HFHS CONFIDENTIALITY NOTICE: This email contains information from the sender that may be CONFIDENTIAL, LEGALLY PRIVILEGED, PROPRIETARY or otherwise protected from disclosure. This email is intended for use only by the person or entity to whom it is addressed. If you are not the intended recipient, any use, disclosure, copying, distribution, printing, or any action taken in reliance on the contents of this email, is strictly prohibited. If you received this email in error, please contact the sending party by replying in an email to the sender, delete the email from your computer system and shred any paper copies of the email you printed. Note to Patients: There are a number of risks you should consider before using e-mail to communicate with us. These risks are described in our Privacy Policy at http://henryford.com. Review that policy carefully before continuing to communicate with us by e-mail. For greater Internet security, our policy describes the Henry Ford MyHealth electronic communication process - you may register at http://henryford.com. If you do not believe that our policy gives you the privacy and security protection you need, do not send e-mail or Internet communications to us. ============================================================================ == _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kmerriam2003 <@t> yahoo.com Thu Feb 16 08:27:53 2006 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Thu Feb 16 08:28:03 2006 Subject: [Histonet] OCT embedding of NBF-fixed specimen Message-ID: <20060216142753.7587.qmail@web50314.mail.yahoo.com> Hello all, I need to do some OCT embedding of tissues that are already fixed in formalin. What is the best way to do this? Should I rinse the tissues in water to get out the formalin? How can I prevent crystal artifact on these tissues? Any advice would be helpful, Thanks, Kim Kim Merriam Novartis Cambridge, MA --------------------------------- Brings words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. From livieira <@t> ualg.pt Thu Feb 16 09:44:52 2006 From: livieira <@t> ualg.pt (Lina Vieira) Date: Thu Feb 16 09:46:03 2006 Subject: [Histonet] Aluminium in paraffin embedded sections Message-ID: <002601c6330f$ed2a3490$2914100a@labhistologia> We want identify Aluminium in paraffin embedded sections of fish spleen and kidney for light microscopy. I found diferent techniques and want your advise about it. I indicate the technique that seems easy to me: (from Pearse, 1972) Solution A . 0.2% solochrome azurine (aqueous) Solution B. 0.2% solochrome azurine in normal sodium hydroxide Method 1. Take two sections to water 2. Stain one section in each solutions - 20 minutes 3. Wash in water 4. Counterstain in 1% neutral red - 5 minutes 5. Wash in tap water 6. Dehydrate through gradde alcohols to xylene 7 Mount in DPX Resultes section stain with solution A - Aluminium and beryllium - blues section stain with solution B - Beryllium - blue-black Thanks in advance for any information you can provide, Lina Vieira University of Algarve Portugal From SAllen <@t> exchange.hsc.mb.ca Thu Feb 16 09:48:28 2006 From: SAllen <@t> exchange.hsc.mb.ca (Sharon Allen) Date: Thu Feb 16 09:49:52 2006 Subject: [Histonet] whole brain in paraffin Message-ID: <36FE435D6D2F1D489174B22362A961B66B8315@hscxntmx0006.hsc.mb.ca> Hi, I have done whole mounts but not for about 6 years (the neuropathologist that used to like them has left - hurray!!). I am including our method for processing the block. I hope this is of some help, please email me if you have any questions. Sharon Allen HSC Winnipeg, MB, CA sallen@hsc.mb.caHAND *Sorry about the uppercase, just the way our manual was typed. PROCESSING WHOLE BRAIN SPECIMENS HAVE SPECIMEN CUT AT A THICKNESS OF 1.5 TO 2.5 CM. ANY THINNER AND THE SPECIMEN MAY CURL, MAKING IT DIFFICULT TO EMBED FLAT. THE PROCESSING TIMES DEPENDS MAINLY ON THE SIZE AND THICKNESS OF THE TISSUE. THE TIMES USED IN THE FOLLOWING LIST WAS FOR A SPECIMEN WHICH WAS 2 - 3 CM IN THICKNESS AND 10 CM IN WIDTH. THE PROCESSING WAS SUCCESSFUL. THE TISSUE CAN BE LEFT IN FORMALIN, WAX, 70% AND 95% ALCOHOL , FOR AN INDEFINITE TIME WITHOUT HARM. *CHANGE THE SOLUTIONS A FEW TIMES PER DAY. FORMALIN-----------------------------------APROX. 1 WEEK 70% ALCOHOL------------------------------OVERNIGHT 70% ALCOHOL------------------------------OVERNIGHT 95% ALCOHOL------------------------------OVERNIGHT 95% ALCOHOL------------------------------OVERNIGHT 95% ALCOHOL------------------------------OVERNIGHT 100% ALCOHOL----------------------------OVERNIGHT 100% ALCOHOL----------------------------OVERNIGHT 100% ALCOHOL----------------------------OVERNIGHT CHANGE TO XYLOL, CHANGING SOLUTION EVERY HOUR AT FIRST. KEEP CHECKING TO SEE IF IT IS CLEARING. (Hold section up to a light) THE TISSUE WILL BECOME MORE TRANSLUCENT AND A LIGHTER COLOR AS IT CLEARS. THE CLEARING WILL TAKE PLACE FROM THE OUTSIDE, INWARD; MAKE SURE THE CENTER IS CLEARED. XYLOL WILL HARDEN THE TISSUE, SO CARE MUST BE TAKEN WITH THE ENDPOINT AND TIMING. WHEN THE CLEARING IS COMPLETE, PUT THE SPECIMEN IN WAX, CHANGING THE SOLUTION OFTEN. THE VACUMM OVEN CAN BE USED AT THIS POINT. SET IT AT THE MELTING POINT OF THE WAX AND AT 15 LBS. PRESSURE FOR 1 HOUR AT A TIME, THEN CHANGE THE WAX, REPEATING THE INFILTRATION UNDER VACUMM. THIS PROCESS CAN TAKE 2 TO 5 DAYS, (INFILTRATION IN THE WAX CAN NOT HARM THE TISSUE) FREQUENTLY CHANGING THE WAX, AND LEAVING THE SPECIMEN IN A 54?C OVEN OVERNIGHT OR FOR THE WEEKEND, IF NECESSARY. THE SPECIMEN IS THEN EMBEDDED AND ATTACHED TO THE CHUCK USED FOR THE SLEDGE MICROTOME. HEAT THE CHUCK, PLACE IT ON THE BACK OF THE BLOCK AND THEN SUBMERGING IT INTO A SINK FILLED WITH ICE COLD WATER. CUT SECTION ON THE SLEDGE MICROTOME AT 6 TO8 MICRONS. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Nancy Lemke Sent: February 16, 2006 7:25 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] whole brain in paraffin Good Morning Histonet, I would like some information about the preparation and sectioning of 1-2cm slabs of human brain. The cutting will be done on an old Reichert Tetrander (sliding) microtome with a giant steel blade. If anyone who has experience with this sort of sectioning would email me I would love to ask some questions. Thank you in advance, Nancy Lemke Research Coordinator Hermelin Brain Tumor Center Henry Ford Hospital Detroit ============================================================================ == Go to http://henryford.com We're Henry Ford. We Can. HFHS CONFIDENTIALITY NOTICE: This email contains information from the sender that may be CONFIDENTIAL, LEGALLY PRIVILEGED, PROPRIETARY or otherwise protected from disclosure. This email is intended for use only by the person or entity to whom it is addressed. If you are not the intended recipient, any use, disclosure, copying, distribution, printing, or any action taken in reliance on the contents of this email, is strictly prohibited. If you received this email in error, please contact the sending party by replying in an email to the sender, delete the email from your computer system and shred any paper copies of the email you printed. Note to Patients: There are a number of risks you should consider before using e-mail to communicate with us. These risks are described in our Privacy Policy at http://henryford.com. Review that policy carefully before continuing to communicate with us by e-mail. For greater Internet security, our policy describes the Henry Ford MyHealth electronic communication process - you may register at http://henryford.com. If you do not believe that our policy gives you the privacy and security protection you need, do not send e-mail or Internet communications to us. ============================================================================ == _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail and/or any documents in this transmission is intended for the address(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. From Terry.Marshall <@t> rothgen.nhs.uk Thu Feb 16 10:03:30 2006 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Thu Feb 16 10:01:52 2006 Subject: [Histonet] whole brain in paraffin/microtome for sale? Message-ID: Talking of tetranders, has anybody in the UK got a decent sledge microtome to sell, preferably for a song. (I can do guitar accompaniment if necessary)? Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:kemlo.rogerson@waht.swest.nhs.uk] Sent: 16 February 2006 14:18 To: 'Nancy Lemke'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] whole brain in paraffin WOW a Tetrander!!! Thought those things had been consigned to the torture chamber..... Used that thing 25 years ago with mega brain slices, hardest bit was keeping the block cool but as long as the processing was Ok (used to take weeks to process the brain) it was not too bad; needed skill in manipulating the sections and then floating onto dirty great big coated slides. If you have any probs then contact me. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Nancy Lemke [mailto:nsnwl@neuro.hfh.edu] Sent: Thursday, February 16, 2006 1:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] whole brain in paraffin Good Morning Histonet, I would like some information about the preparation and sectioning of 1-2cm slabs of human brain. The cutting will be done on an old Reichert Tetrander (sliding) microtome with a giant steel blade. If anyone who has experience with this sort of sectioning would email me I would love to ask some questions. Thank you in advance, Nancy Lemke Research Coordinator Hermelin Brain Tumor Center Henry Ford Hospital Detroit ============================================================================ == Go to http://henryford.com We're Henry Ford. We Can. HFHS CONFIDENTIALITY NOTICE: This email contains information from the sender that may be CONFIDENTIAL, LEGALLY PRIVILEGED, PROPRIETARY or otherwise protected from disclosure. This email is intended for use only by the person or entity to whom it is addressed. If you are not the intended recipient, any use, disclosure, copying, distribution, printing, or any action taken in reliance on the contents of this email, is strictly prohibited. If you received this email in error, please contact the sending party by replying in an email to the sender, delete the email from your computer system and shred any paper copies of the email you printed. Note to Patients: There are a number of risks you should consider before using e-mail to communicate with us. These risks are described in our Privacy Policy at http://henryford.com. Review that policy carefully before continuing to communicate with us by e-mail. For greater Internet security, our policy describes the Henry Ford MyHealth electronic communication process - you may register at http://henryford.com. If you do not believe that our policy gives you the privacy and security protection you need, do not send e-mail or Internet communications to us. ============================================================================ == _______________________________________________ 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 From fadiafandi <@t> mac.com Thu Feb 16 10:04:59 2006 From: fadiafandi <@t> mac.com (fadiafandi@mac.com) Date: Thu Feb 16 10:05:12 2006 Subject: [Histonet] Re: immediate need for experienced techs in IHC/ISH/FISH Message-ID: Dear Histonet members, Regarding the job posting that I shared yesterday (see below), I was informed that I should mention the location of our lab. We're in Miami, Florida. The position doesn't require physical presence in Miami (except for a few people who will be working in our core facility here in Miami). Given the various aspects of our organization, we can either work with you on an employment basis or contractual basis without you having to relocate. I will be happy to share all the details with interested colleagues who can email me directly at fadiafandi@mac.com. Please attach an updated curriculum vitae to your email. Thank you. Hadi Yaziji, M.D. President, Ancillary Pathways, LLC. Miami, FL This is a posting for an immediate need for multiple positions of highly experienced technologists in Immunohistochemistry, in-situ hybridization (including FISH). Required qualifications include: *Experience for at least seven years in the field working in a high- volume laboratory *Experience with multiple types of IHC instruments and multiple detection systems *Experience with work-up and troubleshooting of instrument failure and antibody staining problems *Experience with validation of >150 antibodies *Experience with validation and work-up of FISH probes (This is not an absolute requirement, but it is desired) *Experience in training and willingness to teach other techs Qualified technologists should submit their curriculum vitae with a short personal statement to Dr. Hadi Yaziji. Please do not email your cv through this email list. Instead, please send an email directly to fadiafandi@mac.com. I will be happy to share additional information with you if you are interested in the position. Best regards, Hadi Yaziji, M.D. From kemlo.rogerson <@t> waht.swest.nhs.uk Thu Feb 16 10:27:00 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Thu Feb 16 10:27:17 2006 Subject: [Histonet] Aluminium in paraffin embedded sections Message-ID: The aurine technic of PC Irwin was recommended by Lillie; page 534 of his book Hitopathologic Technic and Practical Histochemistry. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Lina Vieira [mailto:livieira@ualg.pt] Sent: Thursday, February 16, 2006 3:45 PM To: Histonet Subject: [Histonet] Aluminium in paraffin embedded sections We want identify Aluminium in paraffin embedded sections of fish spleen and kidney for light microscopy. I found diferent techniques and want your advise about it. I indicate the technique that seems easy to me: (from Pearse, 1972) Solution A . 0.2% solochrome azurine (aqueous) Solution B. 0.2% solochrome azurine in normal sodium hydroxide Method 1. Take two sections to water 2. Stain one section in each solutions - 20 minutes 3. Wash in water 4. Counterstain in 1% neutral red - 5 minutes 5. Wash in tap water 6. Dehydrate through gradde alcohols to xylene 7 Mount in DPX Resultes section stain with solution A - Aluminium and beryllium - blues section stain with solution B - Beryllium - blue-black Thanks in advance for any information you can provide, Lina Vieira University of Algarve Portugal _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo.rogerson <@t> waht.swest.nhs.uk Thu Feb 16 10:30:16 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Thu Feb 16 10:30:32 2006 Subject: [Histonet] whole brain in paraffin/microtome for sale? Message-ID: Dave at Blackburn have you got an old sledge for Terry Marshall? You squirreled everything else there and converted to rotaries; interestingly they use sledges in the South and maybe Rob Lucas, if he's in, at King George, Ilford, may have one too. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Thursday, February 16, 2006 4:04 PM To: Kemlo Rogerson; Nancy Lemke; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] whole brain in paraffin/microtome for sale? Talking of tetranders, has anybody in the UK got a decent sledge microtome to sell, preferably for a song. (I can do guitar accompaniment if necessary)? Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:kemlo.rogerson@waht.swest.nhs.uk] Sent: 16 February 2006 14:18 To: 'Nancy Lemke'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] whole brain in paraffin WOW a Tetrander!!! Thought those things had been consigned to the torture chamber..... Used that thing 25 years ago with mega brain slices, hardest bit was keeping the block cool but as long as the processing was Ok (used to take weeks to process the brain) it was not too bad; needed skill in manipulating the sections and then floating onto dirty great big coated slides. If you have any probs then contact me. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Nancy Lemke [mailto:nsnwl@neuro.hfh.edu] Sent: Thursday, February 16, 2006 1:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] whole brain in paraffin Good Morning Histonet, I would like some information about the preparation and sectioning of 1-2cm slabs of human brain. The cutting will be done on an old Reichert Tetrander (sliding) microtome with a giant steel blade. If anyone who has experience with this sort of sectioning would email me I would love to ask some questions. Thank you in advance, Nancy Lemke Research Coordinator Hermelin Brain Tumor Center Henry Ford Hospital Detroit ============================================================================ == Go to http://henryford.com We're Henry Ford. We Can. HFHS CONFIDENTIALITY NOTICE: This email contains information from the sender that may be CONFIDENTIAL, LEGALLY PRIVILEGED, PROPRIETARY or otherwise protected from disclosure. This email is intended for use only by the person or entity to whom it is addressed. If you are not the intended recipient, any use, disclosure, copying, distribution, printing, or any action taken in reliance on the contents of this email, is strictly prohibited. If you received this email in error, please contact the sending party by replying in an email to the sender, delete the email from your computer system and shred any paper copies of the email you printed. Note to Patients: There are a number of risks you should consider before using e-mail to communicate with us. These risks are described in our Privacy Policy at http://henryford.com. Review that policy carefully before continuing to communicate with us by e-mail. For greater Internet security, our policy describes the Henry Ford MyHealth electronic communication process - you may register at http://henryford.com. If you do not believe that our policy gives you the privacy and security protection you need, do not send e-mail or Internet communications to us. ============================================================================ == _______________________________________________ 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 From Tracey.Lenek <@t> CLS.ab.ca Thu Feb 16 10:39:29 2006 From: Tracey.Lenek <@t> CLS.ab.ca (Tracey.Lenek@CLS.ab.ca) Date: Thu Feb 16 10:39:41 2006 Subject: [Histonet] Renal biopsies washing off slides Message-ID: <4BC300747AF87A48BCDF8E48BC2885CE02C1D4BF@mail1.calgary.com> We are having a transient issue with renal biopsies washing off coated slides. They are all processed in B5 from VWR and diluted with formalin from Fisher and a small amount of safranin to color the tissue. Tissue from yesterday appeared shrunken and brittle - but biopsies from previous days were fine using above reagents from the same lot number. Any experience or thoughts regarding this issue would be appreciated. Thanks, Tracey CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify the sender immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. From NMargaryan <@t> childrensmemorial.org Thu Feb 16 11:51:32 2006 From: NMargaryan <@t> childrensmemorial.org (Margaryan, Naira) Date: Thu Feb 16 11:51:04 2006 Subject: [Histonet] RE: Histonet Digest Message-ID: <63B8B599DE283148B92E83C78B32C15D018A7CCD@cmhexbe2.childrensmemorial.org> Good Morning Histonet, I have a question regarding chick embryo and frizzing them. Could you, please give me any suggestion: How to freeze chick embryo properly? The chick embryo is a bit different tissue. Any help would be appreciated.... Naira Naira V. Margaryan, D.V.M., Ph.D. Research Associate III Children's Memorial Research Center 2300 Children's Plaza, Box 222 Chicago, IL 60614-3394 Tel: 773-755-6570/ext-8 Fax: 773-755-6594 nmargaryan@childrensmemorial.org For Express Mail: CMRC, Room C.473 2430 N. Halsted Street Chicago, IL 60614-4314 From mohana_g2002 <@t> yahoo.com Thu Feb 16 12:19:18 2006 From: mohana_g2002 <@t> yahoo.com (MOHANA) Date: Thu Feb 16 12:19:26 2006 Subject: [Histonet] cryostat sections Message-ID: <20060216181918.45982.qmail@web33505.mail.mud.yahoo.com> hello all!! i was just wondering if someone can tell me how do we fix the cryostat sections... as i have seen that since the sections r already stored at a low temp , fixing them in carnoys fixative kept at room temp..resulte in shrinking of cells!!! mohana Mohana --------------------------------- Yahoo! Autos. Looking for a sweet ride? Get pricing, reviews, & more on new and used cars. From straussj <@t> upstate.edu Thu Feb 16 12:33:14 2006 From: straussj <@t> upstate.edu (Judy Strauss) Date: Thu Feb 16 12:33:51 2006 Subject: [Histonet] laser microdissection /rat bones Message-ID: Hello Histonet, One of our residents wants to study gene expression of primary osteoblasts from rat long bones using laser dissection and microarray analysis. Does anyone in Histoland have experience with this type of animal model? Any help will be gratefully accepted. These techniques are so far outside my area of expertise I do not even know what questions to ask. Thanks, Judith Strauss SUNY Upstate Medical University Department of Orthopedic Surgery IHP room 3118 505 Irving Avenue Syracuse, NY 13120 phone: (315) 464-9960 fax: (315) 464-6638 From TJJ <@t> Stowers-Institute.org Thu Feb 16 12:43:09 2006 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Thu Feb 16 12:43:29 2006 Subject: [Histonet] Re: Chick embryo freezing Message-ID: Naira, We will formalin fix and cryopreserve chick embryos before freezing in OCT with good results. What sorts of issues are you having? What age sample do you have? Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From clcses <@t> gmail.com Thu Feb 16 12:54:07 2006 From: clcses <@t> gmail.com (Carmen Contreras-Sesvold) Date: Thu Feb 16 12:54:14 2006 Subject: [Histonet] need to buy a cryostat, advice please Message-ID: <46a3be380602161054w14b29b3du93939de53c256cbc@mail.gmail.com> I am looking to purchase a new cryostat. I want one with a small foot print (takes up little floor space) that can adjust and maintain the temperature of the head as well as the chamber. I will be using it to cut frozen rodent tissue (lung and brain) thicknesses from 5 to 100 microns. Have any of you purchase something like this recently, can you offer any advice? Has anyone used an newly purchased cryostat and can offer a reccommendation??? Thanks so much. Carmen From dpahisto <@t> yahoo.com Thu Feb 16 13:17:55 2006 From: dpahisto <@t> yahoo.com (Cindy DuBois) Date: Thu Feb 16 13:18:06 2006 Subject: [Histonet] Pneumocystis controls Message-ID: <20060216191756.49285.qmail@web33409.mail.mud.yahoo.com> I am in need of a good pneumocystis control. We have bought our last 3 batches from vendors and our pathologists are not happy with them. I would be happy to trade another control (??). Cindy DuBois Integrated Pathology Stockton, CA --------------------------------- Brings words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. From cpomajzl <@t> cpllabs.com Thu Feb 16 13:23:04 2006 From: cpomajzl <@t> cpllabs.com (Chris Pomajzl) Date: Thu Feb 16 13:21:08 2006 Subject: [Histonet] laser microdissection /rat bones References: Message-ID: <00c701c6332e$694b1bc0$26fca8c0@CSP> Contact Dr. Jim Richardson or John Shelton with the Molecular Pathology Core at UT Southwestern Medical Center at Dallas. They should be able to give you some information. ----- Original Message ----- From: "Judy Strauss" To: Sent: Thursday, February 16, 2006 12:33 PM Subject: [Histonet] laser microdissection /rat bones > Hello Histonet, > > One of our residents wants to study gene expression of primary osteoblasts > from rat long bones using laser dissection and microarray analysis. Does > anyone in Histoland have experience with this type of animal model? > > Any help will be gratefully accepted. These techniques are so far outside > my area of expertise I do not even know what questions to ask. > > Thanks, > > > > Judith Strauss > SUNY Upstate Medical University > Department of Orthopedic Surgery > IHP room 3118 > 505 Irving Avenue > Syracuse, NY 13120 > > phone: (315) 464-9960 > fax: (315) 464-6638 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jlinda <@t> ces.clemson.edu Thu Feb 16 15:13:26 2006 From: jlinda <@t> ces.clemson.edu (Linda Jenkins) Date: Thu Feb 16 15:09:47 2006 Subject: [Histonet] Re: PLGA frozen section problem Message-ID: <5.2.1.1.2.20060216160552.01edbbc0@mailhost.ces.clemson.edu> Ashwin, All polymeric scaffolds are NOT equal! I have found that using the Instrumedics tape transfer technique really helps with the frozen PLLA scaffolds. In fact I have a PowerPoint presentation that shows the step-by-step technique.Cutting a frozen section on a PLLA scaffold without the tape technique is like trying to section a loosely held piece of dust...you sneeze...it's gone! Please email me if you would like the slide show. Good Luck, Linda Linda Jenkins, HT Clemson University Dept. of Bioengineering Clemson, SC 29634-0905 864.656.5553 http://www.ces.clemson.edu/bio/research/histo.htm From gentras <@t> vetmed.auburn.edu Thu Feb 16 15:31:16 2006 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Thu Feb 16 15:31:30 2006 Subject: Fwd: [Histonet] OCT embedding of NBF-fixed specimen Message-ID: <6.0.1.1.0.20060216152351.01999e38@mailhost.vetmed.auburn.edu> Hello, we usually cryoprotect in 10%, 20%, & 30% Sucrose respectively in either the fixative which the sample was fixed or in 0.1M Phosphate Buffer. It really depends on the staining technique you plan to use. Allow tissue sample to sink in each percentage of sucrose before transferring, and after sample sinks in 30% it's usually safe to freeze. This technique has worked well for us yielding little or no ice crystal artifact. Best wishes. Atoska >DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; s=s1024; d=yahoo.com; > >h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; > >b=hyDJcagLptAfKqyzRODPc93k0ZWR9p1W0SLjM7b7xQnv3pQgQleX7KL4DRHQ8YCaie0ANpCPFqNSM2DbTCV/GpIvz81FfeXM02B8gx+FvIIgx925bp5wWDRa1iZQ3LgX4vFSrOtLVhWMUwm6O+VPShqmqHEgy7oMJG+/EXCLjEs= > ; >Date: Thu, 16 Feb 2006 06:27:53 -0800 (PST) >From: Kim Merriam >To: Histonet >X-Scan-Signature: 218f69a45ba25cab82cec6de17a14311 >X-Content-Filtered-By: Mailman/MimeDel 2.1.5 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.5 >List-Id: For the exchange of information pertaining to histotechnology and > related fields >List-Unsubscribe: >, > > >List-Archive: >List-Post: >List-Help: >List-Subscribe: , > >Sender: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: ab3f4c55490e4d672805895add812bc5 >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >Subject: [Histonet] OCT embedding of NBF-fixed specimen >X-Spam-Checker-Version: SpamAssassin 2.64 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: *** >X-Spam-Status: No, hits=3.2 required=5.5 tests=FROM_ENDS_IN_NUMS,RCVD_IN_DSBL, > RCVD_IN_NJABL_PROXY autolearn=no version=2.64 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >X-spam: ESP<-54>=RBL:<-140> SHA:<37> UHA:<0> SLS:<0> BAYES:<49> SenderID:<0> > Spam Dictionary (TRU10):<0> Obscenities Dictionary > (TRU10):<0> Scam Dictionary (TRU10):<0> Adult Dictionary > (TRU10):<0> Embed HTML Dictionary (TRU10):<0> Float > Dictionary (TRU10):<0> HTML Dictionary (TRU10):<0> URL > Real-Time Signatures:<0> Spam Dictionary 2 (TRU10):<0> > >Hello all, > > I need to do some OCT embedding of tissues that are already fixed in > formalin. What is the best way to do this? Should I rinse the tissues > in water to get out the formalin? How can I prevent crystal artifact on > these tissues? > > Any advice would be helpful, > > Thanks, > Kim > > >Kim Merriam >Novartis >Cambridge, MA > >--------------------------------- >Brings words and photos together (easily) with > PhotoMail - it's free and works with Yahoo! Mail. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jlinda <@t> ces.clemson.edu Thu Feb 16 16:23:37 2006 From: jlinda <@t> ces.clemson.edu (Linda Jenkins) Date: Thu Feb 16 16:19:58 2006 Subject: [Histonet] Re: High density slide storage Message-ID: <5.2.1.1.2.20060216164157.01ff9778@mailhost.ces.clemson.edu> Leslie, Southern Business Systems, Inc. is one of the companies that does automated materials management. It is really a neat system. Slides are barcoded before filing and can be retrieved automatically. Their website is http://www.sbssolutions.com/03_products/auto.shtml It shows a picture of pathology slides and lab samples. I have one of their old brochures and it says they put five years of histology slides (very large hospital) into 16 square feet of floor space. Hope this was what you were thinking about, Linda Linda Jenkins, HT Clemson University Dept. of Bioengineering Clemson, SC 29634-0905 864.656.5553 http://www.ces.clemson.edu/bio/research/histo.htm From Adesupod <@t> aol.com Thu Feb 16 16:43:26 2006 From: Adesupod <@t> aol.com (Adesupod@aol.com) Date: Thu Feb 16 16:43:46 2006 Subject: [Histonet] Tissue Specimens Message-ID: <1dc.4e8151bb.31265a0e@aol.com> Hi, Pls I will appreciate it, If I could be able to get the following tissue specimens listed below for my colleague who is preparing for ASCP. The tissue specimens are; 1. UTERUS 2. INTESTINE 3. CEREBELLUM Thanking you all for your usual cooperation. Adesupo Adesuyi. From lpwenk <@t> sbcglobal.net Thu Feb 16 19:24:37 2006 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Thu Feb 16 19:25:11 2006 Subject: [Histonet] Tissue Specimens In-Reply-To: <1dc.4e8151bb.31265a0e@aol.com> Message-ID: <000a01c63360$eb9bc890$4d29d445@HPPav2> We need a little more information: Size of tissue What other requirements (endometrium, myometerium, layers, etc.) Is that small or large intestine? To whom do we send it? Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Adesupod@aol.com Sent: Thursday, February 16, 2006 5:43 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tissue Specimens Hi, Pls I will appreciate it, If I could be able to get the following tissue specimens listed below for my colleague who is preparing for ASCP. The tissue specimens are; 1. UTERUS 2. INTESTINE 3. CEREBELLUM Thanking you all for your usual cooperation. Adesupo Adesuyi. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From great_scientist <@t> yahoo.com Thu Feb 16 23:41:52 2006 From: great_scientist <@t> yahoo.com (cheng-chi chao) Date: Thu Feb 16 23:42:00 2006 Subject: [Histonet] Trap staining Message-ID: <20060217054152.83532.qmail@web33208.mail.mud.yahoo.com> Can anyone help me out for Trap staining? There are two kits available from Sigma (386 and 387). Which one work for formalin-fixed, EDTA decal, paraffin-embedded mouse limbs? Thanks very mcuh for your help in advance. CC __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From angela.mcnabola.b <@t> bayer.com Fri Feb 17 06:47:06 2006 From: angela.mcnabola.b <@t> bayer.com (Angela McNabola) Date: Fri Feb 17 06:46:37 2006 Subject: [Histonet] Trap staining In-Reply-To: <20060217054152.83532.qmail@web33208.mail.mud.yahoo.com> Message-ID: See our protocl for TRAP below. I have to be honest, I had to get this one out of the archives since we don't really do it anymore. It works quite nicely though. I can send you a picture, but contact me directly if interested. Angela Sigma Diagnostics Cat. #387-A Leukocyte Acid Phosphatase Kit Bones are cold fixed in 10% NBF, and decalcified in a 5% EDTA solution (cold) over several days-weeks depending upon sample size. Kit Includes: 10ml Napthol AS-BI Phosphoric Acid Solution 10ml Fast Garnet GBC Base Solution 10ml Tartrate Solution 50ml Acetate Solution 10ml Sodium Nitrate Solution 50ml Citrate Solution 50ml Hematoxylin Solution, Gill No. 3 1. Deparaffinize slides to water 2. For approximately 5 Coplin jars (48 slides) n 270 ml distilled water (warm) n 3ml Nitrate Solution n 3ml Fast Garnet Solution ( Note: add Nitrate Solution and Fast Garnet together 1 minute prior to adding all solutions together) n 12mls acetate 6ml Tartrate Solution n 3ml Napthol Solution 3. Mix all reagents together and pour in Coplin jars 4. Place Coplin jars in waterbath incubator for approximately 45 minutes to 1 hour 5. Rinse in warm water (2 rinses) 6. Place slides in Weigert?s Hematoxylin (mix parts A and B equally) for 2 minutes 7. Rinse in warm water (2 rinses) 8. Place slides in filtered .025% aqueous Light Green Solution for 10 seconds 9. Quick rinse in 95% ETOH then water 10. Aqueous mount Angela McNabola MS, HT(ASCP)SLS, QIHC Scientist Bayer Healthcare 400 Morgan Lane West Haven, CT 06516 203-812-5001 fax# 203-812-5820 angela.mcnabola.b@bayer.com cheng-chi chao To: histonet@lists.utsouthwestern.edu Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] Trap staining western.edu 02/17/2006 12:41 AM Can anyone help me out for Trap staining? There are two kits available from Sigma (386 and 387). Which one work for formalin-fixed, EDTA decal, paraffin-embedded mouse limbs? Thanks very mcuh for your help in advance. CC __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________________________________________________________ The information contained in this e-mail is for the exclusive use of the intended recipient(s) and may be confidential, proprietary, and/or legally privileged. Inadvertent disclosure of this message does not constitute a waiver of any privilege. If you receive this message in error, please do not directly or indirectly use, print, copy, forward, or disclose any part of this message. Please also delete this e-mail and all copies and notify the sender. Thank you. For alternate languages please go to http://bayerdisclaimer.bayerweb.com _______________________________________________________________________________________________ From MSafron <@t> wilresearch.com Fri Feb 17 07:13:49 2006 From: MSafron <@t> wilresearch.com (Mike Safron) Date: Fri Feb 17 07:17:00 2006 Subject: [Histonet] Testes - Fixation, Staining, Staging Message-ID: Hello, I read the following topic on the histonet and had some questions. Do you find that prolonged fixation in Bouins > 24 hrs causes a decrease in acrosomal staining in the testes? Thanks, Michael Safron A.S.,HT(ASCP) Manager, Histology WIL Research Laboratories LLC 419-289-8700 ext: 3040 Ashland, Ohio 44805 Hello, Here instructions for HE and PAS after Hotchkiss my Pathologists likes;) We fix testis in Boiun for 24 hours, then transfer them to 70% ethanol in which they can be kept as long as you wish. We do not rinse them several times in 70% ethanol. If you wish I can ask him more about satging when he returns from Christmas vacation. Hope this helpes you:) I can send you also pictures of my stainings if you like to see the result. Bettina Hutz Research Assistant Department of Discovery Biology Orion Corporation, ORION PHARMA Orionintie 1, P.O.Box 65, FIN-02101 Espoo, Finland Tel. +358 10 429 2607, Fax +358 10 429 2924 From cpomajzl <@t> cpllabs.com Fri Feb 17 07:51:47 2006 From: cpomajzl <@t> cpllabs.com (Chris Pomajzl) Date: Fri Feb 17 07:49:56 2006 Subject: [Histonet] Testes - Fixation, Staining, Staging References: Message-ID: <003801c633c9$4bc5b8d0$26fca8c0@CSP> Bouin's is acidic, so I would think that prolonged fixation could adversely affect DNA/RNA, acrosomal staining. We would fix in Bouin's for 4-6 hours and then transfer to 70% alcohol. ----- Original Message ----- From: "Mike Safron" To: Sent: Friday, February 17, 2006 7:13 AM Subject: [Histonet] Testes - Fixation, Staining, Staging Hello, I read the following topic on the histonet and had some questions. Do you find that prolonged fixation in Bouins > 24 hrs causes a decrease in acrosomal staining in the testes? Thanks, Michael Safron A.S.,HT(ASCP) Manager, Histology WIL Research Laboratories LLC 419-289-8700 ext: 3040 Ashland, Ohio 44805 Hello, Here instructions for HE and PAS after Hotchkiss my Pathologists likes;) We fix testis in Boiun for 24 hours, then transfer them to 70% ethanol in which they can be kept as long as you wish. We do not rinse them several times in 70% ethanol. If you wish I can ask him more about satging when he returns from Christmas vacation. Hope this helpes you:) I can send you also pictures of my stainings if you like to see the result. Bettina Hutz Research Assistant Department of Discovery Biology Orion Corporation, ORION PHARMA Orionintie 1, P.O.Box 65, FIN-02101 Espoo, Finland Tel. +358 10 429 2607, Fax +358 10 429 2924 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tonyc <@t> uhnresearch.ca Fri Feb 17 08:36:31 2006 From: tonyc <@t> uhnresearch.ca (Tony Collins) Date: Fri Feb 17 08:36:44 2006 Subject: [Histonet] Microscopy Today mounting media article In-Reply-To: <20060216180111.9E745612@mail0.uhnres.utoronto.ca> References: <20060216180111.9E745612@mail0.uhnres.utoronto.ca> Message-ID: <43F5DF6F.7070503@uhnresearch.ca> I'd first like to thank people for the kind feedback I've had regarding my article in Microscopy Today. I'm pleased people are finding it useful. I certainly found compiling it very helpful. The article started off as kind of bullet-point document detailing the experiences of the confocal and histonet users. It was originally a PDF on our website before Microscopy Today approached me for a more polished, expanded version. It was always meant to be a 'work in progress' continually updated as users gain more experience with the increasing number of mounting media. The online version will be updated periodically. There is currently an interesting thread on the confocal-list discussing how to mount live Paramecium! You can download the latest PDF of the article from our website: http://www.uhnresearch.ca/facilities/wcif/fdownload2.html The big advantage of the PDF version is that it's 'updateable'. There are also clickable-links in the PDF to the original archive-comments for those who want to dig further. Kind regards, Tony -- Tony Collins, Ph.D. Facility Manager Wright Cell Imaging Facility Toronto Western Research Institute 13-407 McLaughlin Pavilion 399 Bathurst Street Toronto, ON. M5T 2S8 tel. (416) 603 5367 fax: (416) 603 5745 http://www.uhnresearch.ca/wcif From meint002 <@t> umn.edu Fri Feb 17 11:20:42 2006 From: meint002 <@t> umn.edu (meint002) Date: Fri Feb 17 11:20:49 2006 Subject: [Histonet] Pre-treatment module for immunostainers Message-ID: <200602171720.k1HHKhgF011928@vanguard.software.umn.edu> Dear Histos, I am in the process of determining which immunostainer to purchase for our lab. DAKO and Lab Vision are the main vendors at the moment. Lab Vision has a new device that they claim will automate the dewaxing and epitope recovery on your slide before you apply the antibodies. Does anyone have experience with this Pre-treatment module or a device of a similar nature? If so, do they work very well, or are they worth the expense? I think you can use your own reagents for the various buffers - so that could keep the reagent cost down some. Otherwise, they would be happy to sell you their own products. Any advice on this matter would be greatly appreciated. Thank you in advance for your feedback. Joyce Meints Histologist University of Minnesota Paul and Sheila Wellstone Muscular Dystrophy Center MMC 206 420 Washington St. SE Minneapolis, MN 55455-0392 e-mail: meint002@umn.edu lab phone: 612-626-4703 From Jeannette.Mitchell <@t> vtmednet.org Fri Feb 17 11:32:29 2006 From: Jeannette.Mitchell <@t> vtmednet.org (Mitchell, Jeannette M.) Date: Fri Feb 17 11:32:39 2006 Subject: [Histonet] (no subject) Message-ID: Hello Folks, We're looking for a digital dictation system for our grossing area to replace our dictaphones and tapes. We use CoPath for our pathology information system. Any suggestions ? thanks ! Jeannette Mitchell FAHC Burlington,VT Confidentiality Notice: This message, and any attachments, may contain information that is confidential, privileged, and/or protected from disclosure under state and federal laws that deal with the privacy and security of medical information. If you received this message in error or through inappropriate means, please reply to this message to notify the Sender that the message was received by you in error, and then permanently delete this message from all storage media, without forwarding or retaining a copy. From rjbuesa <@t> yahoo.com Fri Feb 17 11:49:57 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Feb 17 11:50:04 2006 Subject: [Histonet] Pre-treatment module for immunostainers In-Reply-To: <200602171720.k1HHKhgF011928@vanguard.software.umn.edu> Message-ID: <20060217174957.75639.qmail@web61211.mail.yahoo.com> Dear Joyce: Unless you are short in trained personnel and perform more than 100 IHC slides daily, I personally do not see the "advantage" of limiting yourself to a company that will sell you proprietary (and expensive) reagents that will lead you to a "closed system". "Ventana" has a new IHC automatic instrument that also deparafinizes and performs HIER and I have heard both good and bad things about it. I always used Dako and laboratories that I have consulted for also use Dako (up to 3 autostainers) and the "beautity" of the Dako instrument is that it is an "open" technology that you can use even with home made reagents (HIER buffers, Ab diluents) as well as any Ab from any manufacturer. To dewax your slides will take less than 30 minutes (and it can be done with your regular H&E autostainer) and HIER requires, at the most, 1 hour, and you don't have to stand by during that time. I would not commit myself to a single manufacturer product. I hope that this will help you! Ren? J. meint002 wrote: Dear Histos, I am in the process of determining which immunostainer to purchase for our lab. DAKO and Lab Vision are the main vendors at the moment. Lab Vision has a new device that they claim will automate the dewaxing and epitope recovery on your slide before you apply the antibodies. Does anyone have experience with this Pre-treatment module or a device of a similar nature? If so, do they work very well, or are they worth the expense? I think you can use your own reagents for the various buffers - so that could keep the reagent cost down some. Otherwise, they would be happy to sell you their own products. Any advice on this matter would be greatly appreciated. Thank you in advance for your feedback. Joyce Meints Histologist University of Minnesota Paul and Sheila Wellstone Muscular Dystrophy Center MMC 206 420 Washington St. SE Minneapolis, MN 55455-0392 e-mail: meint002@umn.edu lab phone: 612-626-4703 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! From p.rastogi <@t> biogenex.com Fri Feb 17 12:16:25 2006 From: p.rastogi <@t> biogenex.com (Promila Rastogi) Date: Fri Feb 17 12:16:34 2006 Subject: [Histonet] RE: Pre-treatment module for immunostainers Message-ID: <37DC9F93CF7F864182D0463EF93D571B0C182E@ISLETON2.california.biogenex.com> FYI, BioGenex has a pretreatment system called the EZ Retriever. It is a temperature and time controlled microwave based system and can treat up to 96 slides in one run (each run is no more than 10 minutes). You may use your own Antigen Retrieval solutions (which are aqueous), or BioGenex's. Decide what temperature you want to treat your slides at. Promila Rastogi, Product Manager 4600 Norris Canyon Road San Ramon, CA 94583 Tel: 925.866-2577 Fax: 925.866-2594 ================================================================================ Message: 19 Date: Fri, 17 Feb 2006 09:49:57 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Pre-treatment module for immunostainers To: meint002 , histonet@lists.utsouthwestern.edu Message-ID: <20060217174957.75639.qmail@web61211.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Dear Joyce: Unless you are short in trained personnel and perform more than 100 IHC slides daily, I personally do not see the "advantage" of limiting yourself to a company that will sell you proprietary (and expensive) reagents that will lead you to a "closed system". "Ventana" has a new IHC automatic instrument that also deparafinizes and performs HIER and I have heard both good and bad things about it. I always used Dako and laboratories that I have consulted for also use Dako (up to 3 autostainers) and the "beautity" of the Dako instrument is that it is an "open" technology that you can use even with home made reagents (HIER buffers, Ab diluents) as well as any Ab from any manufacturer. To dewax your slides will take less than 30 minutes (and it can be done with your regular H&E autostainer) and HIER requires, at the most, 1 hour, and you don't have to stand by during that time. I would not commit myself to a single manufacturer product. I hope that this will help you! Ren? J. meint002 wrote: Dear Histos, I am in the process of determining which immunostainer to purchase for our lab. DAKO and Lab Vision are the main vendors at the moment. Lab Vision has a new device that they claim will automate the dewaxing and epitope recovery on your slide before you apply the antibodies. Does anyone have experience with this Pre-treatment module or a device of a similar nature? If so, do they work very well, or are they worth the expense? I think you can use your own reagents for the various buffers - so that could keep the reagent cost down some. Otherwise, they would be happy to sell you their own products. Any advice on this matter would be greatly appreciated. Thank you in advance for your feedback. Joyce Meints Histologist University of Minnesota Paul and Sheila Wellstone Muscular Dystrophy Center MMC 206 420 Washington St. SE Minneapolis, MN 55455-0392 e-mail: meint002@umn.edu lab phone: 612-626-4703 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 27, Issue 27 **************************************** From jcresor <@t> lcpath.com Fri Feb 17 12:25:54 2006 From: jcresor <@t> lcpath.com (Jennifer N. Cresor) Date: Fri Feb 17 12:26:11 2006 Subject: [Histonet] catscratch immuno Message-ID: <200602171826.k1HIQ1p13045@plus34.host4u.net> Hi, Our lab has purchased a catscratch antibody. We purchased a control with Bartonella henselae organisms. The control slide stains positive in several areas therefore not being able to see the organisms. We have tried patient tissue also with no luck. Does anyone out there have some ideas to help get a control that works for us? Thanks! Jennifer jcresor@lcpath.com From Rcartun <@t> harthosp.org Fri Feb 17 12:43:38 2006 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Feb 17 12:44:30 2006 Subject: [Histonet] catscratch immuno Message-ID: Bartonella henselae are difficult to detect. Unless you have significant experience detecting "bugs" with IHC, and see frequent cases, you are going to have problems. The presence of bugs will depend on the duration of the disease, prior treatment with antibiotics, and sampling. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Jennifer N. Cresor" 02/17/06 01:25PM >>> Hi, Our lab has purchased a catscratch antibody. We purchased a control with Bartonella henselae organisms. The control slide stains positive in several areas therefore not being able to see the organisms. We have tried patient tissue also with no luck. Does anyone out there have some ideas to help get a control that works for us? Thanks! Jennifer jcresor@lcpath.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mauger <@t> email.chop.edu Fri Feb 17 13:18:26 2006 From: mauger <@t> email.chop.edu (Joanne Mauger) Date: Fri Feb 17 13:19:23 2006 Subject: [Histonet] catscratch immuno Message-ID: Jennifer, I tried in vain for many months to get a good tissue control. I used the rat lung commercially available from Newcomer Supply to optimize. If you think you see positive staining, look at 100X, under oil if need be to confirm the tiny, rod shaped organism. Sometimes they look round. A red chromogen may help you see it better. The docs weren't convinced until I stained a case that had nice scattered positivity. Now I guard that case with my life for control tissue, but the rat lung stains positively every time. Look for old cases that were suspect for cat scratch,where the serum tested positive, and stain the tissue. Hopefully you'll get lucky. I use a 1:50 dilution with no retrieval necessary.It has worked well with ABC, Dakos envision+, LSAB. Jo Mauger >>> "Jennifer N. Cresor" 02/17/06 1:25 PM >>> Hi, Our lab has purchased a catscratch antibody. We purchased a control with Bartonella henselae organisms. The control slide stains positive in several areas therefore not being able to see the organisms. We have tried patient tissue also with no luck. Does anyone out there have some ideas to help get a control that works for us? Thanks! Jennifer jcresor@lcpath.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From NMargaryan <@t> childrensmemorial.org Fri Feb 17 14:18:08 2006 From: NMargaryan <@t> childrensmemorial.org (Margaryan, Naira) Date: Fri Feb 17 14:22:16 2006 Subject: [Histonet] Chick embryo freezing Message-ID: <63B8B599DE283148B92E83C78B32C15D018A7DBD@cmhexbe2.childrensmemorial.org> Thanks Teri, Problem is: I have to fix/cut or cut/fix the chick embryos with GFP labeled cells into; and then use these fluorescent labeled cells sections for the laser capture microdissection and microarray analysis. This technique is a new for us. I did send my question to Arcturus Company and do not have any answer yet. Any help will be gratefully accepted, Naira Naira V. Margaryan, D.V.M., Ph.D. Research Associate III Children's Memorial Research Center 2300 Children's Plaza, Box 222 Chicago, IL 60614-3394 Tel: 773-755-6570/ext-8 Fax: 773-755-6594 nmargaryan@childrensmemorial.org ------------------------------ Naira, We will formalin fix and cryopreserve chick embryos before freezing in OCT with good results. What sorts of issues are you having? What age sample do you have? Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From plaurie <@t> benaroyaresearch.org Fri Feb 17 15:14:45 2006 From: plaurie <@t> benaroyaresearch.org (Patrick Laurie) Date: Fri Feb 17 15:14:54 2006 Subject: [Histonet] 20 micron sections Cresyl Violet Acetate stained Message-ID: <632ae8e2e68de84580523eede7cb1009@penguin.vmresearch.org> Hi Histonet, My group is interested in doing an anterior horn alpha motor neuron count of ALS pts vs control. There is considerable literature about these techniques, which has been most helpful, but the devil is in the details. Most papers use 20um sections of the ventral horn stained with cresyl violet acetate, or iron-chromoxane cyanine R method for myelin (counterstained with .5% neutral red) (Dr. Kiernan's 1991 paper), or one of many other methods for staining. I have 3 questions. First of all, a technical question, how are 20um sections kept on the slides during staining? I have done some experimenting, and found it very difficult to keep them on. Secondly, how do the thick sections stain? The cresyl violet using will not differentiate very well (using the regressive version), and the H&E is too dark, I have yet to use any other method. And lastly, why 20um? Conventional neural histology seems to be done thicker, but usually not that thick. What would be the benefit for having sections that thick? Thanks Histonet, Patrick Laurie, HT (ASCP) Neurogenomics Laboratory Benaroya Research Institute 1201 9th Ave Seattle, Wa 98101 From jnocito <@t> satx.rr.com Fri Feb 17 16:46:18 2006 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Feb 17 16:46:28 2006 Subject: [Histonet] (no subject) References: Message-ID: <001801c63413$f7f6d6d0$e8bd0b43@yourxhtr8hvc4p> Jeanette, we use Fusion. It is hooked up to our computer. We have foot pedals like Dictaphone, but when we dictate, it goes on our computer. The transcriptionists pull out the dictations and transcribe into our homemade LIS, which is not Copath, but should work with any system. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Mitchell, Jeannette M." To: Sent: Friday, February 17, 2006 11:32 AM Subject: [Histonet] (no subject) Hello Folks, We're looking for a digital dictation system for our grossing area to replace our dictaphones and tapes. We use CoPath for our pathology information system. Any suggestions ? thanks ! Jeannette Mitchell FAHC Burlington,VT Confidentiality Notice: This message, and any attachments, may contain information that is confidential, privileged, and/or protected from disclosure under state and federal laws that deal with the privacy and security of medical information. If you received this message in error or through inappropriate means, please reply to this message to notify the Sender that the message was received by you in error, and then permanently delete this message from all storage media, without forwarding or retaining a copy. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- No virus found in this incoming message. Checked by AVG Free Edition. Version: 7.1.375 / Virus Database: 267.15.10/263 - Release Date: 2/16/2006 From tissuearray <@t> hotmail.com Fri Feb 17 16:47:20 2006 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Feb 17 16:47:29 2006 Subject: [Histonet] Microarray Message-ID: Hey Lin, Which arrayer are you using? If you are using the Chemicon arrayer, their recipient needle is slightly bigger then the donor needle cores. You will need to set the depth of each recipient hole to match the donor core. If you are using the Beecher Instrument, then the donor core will fit snuggly in the Recipient hole unless you made the hole larger then the original hole on accident. Which often happens when people are learning to punch. If you are loosing cores from the array block while sectioning the array block on a microtome, then the problem is you haven't set the punches well. Put the finished array block in an oven on a glass slide (punch surface down on slide) at around 37 to 40 degrees Celsius for 15 minutes then press the glass slide and block together. This smashes the paraffin around the cores so the cores stick to the paraffin in the array block. If you don't do this step before you sections the array block the punches will fall out. This step is for both array instruments. I hope this helps, Cheers, Thom Jensen For more information of array instruction visit: www.arrayworkshop.com > >Hi, >Are there any hints to share on getting all the cores to stay evenly in the >recipient block and to get sections without losing cores? >Lin >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Fri Feb 17 18:11:16 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Feb 17 18:12:00 2006 Subject: [Histonet] (no subject) References: <001801c63413$f7f6d6d0$e8bd0b43@yourxhtr8hvc4p> Message-ID: <83AACDB0810528418AA106F9AE9B7F7E018252BE@sjhaexc02.sjha.org> WE use Lanier which is very satisfactory for us. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Joe Nocito Sent: Fri 2/17/2006 5:46 PM To: Mitchell, Jeannette M.; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] (no subject) Jeanette, we use Fusion. It is hooked up to our computer. We have foot pedals like Dictaphone, but when we dictate, it goes on our computer. The transcriptionists pull out the dictations and transcribe into our homemade LIS, which is not Copath, but should work with any system. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Mitchell, Jeannette M." To: Sent: Friday, February 17, 2006 11:32 AM Subject: [Histonet] (no subject) Hello Folks, We're looking for a digital dictation system for our grossing area to replace our dictaphones and tapes. We use CoPath for our pathology information system. Any suggestions ? thanks ! Jeannette Mitchell FAHC Burlington,VT Confidentiality Notice: This message, and any attachments, may contain information that is confidential, privileged, and/or protected from disclosure under state and federal laws that deal with the privacy and security of medical information. If you received this message in error or through inappropriate means, please reply to this message to notify the Sender that the message was received by you in error, and then permanently delete this message from all storage media, without forwarding or retaining a copy. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- No virus found in this incoming message. Checked by AVG Free Edition. Version: 7.1.375 / Virus Database: 267.15.10/263 - Release Date: 2/16/2006 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From Adesupod <@t> aol.com Fri Feb 17 20:40:37 2006 From: Adesupod <@t> aol.com (Adesupod@aol.com) Date: Fri Feb 17 20:40:57 2006 Subject: [Histonet] Tissue Specimens Needed for ASCP Message-ID: <237.71944cb.3127e325@aol.com> ? Hi, ??? Pls I will appreciate it, If I can be able to get the following tissue specimens listed below for my colleague ASCP test. 1. SMALL INTESTINE????? - 2.0cm in length ?????????????????????????????????????? - to include all layers ?????????????????????????????????????? - Epithelium to cover one entire surface and not autolized. 2.CEREBELLUM???????????? -? 1.0 X 1.0 cm ????????????????????????????????????? -? to include grey and white matter. 3. SKELETAL MUSCLE? - 1.0 x 1.0cm ????????????????????????????????????? - Longitudinal section. 4. UTERUS?????????????? -? 1.5 x 1.5cm ????????????????????????????????? -? To include endometrium and myometrium. ????????????????????????????????? -?? Endometrium to cover one entire surface.. ????? MY ADDRESS: ???????????????????????????????? ADESUPO ADESUYI ????????????????????????????????? HISTOLOGY DEPT. ???????????????????????????????? LOVELACE? MEDICAL CENTER, ???????????????????????????????? 5400 GIBSON BLVD. SE ??????????????????????????????? ALBUQUERQUE, NM 87108. From kemlo <@t> f2s.com Sat Feb 18 02:47:55 2006 From: kemlo <@t> f2s.com (kemlo) Date: Sat Feb 18 02:48:05 2006 Subject: [Histonet] (no subject) In-Reply-To: Message-ID: <200602180847.k1I8lrRu025296@outmail.freedom2surf.net> Use .wav files under Windows stored on your main Server; give your Secretaries access to the sub directory and with some clever filing, Bingo. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mitchell, Jeannette M. Sent: 17 February 2006 17:32 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Hello Folks, We're looking for a digital dictation system for our grossing area to replace our dictaphones and tapes. We use CoPath for our pathology information system. Any suggestions ? thanks ! Jeannette Mitchell FAHC Burlington,VT Confidentiality Notice: This message, and any attachments, may contain information that is confidential, privileged, and/or protected from disclosure under state and federal laws that deal with the privacy and security of medical information. If you received this message in error or through inappropriate means, please reply to this message to notify the Sender that the message was received by you in error, and then permanently delete this message from all storage media, without forwarding or retaining a copy. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Sun Feb 19 09:09:22 2006 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Sun Feb 19 09:09:41 2006 Subject: [Histonet] P-cadherin Message-ID: Is anyone doing IHC for P-cadherin on formalin-fixed, paraffin-embedded tissue? If so, where do you get your antibody? Thanks! Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax From Adesupod <@t> aol.com Sun Feb 19 10:33:12 2006 From: Adesupod <@t> aol.com (Adesupod@aol.com) Date: Sun Feb 19 10:33:24 2006 Subject: [Histonet] Helicobacter Pylori Message-ID: <27.3d00fc2.3129f7c8@aol.com> Hi, Pls I want you histonetters to tell me the best silver technique for the demonstration of the Helicobacter Pylori. You guys are the best. Adesupo. From Rcartun <@t> harthosp.org Sun Feb 19 11:05:22 2006 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Sun Feb 19 11:06:18 2006 Subject: [Histonet] AMACR (P504S) Message-ID: I agree with Bob Richmond's comments. Our high molecular weight cytokeratin (clone 34BetaE12) works beautifully for demonstrating an absence of basal cells around malignant glands. We use P504S (from DAKO) on rare cases. Their antibody (clone 13H4) is labeled "IVD". I know that many labs are running multiple IP stains (HMW-CK, p63, P504S, and others) on prostate biopsies. To me, this is completely unnecessary for the majority of complicated cases where a diagnosis cannot be established on H&E stains, and is one reason why CMS is proposing MUEs to limit the number of IP stains that can be performed on a patient's specimen. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> 02/14/06 11:24AM >>> I am researching AMACR (P504s) and need some help. Who out there in histo-land is using it? Is this a Research Use Only antibody? Is a disclaimer in the report required? Any input would be appreciated...... Thanks, Michele Marggi Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tamustuff <@t> yahoo.com Sun Feb 19 14:44:45 2006 From: tamustuff <@t> yahoo.com (Stephen Sperry) Date: Sun Feb 19 14:44:52 2006 Subject: [Histonet] carcinoma vs. adenocarcinoma Message-ID: <20060219204445.22986.qmail@web37511.mail.mud.yahoo.com> Can anyone explain to me in layman terms (I'm taking undergrad histology) how to differentiate between a carcinoma and an adenocarcinoma? Thanks! Stephen --------------------------------- Yahoo! Mail Use Photomail to share photos without annoying attachments. From bhewlett <@t> cogeco.ca Sun Feb 19 14:59:21 2006 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Sun Feb 19 14:59:32 2006 Subject: [Histonet] carcinoma vs. adenocarcinoma References: <20060219204445.22986.qmail@web37511.mail.mud.yahoo.com> Message-ID: <000301c63597$5bb85a20$6400a8c0@mainbox> Steven, A carcinoma is a malignancy derived from epithelial cells (of any type), whereas an adenocarcinoma is derived from glandular or ductal epithelial cells. Bryan ----- Original Message ----- From: "Stephen Sperry" To: Sent: Sunday, February 19, 2006 3:44 PM Subject: [Histonet] carcinoma vs. adenocarcinoma > Can anyone explain to me in layman terms (I'm taking undergrad histology) > how to differentiate between a carcinoma and an adenocarcinoma? > > Thanks! > Stephen > > > --------------------------------- > Yahoo! Mail > Use Photomail to share photos without annoying attachments. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From ana.merino-trigo <@t> wanadoo.fr Sun Feb 19 15:47:12 2006 From: ana.merino-trigo <@t> wanadoo.fr (ana.merino-trigo) Date: Sun Feb 19 15:47:24 2006 Subject: [Histonet] Microarray References: Message-ID: <00bc01c6359e$0aa6b440$5b790a0a@BABA> Hi Lin, When I began to work with TMA I did have the same problem than you. I got great advices from the designer of the technology, Juha Kononen and also from Stephen Hewitt responsible of the TARP laboratory at the NCI. Basically the best results that I got were following this method: To build the block: -Use low melting point paraffin for the recipient block -Prior to Construction faced off the recipient block (in this way you will minimize the amount of realignment) To cut the block: Sectioning by "TEMPER" the blok 1.Once you finish the array place the block - FACE UP- in a oven at 35-37?C overnight 2.Next AM put it on a cold plate (face up) for an hour 3.Then reheat at 35-37?C for an hour 4.Then chill again, in the cold plate for an hour 5.Repeat the short cycle a few times. In this way to help to attach properly the spots to the surrounding paraffin, which avoids to lose spots during the microtome cutting. I cut the all block in the same day to avoid loosing slides during the realignment. I used 0.6 mm needles in the automated tissue arrayer ATA-27 from Beecher. By using this method I got a really good yield when using human xenografts and cell lines to build my TMA/CMA blocks. It's worth to try it, it made a big difference in my results. Hope this helps, Ana ----- Original Message ----- From: Thom Jensen To: Linresearch@aol.com ; histonet@lists.utsouthwestern.edu Sent: Friday, February 17, 2006 2:47 PM Subject: RE: [Histonet] Microarray Hey Lin, Which arrayer are you using? If you are using the Chemicon arrayer, their recipient needle is slightly bigger then the donor needle cores. You will need to set the depth of each recipient hole to match the donor core. If you are using the Beecher Instrument, then the donor core will fit snuggly in the Recipient hole unless you made the hole larger then the original hole on accident. Which often happens when people are learning to punch. If you are loosing cores from the array block while sectioning the array block on a microtome, then the problem is you haven't set the punches well. Put the finished array block in an oven on a glass slide (punch surface down on slide) at around 37 to 40 degrees Celsius for 15 minutes then press the glass slide and block together. This smashes the paraffin around the cores so the cores stick to the paraffin in the array block. If you don't do this step before you sections the array block the punches will fall out. This step is for both array instruments. I hope this helps, Cheers, Thom Jensen For more information of array instruction visit: www.arrayworkshop.com > >Hi, >Are there any hints to share on getting all the cores to stay evenly in the >recipient block and to get sections without losing cores? >Lin >_______________________________________________ >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 Free Edition. Version: 7.1.375 / Virus Database: 267.15.11/264 - Release Date: 17/02/2006 From ana.merino-trigo <@t> wanadoo.fr Sun Feb 19 15:49:39 2006 From: ana.merino-trigo <@t> wanadoo.fr (ana.merino-trigo) Date: Sun Feb 19 15:49:51 2006 Subject: [Histonet] Microarray Message-ID: <00cf01c6359e$627c5c60$5b790a0a@BABA> Hi Lin, When I began to work with TMA I did have the same problem than you. I got great advices from the designer of the technology, Juha Kononen and also from Stephen Hewitt responsible of the TARP laboratory at the NCI. Basically the best results that I got were following this method: To build the block: -Use low melting point paraffin for the recipient block -Prior to Construction faced off the recipient block (in this way you will minimize the amount of realignment) To cut the block: Sectioning by "TEMPER" the blok 1.Once you finish the array place the block - FACE UP- in a oven at 35-37?C overnight 2.Next AM put it on a cold plate (face up) for an hour 3.Then reheat at 35-37?C for an hour 4.Then chill again, in the cold plate for an hour 5.Repeat the short cycle a few times. In this way to help to attach properly the spots to the surrounding paraffin, which avoids to lose spots during the microtome cutting. I cut the all block in the same day to avoid loosing slides during the realignment. I used 0.6 mm needles in the automated tissue arrayer ATA-27 from Beecher. By using this method I got a really good yield when using human xenografts and cell lines to build my TMA/CMA blocks. It's worth to try it, it made a big difference in my results. Hope this helps, Ana ----- Original Message ----- From: Thom Jensen To: Linresearch@aol.com ; histonet@lists.utsouthwestern.edu Sent: Friday, February 17, 2006 2:47 PM Subject: RE: [Histonet] Microarray Hey Lin, Which arrayer are you using? If you are using the Chemicon arrayer, their recipient needle is slightly bigger then the donor needle cores. You will need to set the depth of each recipient hole to match the donor core. If you are using the Beecher Instrument, then the donor core will fit snuggly in the Recipient hole unless you made the hole larger then the original hole on accident. Which often happens when people are learning to punch. If you are loosing cores from the array block while sectioning the array block on a microtome, then the problem is you haven't set the punches well. Put the finished array block in an oven on a glass slide (punch surface down on slide) at around 37 to 40 degrees Celsius for 15 minutes then press the glass slide and block together. This smashes the paraffin around the cores so the cores stick to the paraffin in the array block. If you don't do this step before you sections the array block the punches will fall out. This step is for both array instruments. I hope this helps, Cheers, Thom Jensen For more information of array instruction visit: www.arrayworkshop.com > >Hi, >Are there any hints to share on getting all the cores to stay evenly in the >recipient block and to get sections without losing cores? >Lin >_______________________________________________ >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 Free Edition. Version: 7.1.375 / Virus Database: 267.15.11/264 - Release Date: 17/02/2006 From kemlo.rogerson <@t> waht.swest.nhs.uk Mon Feb 20 02:37:40 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Mon Feb 20 02:37:56 2006 Subject: [Histonet] carcinoma vs. adenocarcinoma Message-ID: Cytologicaly? Well an adenocarcinoma is a carcinoma of glandular cells; usually, but not always, illustrated by cells with a malignant nucleus eccentrically placed. The cytoplasm is whispy and may be vacuolated (not always) and the cells may be arranged in three dimensional clumps (not always); you may or may not notice a 'glandular' arrangement to the clump and there maybe a lack of polarity to the nuclear direction (they are pointing in different directions), there may be variation in nuclear size (anisonucleosis) with hyperchromasia (dark staining chromatin) or not. A carcinoma (if you are thinking of a squamous lesion) has usually a centrally placed malignant nucleus with 'hard' cytoplasm (sometimes keratinised, sometimes not). The cells have a tendency to be in 'sheets' but not always; anisonucleosis and hyperchromasia may also be exhibited or not. Depends how well differentiated the tumour is I suppose as a very poorly differentiated adenocarcinoma may look very similar to a very poorly differentiated carcinoma (squamous). Undifferentiated adeno and squamous carcinomas can look identical, or not! This is a very simplistic explaination....... IMHO as a mere Cytologist. If the above were true all the time, then Cytology would be easy most of the time. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Stephen Sperry [mailto:tamustuff@yahoo.com] Sent: Sunday, February 19, 2006 8:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] carcinoma vs. adenocarcinoma Can anyone explain to me in layman terms (I'm taking undergrad histology) how to differentiate between a carcinoma and an adenocarcinoma? Thanks! Stephen --------------------------------- Yahoo! Mail Use Photomail to share photos without annoying attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From andromeda_tm <@t> libero.it Mon Feb 20 04:58:36 2006 From: andromeda_tm <@t> libero.it (Massimo) Date: Mon Feb 20 04:58:47 2006 Subject: [Histonet] Ehrlich Biondi Heidenhain stain Message-ID: Hi all, does someone know the protocol for staining with the ?Ehrlich Biondi Heidenhain? method? Particularly how long has the specimen to stay in the staining solution? I have noticed, in previous, tests that is quit difficult to dye the nucleus with the methyl green contained in the solution. I am processing thin sections of mouse liver included in wax and fixed with Bouin liquid. I also wonder why the sections are crunched after cutting. There is a solution at this problem? Thanks in advance. Best Regards, Massimo Universit? degli Studi di Pisa Facolt? di Ingegneria Chimica Italy From abright <@t> brightinstruments.com Mon Feb 20 05:07:32 2006 From: abright <@t> brightinstruments.com (Alan Bright) Date: Mon Feb 20 04:58:49 2006 Subject: [Histonet] whole brain in paraffin Message-ID: Dear Nancy, Please feel free to contact me as I have experience with this microtome. One thing to note is that with sectioning large samples the design caused advancing on the return stroke which damaged the surface for the following sections. We had to redesign this fault on the microtomes we fitted into our cryostats. Best Regards Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: Nancy Lemke [mailto:nsnwl@neuro.hfh.edu] Sent: 16 February 2006 13:25 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] whole brain in paraffin Good Morning Histonet, I would like some information about the preparation and sectioning of 1-2cm slabs of human brain. The cutting will be done on an old Reichert Tetrander (sliding) microtome with a giant steel blade. If anyone who has experience with this sort of sectioning would email me I would love to ask some questions. Thank you in advance, Nancy Lemke Research Coordinator Hermelin Brain Tumor Center Henry Ford Hospital Detroit ======================================================================== ====== Go to http://henryford.com We're Henry Ford. We Can. 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The inner ear cells are fixed with formalin 10% and this is the soft tissue we are interested in. Our problem is that the bone surrounding the inner ear is not homogeneous. So, while some points are decalcified after 16 hours other need much more time. Then the inner ear cells are in contact with the RDO much before the sample is completely decalcified. I would like to ask if someone know how is the reaction we can expect from the cells fixed in formalin in contact with RDO. I'm wondering what you can advise me to do in our case, when the bone is not homogeneous, but it has to be decalicified. Thank you very much for your help. Best regards from, Maria Morell Maria Morell Laboratori d'Aplicacions Bioac?stiques Centre Tecnol?gic de Vilanova i la Geltr? Universitat Polit?cnica de Catalunya Avda. Rambla Exposici? s/n 08800-Vilanova i la Geltr? (Barcelona)- Spain Telf: 0034 938967227 http://www.lab.upc.edu From kemlo.rogerson <@t> waht.swest.nhs.uk Mon Feb 20 08:06:54 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Mon Feb 20 08:07:13 2006 Subject: [Histonet] RDO question Message-ID: So why not use EDTA? That can decalcify very dense bone including teeth. Try 5 to 10% at pH 6.5 to 7.5 to prevent protein hydrolysis. You could try 1M Trichloroacetic acid, nitric acid, formic acid, Picric acid OR EDTA at raised temperatures (26, 29, 30 or 40 degrees C) with alternating current (50 mA, 100 mA, 200 mA or 400 mA). With EDTA at 4oo mA the temperature will be 55 degrees C and you will get about 80% more calcium removed than at room temperature. Personally I think it's the raised temperature rather than the alternating current anyway, but I'm told temperature on its own is inconsistent. It's in Lillie's bible page 794. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: maria.morell@lab.upc.edu [mailto:maria.morell@lab.upc.edu] Sent: Monday, February 20, 2006 1:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RDO question Good morning Histonet! We are using RDO (a rapid decalcifier with hydrochloric acid) in our laboratory to decalcify very high density ear bones that surround the inner ear of marine mammals. The inner ear cells are fixed with formalin 10% and this is the soft tissue we are interested in. Our problem is that the bone surrounding the inner ear is not homogeneous. So, while some points are decalcified after 16 hours other need much more time. Then the inner ear cells are in contact with the RDO much before the sample is completely decalcified. I would like to ask if someone know how is the reaction we can expect from the cells fixed in formalin in contact with RDO. I'm wondering what you can advise me to do in our case, when the bone is not homogeneous, but it has to be decalicified. Thank you very much for your help. Best regards from, Maria Morell Maria Morell Laboratori d'Aplicacions Bioac?stiques Centre Tecnol?gic de Vilanova i la Geltr? Universitat Polit?cnica de Catalunya Avda. Rambla Exposici? s/n 08800-Vilanova i la Geltr? (Barcelona)- Spain Telf: 0034 938967227 http://www.lab.upc.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GDawson <@t> dynacaremilwaukee.com Mon Feb 20 11:08:07 2006 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Mon Feb 20 11:11:40 2006 Subject: [Histonet] P-cadherin Message-ID: Richard, I use the P-Cadherin from BD Transduction Laboratories (Cat.# 610228) 877-232-8995. Works pretty well on FFPE tissues as well. I believe I used it @ 1:100 dilution, pH 6.0 Citrate HIER, Monoclonal Envision+ Polymer (DAKO) 2ndary antibody detection. This was for a research project about a month ago but I think this is accurate. Good Luck, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI From thoeben <@t> bvderm.com Mon Feb 20 11:20:36 2006 From: thoeben <@t> bvderm.com (Tabatha Hoeben) Date: Mon Feb 20 11:20:29 2006 Subject: [Histonet] (no subject) Message-ID: <001201c63641$f7003d40$0c01a8c0@bvd.local> We are in need of a mohs tech for tomorrow. Any one available Tabatha Hoeben Boulder Valley Center For Dermatology thoeben@bvderm.com 303-604-1444 Fax 303-666-0911 From bwhitaker <@t> brownpathology.com Mon Feb 20 13:04:24 2006 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Mon Feb 20 13:04:30 2006 Subject: [Histonet] voice recognition software Message-ID: <000601c63650$76f466d0$3601a8c0@brownpathology.net> Hi Everyone, A friend asked me to inquire about voice recognition software. Is anyone out there using it? How well does it perform? Does anyone have any experience with Dragon that they would be willing to share? Thanks! Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 From jkiernan <@t> uwo.ca Mon Feb 20 13:25:07 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Mon Feb 20 13:25:16 2006 Subject: [Histonet] 20 micron sections Cresyl Violet Acetate stained References: <632ae8e2e68de84580523eede7cb1009@penguin.vmresearch.org> Message-ID: <43FA1793.165FD322@uwo.ca> There were 3 reasons for using 20um sections (though 6um sections were also used in the paper that you quote). 1. The spinal cords of interest were from people who had died of ALS, and motor neurons were not numerous. A thick section has more cells in it than a thin one.(The control subjects had plenty of motor neurons, of course, but the sections had to be the same thickness for quantitative comparisons.) 2. 20um was the thickest that was easily handled with paraffin sections. For neuroanatomical work, frozen sections are commonly cut at 40-80um for Nissl staining. 3. You need to include as much neuron as possible in the section, to show the shape and size. We did not have trouble with sections coming off the slides, which were coated (subbed) with chrome-gelatin. There was no difficulty with either the myelin stain (iron-eriochrome cyanine R; Page's method) or with the Nissl counterstain. Neutral red was used because it contrasted well with the blue myelin. If you're not interested in myelin you can use any cationic dye as a Nissl stain. Cresyl violet can be quite a fiddle; make sure you have a certified batch of the dye. Neutral red or toluidine blue (0.5%, pH 3.5 to 4) is easier to use. H&E is not suitable for staining neurons. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Patrick Laurie wrote: > > Hi Histonet, > > My group is interested in doing an anterior horn alpha motor neuron > count of ALS pts vs control. There is considerable literature about > these techniques, which has been most helpful, but the devil is in the > details. > > Most papers use 20um sections of the ventral horn stained with cresyl > violet acetate, or iron-chromoxane cyanine R method for myelin > (counterstained with .5% neutral red) (Dr. Kiernan's 1991 paper), or one > of many other methods for staining. I have 3 questions. First of all, > a technical question, how are 20um sections kept on the slides during > staining? I have done some experimenting, and found it very difficult > to keep them on. > > Secondly, how do the thick sections stain? The cresyl violet using will > not differentiate very well (using the regressive version), and the H&E > is too dark, I have yet to use any other method. > > And lastly, why 20um? Conventional neural histology seems to be done > thicker, but usually not that thick. What would be the benefit for > having sections that thick? > > Thanks Histonet, > > Patrick Laurie, HT (ASCP) > Neurogenomics Laboratory > Benaroya Research Institute > 1201 9th Ave > Seattle, Wa 98101 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DOOLEEO <@t> shands.ufl.edu Mon Feb 20 13:51:34 2006 From: DOOLEEO <@t> shands.ufl.edu (Elaine Dooley) Date: Mon Feb 20 13:52:16 2006 Subject: [Histonet] Oct-3/4 Message-ID: Dear Histonetters, I have been having some trouble trying to get this antibody to stain seminomas. It is Oct-3/4 a goat polyclonal antibody from Santa Cruz Biotechnology. We also purchased their mouse anti-goat biotin conjugated secondary. Any quick hints on making this work. I have tried it on the Ventana Benchmark (with changing the secondary antibody). I have tried overnight incubations and staining entirely by hand. I have tried Ventana's CC1 and also tried Cell Marque's trilogy for epitope retrial but have not had any of the desired nuclear staining the antibody is supposed to show. Any helpful hints would be greatly appreciated. Elaine Dooley HTL Shands Teaching Hospital Gainesville FL 352-265-0111 ext 7-2117 From sjchtascp <@t> yahoo.com Mon Feb 20 14:25:12 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Mon Feb 20 14:25:21 2006 Subject: [Histonet] sucrose sectioning Message-ID: <20060220202512.39971.qmail@web90210.mail.scd.yahoo.com> I work with a scientist that never seems pleased with my cryosectioning of fixed sucrose infiltrated liver. I have "tons" of experiance especially in clinical, and Mohs with cryosectioning and am trying to fine-tune my technique to research. I have reviewed, reviewed and reviewed material until the information is giving me a headach. Currectly I trim, fine trim on one section of blade the go to a fresh section to obtain my final section that I pick up. I would greatly appreciate any advice from those with experiance sectioning these tissues. Thanks you all, Steve --------------------------------- Yahoo! Mail Use Photomail to share photos without annoying attachments. From RCHIOVETTI <@t> aol.com Mon Feb 20 15:15:25 2006 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Mon Feb 20 15:16:07 2006 Subject: [Histonet] sucrose sectioning Message-ID: <26f.60d2e9f.312b8b6d@aol.com> In a message dated 2/20/2006 1:26:21 PM US Mountain Standard Time, sjchtascp@yahoo.com writes: > I work with a scientist that never seems pleased with my cryosectioning of > fixed sucrose infiltrated liver. Hi Steve, I'm most familiar with using sucrose as a cryoprotectant / infiltration medium when doing cryoultramicrotomy and immunolabeling at the EM level. In those cases, sucrose is normally used at around 2.3M concentration after light aldehyde fixation, and it usually cuts well at around -90 C (the cryosectioning kits for ultramicrotomes use liquid nitrogen, and they allow you to cut at very low temps). I never could get sucrose to cut well above about -60 to -65 C. It just turned to mush and became a sticky, stringy mess. A normal cryostat w/ independent specimen cooling should get down to about -50 C, but I'd think you'd be right on the edge of being able to use sucrose. Unless there's some reason not to use the stuff, maybe you could try O.C.T. or a similar cryo medium designed to be firm at higher temps (say round -20 to -30 C). Good luck! Cheers, Bob Robert (Bob) Chiovetti, Ph.D. The Microscope Works Arizona's Microscopy Resource 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA From lharris <@t> samhealth.org Mon Feb 20 16:03:32 2006 From: lharris <@t> samhealth.org (lharris@samhealth.org) Date: Mon Feb 20 16:03:51 2006 Subject: [Histonet] RE: AMACR (P504S) (Lori Harris) Message-ID: <5B3B26C4B71D5E469892D1860ABE10EA7F1E98@SHSEXVS01.int.samhealth.net> Richard, I found your response very interesting but could you explain to me what CMS & MUE stand for? Thanks! Lori A. Harris, HT (ASCP) Histology Section Leader GSRMC - Pathology 3600 NW Samaritan Drive Corvallis, OR 97330 1-541-768-6078 lharris@samhealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Sunday, February 19, 2006 10:02 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 27, Issue 29 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-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@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. P-cadherin (Richard Cartun) 2. Helicobacter Pylori (Adesupod@aol.com) 3. Re: AMACR (P504S) (Richard Cartun) ---------------------------------------------------------------------- Message: 1 Date: Sun, 19 Feb 2006 10:09:22 -0500 From: "Richard Cartun" Subject: [Histonet] P-cadherin To: Message-ID: Content-Type: text/plain; charset=US-ASCII Is anyone doing IHC for P-cadherin on formalin-fixed, paraffin-embedded tissue? If so, where do you get your antibody? Thanks! Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ------------------------------ Message: 2 Date: Sun, 19 Feb 2006 11:33:12 EST From: Adesupod@aol.com Subject: [Histonet] Helicobacter Pylori To: histonet@lists.utsouthwestern.edu Message-ID: <27.3d00fc2.3129f7c8@aol.com> Content-Type: text/plain; charset="US-ASCII" Hi, Pls I want you histonetters to tell me the best silver technique for the demonstration of the Helicobacter Pylori. You guys are the best. Adesupo. ------------------------------ Message: 3 Date: Sun, 19 Feb 2006 12:05:22 -0500 From: "Richard Cartun" Subject: Re: [Histonet] AMACR (P504S) To: , Message-ID: Content-Type: text/plain; charset=US-ASCII I agree with Bob Richmond's comments. Our high molecular weight cytokeratin (clone 34BetaE12) works beautifully for demonstrating an absence of basal cells around malignant glands. We use P504S (from DAKO) on rare cases. Their antibody (clone 13H4) is labeled "IVD". I know that many labs are running multiple IP stains (HMW-CK, p63, P504S, and others) on prostate biopsies. To me, this is completely unnecessary for the majority of complicated cases where a diagnosis cannot be established on H&E stains, and is one reason why CMS is proposing MUEs to limit the number of IP stains that can be performed on a patient's specimen. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> 02/14/06 11:24AM >>> I am researching AMACR (P504s) and need some help. Who out there in histo-land is using it? Is this a Research Use Only antibody? Is a disclaimer in the report required? Any input would be appreciated...... Thanks, Michele Marggi Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ 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 End of Histonet Digest, Vol 27, Issue 29 **************************************** Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From cforster <@t> umn.edu Mon Feb 20 16:28:38 2006 From: cforster <@t> umn.edu (cforster) Date: Mon Feb 20 16:29:55 2006 Subject: [Histonet] CD52 IHC Message-ID: <200602202228.k1KMScpN019762@saturn.software.umn.edu> Hello Histonetters, Is anyone doing teh CD52 on PPFE tissues? If so, would you share your vendor and protocol? I have tried several different things and cannot seem to get good staining. Thanks in advance. Colleen Forster HT(ASCP)QIHC Department of Laboratory Medicine and Pathology B173 PWB, MMC76 516 Delaware St. SE Minneapolis, MN 55455 612-626-1930 612-626-4660(f) From RCHIOVETTI <@t> aol.com Mon Feb 20 17:53:22 2006 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Mon Feb 20 17:53:35 2006 Subject: [Histonet] Re: Sucrose Sectioning Message-ID: <193.5190ec55.312bb072@aol.com> Here's a suggestion from Andi Grantham, University of Arizona Health Sciences Center: ********************************************************************* Sometimes before sectioning something that has been infiltrated with sucrose I pop it into OCT and allow it to sit there for a while to let the OCT try to replace some of the sucrose. Then turn the specimen holder down real low to cut. Sometimes this works, sometimes not. Andi ********************************************************************* Robert (Bob) Chiovetti, Ph.D. The Microscope Works Arizona's Microscopy Resource 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA From mlafrini <@t> csmlab.com Tue Feb 21 06:38:38 2006 From: mlafrini <@t> csmlab.com (Michael LaFriniere) Date: Tue Feb 21 06:39:53 2006 Subject: [Histonet] voice recognition software Message-ID: Bonnie, I implemented Dragon with the Co Path system last month, so far we are demonstrating excellent acceptance with the grossing area as well as the Pathologists. It takes a little time to get all to buy into using the system, however, the productivity cost decrease, ( primarily due to reduction in transcription cost) far exceeds the "up front" time and cost involved to implement..... Good luck, Michael Michael R. LaFriniere Executive Director Cytology Services of Maryland (CSM) 301-206-2555 ext 27 301-206-2595 fax michael.lafriniere@csmlab.com >>> "Bonnie Whitaker" 2/20/2006 2:04 pm >>> Hi Everyone, A friend asked me to inquire about voice recognition software. Is anyone out there using it? How well does it perform? Does anyone have any experience with Dragon that they would be willing to share? Thanks! Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------- From pbrask <@t> comcast.net Tue Feb 21 07:28:28 2006 From: pbrask <@t> comcast.net (Peggy Brask) Date: Tue Feb 21 07:28:34 2006 Subject: [Histonet] Do you love your job? Message-ID: I am a second semester student at Argosy University in the HT program. I was wondering what the pros and cons are in the Histonet world. I have been in the service industry in one form or another for the last 30 years. This is a total diversion from my past life and I would like to know what you love about your job and what you hate about your job. If anyone would like to give me some input, I would greatly appreciate it. Thanks, Peggy From clcses <@t> gmail.com Tue Feb 21 07:50:04 2006 From: clcses <@t> gmail.com (Carmen Contreras-Sesvold) Date: Tue Feb 21 07:50:13 2006 Subject: [Histonet] re: Oct-3/4 staining of seminomas Message-ID: <46a3be380602210550i73ee0edct9136ef747c7265d7@mail.gmail.com> What is your positive control? Check your secondary to verify if it is working. If everything else is working it might be your primary. Call Santa Cruz and ask to speak to their tech service for that primary to see if they can assist. You can also ask them to provide another lot number that for the primary (for free). They have surperb tech support. I have had the best service from them. Carmen From JWEEMS <@t> sjha.org Tue Feb 21 08:30:16 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Tue Feb 21 08:30:26 2006 Subject: [Histonet] FW: ASCP Action Alert: High Priority Action Alert!! MUE Proposal Could Undermine Quality Care, Devastate Clinical Laboratories Message-ID: <83AACDB0810528418AA106F9AE9B7F7E01305850@sjhaexc02.sjha.org> I hesitated before I send this, but we in the US are facing a pathology crisis if we don't do something. For those not in the US I apologize, the rest of us need to do what we can to help! Thanks, j Joyce Weems Pathology Manager Saint Joseph?s Hospital of Atlanta 404-851-7376 404-851-7831 - fax High Priority Action Alert!! MUE Proposal Could Undermine Quality Care, Devastate Clinical Laboratories! America?s clinical laboratories are under assault by a federal initiative and ASCP needs your help to defeat it! The Centers for Medicare and Medicaid (CMS) recently proposed a massive CPT coding edit, placing stringent caps on the units of service that can be provided patients for pathology and laboratory services as well as other medical services. The initiative could have profound consequences for patient care and could also undermine the financial viability of numerous clinical laboratories. One clinical laboratory estimates that the proposal could reduce its Medicare Part A reimbursement by 35 percent and its Part B reimbursement by 10 percent. This proposal might be one of the most harmful policy initiatives faced by clinical laboratories in years. What?s the Proposal? CMS?s MUE (Medically Unbelievable Edits) proposal sets limits on the number of units of services that can be billed per patient per day. The MUE proposal sets limits on almost 1200 pathology and laboratory CPT Codes. According to CMS, the coding edits proposed prevent billing for items that are either (1) anatomically impossible (can?t remove more than one appendix) or (2) medically unreasonable (more than one pacemaker). Unfortunately, many of the coding edits proposed by CMS are inconsistent with existing guidelines for diagnosing and treating patients. Under the proposal CPT code 88305 may not be reimbursed at more than two units of service per patient per day while 88342 would be capped at 4. MUEs being proposed would allow contractors to ?automatically deny the services without stopping the claim for routine or complex review, even if documentation is attached.? What is ASCP Doing? ASCP has written CMS Administrator Mark McClellan to protest the MUE proposal, stating that the coding edits are flawed and threaten quality patient care. In addition, ASCP is launching a multi-pronged effort to fight this initiative. This initiative will reach out to ASCP members to seek their technical input on these flaws and to generate grassroots opposition to the CMS proposal. ASCP plans to compile this information to provide further evidence that CMS? MUE proposal is fundamentally flawed. What Can I Do to Help? To bolster the case that the MUE proposal is flawed, ASCP needs its members? technical input to help identify clinical scenarios where the MUE proposal is inconsistent with the proper practice of laboratory medicine. Journal articles or practice guidelines recommending a greater unit of service than that allowed under the MUE proposal would be particularly helpful. ASCP urges its members and others in the laboratory community to access ASCP?s e-Advocacy Center to review a condensed version of the MUE proposal. This condensed list focuses on the top 100 pathology and laboratory codes by reimbursement. While it does not cover all 1200 CPT codes, the e-Advocacy Center includes instructions for obtaining a copy of the entire MUE proposal. Also on the e-Advocacy Center is a template for providing technical input to ASCP. A copy of ASCP?s letter to CMS Administrator Mark McClellan is also on the e-Advocacy Center. The following are two examples of the type of input that helps bolster the case that the MUE proposal is flawed. For CPT code 88305 (Level IV ? surgical pathology, gross and microscopic examination), the MUE proposal limits reimbursement to only 2 procedures per site per day. Guidelines developed by the American Gastroenterology Association (AGA) and American Cancer Society (ACS) recommend that for patients with ulcerative colitis, ?two to four random biopsy specimens should be obtained every 10 cm from the entire colon for a total of approximately 40 biopsies per patient per colonoscopy, and additional samples should be obtained at any suspicious area.? (7th Symposium on Inflammatory Bowel Diseases and Salicylates). For CPT code 88342 (immunocytochemistry (w/tissue immunoperoxidase), each antibody), the MUE proposed limit is 4 units of service per patient per day. Some tumors cannot be ascertained on histologic grounds, however, and require the aid of immunohistochemical stains. In the case of anaplastic tumors, a panel of seven immunohistochemical stains is needed (Gatter and Mason, Seminars in Oncology, Vol. 9, No. 4, 1982). Moreover, in an article by Raab (Arch Pathol Lab Med, Vol. 124, Aug. 2000), the author concluded that ?immunohistochemistry is extremely cost-effective,? with a more favorable cost-benefit ratio than other medical procedures. To provide input on the CMS MUE proposal, please use e-Advocacy Center to request the necessary documents from the ASCP Washington Office. We?ll send you a copy of ASCP?s letter to CMS along with a copy of the MUE proposal affecting the top 100 pathology and laboratory service codes (by reimbursement). If you?d like a copy of the MUE proposal affecting every pathology and laboratory code, please indicate that you would like the expanded document as well. ASCP thanks you for your efforts to help us better serve you and the laboratory community! Tell a friend: Not everyone receives ASCP's Action Alerts, so please forward this message to your peers and co-workers! ASCP America's Health Depends on Its Laboratories ? 2003 American Society for Clinical Pathology 33 West Monroe Street, Suite 1600, Chicago, IL 60603 312.541.4999 ~ info@ascp.org ABOUT THIS MESSAGE You are receiving this email because you are a member of ASCP, have attended ASCP programs/meetings or made a purchase from ASCP. If you no longer wish to receive emails of this kind from ASCP, please login to change your email preferences at ascp.org. This email message complies with all CAN-SPAM 2004 regulations. It was sent to you by the AMERICAN SOCIETY FOR CLINICAL PATHOLOGY, 33 West Monroe, Suite 1600, Chicago, IL 60603 Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From tkngflght <@t> yahoo.com Tue Feb 21 09:00:53 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Tue Feb 21 09:01:03 2006 Subject: [Histonet] A call to action for US AP/Histology professionals In-Reply-To: <83AACDB0810528418AA106F9AE9B7F7E01305850@sjhaexc02.sjha.org> Message-ID: <20060221150053.71471.qmail@web50907.mail.yahoo.com> Hello Folks-- There is another issue we should consider a call to paper the Senate with letters stating our opinions....The AFIP faces closure due to realignment by base closures (BRAC)--this includes the pathology center AND the school!! We've lost so many good schools, and the AFIP is among the best. There is a bill to relocate core services of the AFIP to another center and the link below provides an easy way to write you your senator in support of this bill: http://capwiz.com/ascpath/issues/alert/?alertid=8171231&type=CO Please take a minute to show your support for our science on MUE billing (Peggy's email of earlier today),AFIP closure, and two additional issues threatening continued viability of lab science in this country. The form is easy to fill out, and you can forward the link to several friends once you submit your letter (it will give you another pop-up for sharing the letter once you click to submit yours) to increase the impact of our unified voice. This link will lead you to both letters and you can send it by email or snail mail: http://capwiz.com/ascpath/home/ Thank you for your continued interest and support tp maintain visibility and viability of our craft. Together we DO make a difference. Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one Tech at a time. 281.883.7704 c 281.852.9457 o admin@fullstaff.org Sign up for the FREE monthly newsletter AP News--updates, tricks of the trade and current issues for Anatomic Pathology Clinical Labs. Send a 'subscribe' request to APNews@fullstaff.org. Please include your name and specialty in the body of the email. From Sue.Kapoor <@t> uhsi.org Tue Feb 21 09:49:39 2006 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Tue Feb 21 09:48:53 2006 Subject: [Histonet] StarFrost Plus Slides for IHC Message-ID: Hi everyone, I'm considering changing from using SuperFrost Plus slides to StarFrost Plus slides from Mercedes Medical for my immunos but I'd hate to start getting unreliable staining...is anyone currently using StarFrost Plus slides for immunos?? Thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From tfranzod <@t> dal.ca Tue Feb 21 10:00:26 2006 From: tfranzod <@t> dal.ca (Tamara Franz-Odendaal) Date: Tue Feb 21 10:02:25 2006 Subject: [Histonet] immunohistochemistry fish tissue - help Message-ID: <07a701c636ff$f4c6d6b0$6d20ad81@tam> I have two questions: 1) Does anyone have a good protocol for immunohistochemistry working with fish tissue embedded in paraffin wax? 2) Any tricks with working with collagen antibodies (apart from hyaluronidase treatment)? I have been trying without success to do the above, I have tried an Alexa 488 secondary as well as a horse radish peroxidase secondary. Neither are working and I am wondering if it is something peculiar to fish tissue. What are good blocks to use to prevent autoimmunofluorescence, or what secondary do you suggest. I am getting high backgrounds due to secondary. And also no signal in the area of interest (cartilage) so the primaries are also not working well. I am starting to wonder if the antibodies are at fault but before I order more wanted to know if anyone has any good working protocols. I am working on larval tissue not embryonic. zebrafish. Or do you suggest whole mount immunohistochemistry? Thanks Tamara Dr. Tamara Franz-Odendaal Dalhousie University Halifax, Canada From rjbuesa <@t> yahoo.com Tue Feb 21 10:21:10 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Feb 21 10:21:19 2006 Subject: [Histonet] Do you love your job? In-Reply-To: Message-ID: <20060221162110.98958.qmail@web61224.mail.yahoo.com> Hi Peggy: What others love or hate about a job has no individual relevance at all. You will have to experience that feeling by yourself. If you have dexterity, if you like to cook, if you like to knit, if you like to innovate and experiment, if you have a good smell sense, if you enjoy colours, if you look at what you manually have done and feel proud about it, if you understand the importance of doing things to help others, if you don't mind working more than required just to make sure that what you are doing is finished correctly, if you like at least some of the aforesaid, you will like histotechnology, if not, you are in a wrong switch from your previous activities. At least, those are some of the things I love; there are more, and what I don't like mostly are things related with human nature, and those you will find and hate in any profession. Hope this will give you an idea! ren? J. Peggy Brask wrote: I am a second semester student at Argosy University in the HT program. I was wondering what the pros and cons are in the Histonet world. I have been in the service industry in one form or another for the last 30 years. This is a total diversion from my past life and I would like to know what you love about your job and what you hate about your job. If anyone would like to give me some input, I would greatly appreciate it. Thanks, Peggy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! From dgaupp <@t> tulane.edu Tue Feb 21 11:04:41 2006 From: dgaupp <@t> tulane.edu (dgaupp@tulane.edu) Date: Tue Feb 21 11:04:46 2006 Subject: [Histonet] wright-giemsa stain on tissue Message-ID: <1140541481.43fb4829095a0@webmail.tulane.edu> Histonet: Has anyone ever heard of staining wright-giemsa stain on paraffin embedded tissue(rat bone)? I've stained in the past blood smears and bone marrow smears. Just wondering if anyone out there in histoland has ever done so. I checked out histonet archives and did not find any results of paraffin processed tissue stained with wright-giemsa. Thanks, Dina From rjbuesa <@t> yahoo.com Tue Feb 21 11:16:46 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Feb 21 11:16:54 2006 Subject: [Histonet] wright-giemsa stain on tissue In-Reply-To: <1140541481.43fb4829095a0@webmail.tulane.edu> Message-ID: <20060221171646.97141.qmail@web61214.mail.yahoo.com> Hi Dina: I am attaching a paper I published on the subject to your e-mail address. I hope this will help you. Ren? J. dgaupp@tulane.edu wrote: Histonet: Has anyone ever heard of staining wright-giemsa stain on paraffin embedded tissue(rat bone)? I've stained in the past blood smears and bone marrow smears. Just wondering if anyone out there in histoland has ever done so. I checked out histonet archives and did not find any results of paraffin processed tissue stained with wright-giemsa. Thanks, Dina _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Mail Use Photomail to share photos without annoying attachments. From Jackie.O'Connor <@t> abbott.com Tue Feb 21 11:53:23 2006 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Tue Feb 21 11:53:52 2006 Subject: [Histonet] Do you love your job? Message-ID: Perhaps Peggy is looking for a different spin on this type of work - many people aren't suited for histology. If you can't stand the sight or smell of blood - you're not suited. I have a sister who is an ER nurse - she doesn't mind debriding head wounds on people, yet to see a hysterectomy sample on my dissecting board brought her to dry heaves - - the only thing that ever really grossed me out was when I found 1/2 a shoe in my lab refer over a long weekend - of course it also contained 1/2 a foot. My eldest daughter has a BS in biology, but could never work with animal tissues - it makes her cry. In my opinion, the worst part about histology is the potential for chemical and biological exposure - yeah, you can take safeguards, but the risk is still there - let's face it, the odds of the general public being exposed to formaldehyde went away when they quit putting it in Mr. Bubble bubble bath - not to mention silver nitrate, chloroform, osmium tetroxide - the list goes on. Other things that have bothered me over the last 35 years are fetal samples - but I'm a Mom. Sometimes you have to not think about the patient behind the sample - it can get too depressing - on the other hand, I've learned more about anatomy and the process of disease that I would have ever learned anywhere else. I think the key is asking a lot of questions if you have a pathologist who is willing to explain - I've been fortunate working with many pathologists who were terrific teachers. I like my job - I'm proud of what I do. I'm at the point in my life where I'm helping to find a cure for cancer instead of just seeing cancer specimens - and that's way cool. It's a great and honorable profession, and someone has to do it. I'm glad to be a part of it. Jackie O' Rene J Buesa Sent by: histonet-bounces@lists.utsouthwestern.edu 02/21/2006 10:21 AM To: pbrask@comcast.net, histonet@lists.utsouthwestern.edu cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: Re: [Histonet] Do you love your job? Hi Peggy: What others love or hate about a job has no individual relevance at all. You will have to experience that feeling by yourself. If you have dexterity, if you like to cook, if you like to knit, if you like to innovate and experiment, if you have a good smell sense, if you enjoy colours, if you look at what you manually have done and feel proud about it, if you understand the importance of doing things to help others, if you don't mind working more than required just to make sure that what you are doing is finished correctly, if you like at least some of the aforesaid, you will like histotechnology, if not, you are in a wrong switch from your previous activities. At least, those are some of the things I love; there are more, and what I don't like mostly are things related with human nature, and those you will find and hate in any profession. Hope this will give you an idea! ren? J. Peggy Brask wrote: I am a second semester student at Argosy University in the HT program. I was wondering what the pros and cons are in the Histonet world. I have been in the service industry in one form or another for the last 30 years. This is a total diversion from my past life and I would like to know what you love about your job and what you hate about your job. If anyone would like to give me some input, I would greatly appreciate it. Thanks, Peggy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TMcNemar <@t> lmhealth.org Tue Feb 21 12:00:25 2006 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Tue Feb 21 12:01:00 2006 Subject: [Histonet] StarFrost Plus Slides for IHC Message-ID: <61FA27BFA9398E4CA9184FF9B0759B08660D47@nt_exchange.lmhealth.org> We routinly use them without a problem. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Kapoor, Sue [mailto:Sue.Kapoor@uhsi.org] Sent: Tuesday, February 21, 2006 10:50 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] StarFrost Plus Slides for IHC Hi everyone, I'm considering changing from using SuperFrost Plus slides to StarFrost Plus slides from Mercedes Medical for my immunos but I'd hate to start getting unreliable staining...is anyone currently using StarFrost Plus slides for immunos?? Thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kathy.Johnston <@t> CLS.ab.ca Tue Feb 21 12:19:19 2006 From: Kathy.Johnston <@t> CLS.ab.ca (Kathy.Johnston@CLS.ab.ca) Date: Tue Feb 21 12:19:28 2006 Subject: [Histonet] wright-giemsa stain on tissue Message-ID: <4BC300747AF87A48BCDF8E48BC2885CE013652C2@mail1.calgary.com> We do this for some of our research pathologists, and I would have to say that it seems to work quite well. If you need a protocol, let me know! Kathy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of dgaupp@tulane.edu Sent: February 21, 2006 10:05 AM To: Histonet Subject: [Histonet] wright-giemsa stain on tissue Histonet: Has anyone ever heard of staining wright-giemsa stain on paraffin embedded tissue(rat bone)? I've stained in the past blood smears and bone marrow smears. Just wondering if anyone out there in histoland has ever done so. I checked out histonet archives and did not find any results of paraffin processed tissue stained with wright-giemsa. Thanks, Dina _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify the sender immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. From Sue.Kapoor <@t> uhsi.org Tue Feb 21 12:21:01 2006 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Tue Feb 21 12:20:18 2006 Subject: [Histonet] Do you love your job? Message-ID: I what I find difficult is when animal histo/research folks talk about perfusion and sacrificing. I know you're doing great and wonderful things but it would be too hard for me. Other than that I've been amazed by some of the things I've seen. And I've always been extremely proud to be part of this field. The biggest thing I HATE is techs that loose sight of that and it shows in their work (or lack of). Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jackie M O'Connor Sent: Tuesday, February 21, 2006 11:53 AM To: Rene J Buesa Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Do you love your job? Perhaps Peggy is looking for a different spin on this type of work - many people aren't suited for histology. If you can't stand the sight or smell of blood - you're not suited. I have a sister who is an ER nurse - she doesn't mind debriding head wounds on people, yet to see a hysterectomy sample on my dissecting board brought her to dry heaves - - the only thing that ever really grossed me out was when I found 1/2 a shoe in my lab refer over a long weekend - of course it also contained 1/2 a foot. My eldest daughter has a BS in biology, but could never work with animal tissues - it makes her cry. In my opinion, the worst part about histology is the potential for chemical and biological exposure - yeah, you can take safeguards, but the risk is still there - let's face it, the odds of the general public being exposed to formaldehyde went away when they quit putting it in Mr. Bubble bubble bath - not to mention silver nitrate, chloroform, osmium tetroxide - the list goes on. Other things that have bothered me over the last 35 years are fetal samples - but I'm a Mom. Sometimes you have to not think about the patient behind the sample - it can get too depressing - on the other hand, I've learned more about anatomy and the process of disease that I would have ever learned anywhere else. I think the key is asking a lot of questions if you have a pathologist who is willing to explain - I've been fortunate working with many pathologists who were terrific teachers. I like my job - I'm proud of what I do. I'm at the point in my life where I'm helping to find a cure for cancer instead of just seeing cancer specimens - and that's way cool. It's a great and honorable profession, and someone has to do it. I'm glad to be a part of it. Jackie O' Rene J Buesa Sent by: histonet-bounces@lists.utsouthwestern.edu 02/21/2006 10:21 AM From brittakarpe <@t> googlemail.com Tue Feb 21 13:22:40 2006 From: brittakarpe <@t> googlemail.com (Britta Karpe) Date: Tue Feb 21 13:22:52 2006 Subject: [Histonet] ICH Mouse tissue (kidney, vessels) Message-ID: <65a51e7a0602211122m4863fbaay@mail.gmail.com> Hello Histonetters! I?m a newbie in IHC and look for staining-methods of CD 40 / CD 154 / CRP / Nitrotyrosin that work on (SNX)-Mouse kidney and vessels. I tried an CRP polyclonal Antibody from Sigma with different unmasking methods (heat/enzymes) and the ABC/AEC Detection System. I found no specific staining but a lot of background which i tried to block with Normal Serum and some Protein-Blocks (without success). The Sigma Datasheet indicates that the antibody is specific for human, they have not tested it on mouse tissues yet. My colleague tried the same with CD40 and CD 154 without success. Some papers show the results of CRP-stains on human kidney but the pictures aren?t very convincing. Somebody told me that only the Podocytes show a staining with CRP, and the other stained cells (mesangial, tubular proximal cells..) are only background... is that true? Thanks for all advices! Britta From lesley.bechtold <@t> jax.org Tue Feb 21 14:23:06 2006 From: lesley.bechtold <@t> jax.org (Lesley Bechtold) Date: Tue Feb 21 14:24:47 2006 Subject: [Histonet] Seeking Information Message-ID: <20060221152306519.00000002804@spikey> Hi Histonetters, Our administration is asking us to provide them with bench-marking information that demonstrates where we are relative to everyone else as far as the Histological Services we are offering our scientists and pathologists. As funding gets tighter and tighter, we want to be sure that we are providing the best and most cost-effective service we can. I am hoping that many of you out there are looking for similar information and will be willing to participate. To make this as painless as possible, we have developed a short, ten- question survey. I have put in a link (see below) that will take you to our survey page. Once there, simply click on "Histology" and answer the survey questions as they pertain to your Histology Lab. Ten questions as well as some institutional information is all that is required. Anyone who participates will receive a copy of all the results, minus any personal information. I hope you can use this information as well. Thank you in advance! Lesley Bechtold http://www.jax.org/static/sci_survey/sci_survey_index.html Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 From anh2006 <@t> med.cornell.edu Tue Feb 21 14:55:56 2006 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Tue Feb 21 14:56:06 2006 Subject: [Histonet] Donkey serum Message-ID: Where is everyone buying their donkey serum from these days? Thanks! -- From sjchtascp <@t> yahoo.com Tue Feb 21 14:59:11 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Tue Feb 21 14:59:20 2006 Subject: [Histonet] histology jobs- Message-ID: <20060221205911.29054.qmail@web90201.mail.scd.yahoo.com> looking for several good sites to look for histology positions, any type, FT, PT, contract ets. --------------------------------- Yahoo! Autos. Looking for a sweet ride? Get pricing, reviews, & more on new and used cars. From lao_ji <@t> yahoo.com Tue Feb 21 16:24:54 2006 From: lao_ji <@t> yahoo.com (Jimmy Lao) Date: Tue Feb 21 16:25:04 2006 Subject: [Histonet] Donkey serum In-Reply-To: Message-ID: <20060221222454.63901.qmail@web30705.mail.mud.yahoo.com> >From Jackson Lab, Cat#017-000-121. Hope this help! Jimmy --- "Andrea T. Hooper" wrote: > Where is everyone buying their donkey serum from > these days? > > Thanks! > -- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From failm <@t> musc.edu Tue Feb 21 16:26:26 2006 From: failm <@t> musc.edu (Mildred Fail) Date: Tue Feb 21 16:26:56 2006 Subject: [Histonet] Do you love your job? Message-ID: Hi Peggy, I used to love my job, I would be excited about coming to work. Looking at a well cut well stained slide was like looking at a fine painting. It didn't matter whether I had worked on the slide or not. I liked the beauty of it. Wonderful colors. Working out a problematic antibody gave me a sense of accomplishment and pride. I loved reading and learning more about the stains going back in time to learn how it was done before dyes were so readily available. Though I am fortunate that most of the pathologists focus on the forward progress much more emphasis is placed by others on the negative. One of my techs says it is a thankless job. Seldom does anyone walk in to say how good a stain looks and most of our stains are quite good, picture perfect. But good stains are expected so therefore not always appreciated. You have to have a real passion for histology to ignore all the negative, to keep on trying to perfect your craft, to keep on learning. it is the passion for histology that allows you to see a problematic stain as a challenge. Once your focus is off the slides, the patient , the stains, you loose your footing and the job is no longer exciting. Rena Fail >>> "Peggy Brask" 02/21/06 08:28AM >>> I am a second semester student at Argosy University in the HT program. I was wondering what the pros and cons are in the Histonet world. I have been in the service industry in one form or another for the last 30 years. This is a total diversion from my past life and I would like to know what you love about your job and what you hate about your job. If anyone would like to give me some input, I would greatly appreciate it. Thanks, Peggy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Tue Feb 21 17:42:07 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Tue Feb 21 17:42:15 2006 Subject: [Histonet] Help for Transcription Message-ID: <83AACDB0810528418AA106F9AE9B7F7E01305872@sjhaexc02.sjha.org> I need to justify additional transcriptionist hours. Would you please do me a favor and ask your transcription department for the number of minutes they have transcribed for Feb to date? - This can be found in the transcription system, I assume. We have Lanier and we have access to that info. I'd like to know # of cases Number of gross minutes dictated: Number of micro minutes dictated: Number of transcriptionists: Thanks for your help! Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From JWEEMS <@t> sjha.org Tue Feb 21 17:47:33 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Tue Feb 21 17:47:41 2006 Subject: [Histonet] Help for Transcription Message-ID: <83AACDB0810528418AA106F9AE9B7F7E01305873@sjhaexc02.sjha.org> Someone brought to my attention last week that all my posts to Histonet have an attachment. I'd never paid attention as I didn't read them! Does anyone know what I can do to stop them? It may be something in my hospital network. I apologize until I can find out what is wrong. j Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Weems, Joyce Sent: Tuesday, February 21, 2006 6:42 PM To: Histonet Subject: [Histonet] Help for Transcription I need to justify additional transcriptionist hours. Would you please do me a favor and ask your transcription department for the number of minutes they have transcribed for Feb to date? - This can be found in the transcription system, I assume. We have Lanier and we have access to that info. I'd like to know # of cases Number of gross minutes dictated: Number of micro minutes dictated: Number of transcriptionists: Thanks for your help! Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From lpwenk <@t> sbcglobal.net Tue Feb 21 18:32:43 2006 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Tue Feb 21 18:33:23 2006 Subject: [Histonet] PGP 9.5 Message-ID: <001b01c63747$7ede4760$ede49440@HPPav2> Anyone have experience with PGP 9.5 (Protein Gene Product)? It demonstrates nerve fibers. One of our pathologists wants to use it on skin biopsies for diagnosis of neuropathies. We would prefer peroxidase for paraffin sections, rather than FITC or FS. Just wondering if people are having better luck with monoclonals or polyclonals, and if there is a difference in animal species for the primary (we'll be doing this on human skin). Also - all the papers we find mention thick sections and using a confocal microscope. Anyone doing PGP on 5 um sections and/or with a regular light microscope? One of my students is doing this as her research project. She's done a lot of reading on it, and most papers don't discuss the actual procedure or the type of antibody, etc. They are on the research aspect, or the diagnostic aspect, not on what histotechs need to do the procedures. Those articles that do mention techniques have very different methods (of course), and are missing important information needed by histotechs. We would appreciate a couple of little hints of what works for "real life histotechs", to get us started in the right direction for use in a routine clinical immunohistochemistry lab. Feel free to contact me directly. Vendors included. I'll summarize and relay the info back to Histonet. Thanks in advance. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 From tkngflght <@t> yahoo.com Tue Feb 21 18:42:13 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Tue Feb 21 18:42:21 2006 Subject: [Histonet] Do you love your job? In-Reply-To: Message-ID: <20060222004213.2308.qmail@web50910.mail.yahoo.com> I've been fortunate to spend more of my career in places where the Doc would come in singing because he loved our quality of work and had helped to make his job a song instead of complaining about the one slide that had a wrinkle. I wish more of us were in places where our work--which we take pride in and spend more time at than we do with our families--was audibly appreciated daily. Please trust that these places still exist---I talk with my old co-workers who still create a place of 'second family' in the lab and I work with them every day as an agent. It IS about the patient. When I get stuck in the mire (we all do), I volunteer on the cancer floor at the hospital where I work. To give a backrub to the patient who's bone marrow I cut the day before, or walk the hall with the lymphoma patient we worked up last week who will qualify for the new therapy because of our work...of course they don't know that I am the one who took part in their diagnosis, but it sure keeps my focus on the patient, not the people who focus on the negative....the ones who can't see the forest for the trees. Just my two cents. Cheryl Mildred Fail wrote: Hi Peggy, I used to love my job, I would be excited about coming to work. Looking at a well cut well stained slide was like looking at a fine painting. It didn't matter whether I had worked on the slide or not. I liked the beauty of it. Wonderful colors. Working out a problematic antibody gave me a sense of accomplishment and pride. I loved reading and learning more about the stains going back in time to learn how it was done before dyes were so readily available. Though I am fortunate that most of the pathologists focus on the forward progress much more emphasis is placed by others on the negative. One of my techs says it is a thankless job. Seldom does anyone walk in to say how good a stain looks and most of our stains are quite good, picture perfect. But good stains are expected so therefore not always appreciated. You have to have a real passion for histology to ignore all the negative, to keep on trying to perfect your craft, to keep on learning. it is the passion for histology that allows you to see a problematic stain as a challenge. Once your focus is off the slides, the patient , the stains, you loose your footing and the job is no longer exciting. Rena Fail >>> "Peggy Brask" 02/21/06 08:28AM >>> I am a second semester student at Argosy University in the HT program. I was wondering what the pros and cons are in the Histonet world. I have been in the service industry in one form or another for the last 30 years. This is a total diversion from my past life and I would like to know what you love about your job and what you hate about your job. If anyone would like to give me some input, I would greatly appreciate it. Thanks, Peggy _______________________________________________ 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 From jhnspam <@t> aol.com Tue Feb 21 20:58:20 2006 From: jhnspam <@t> aol.com (jhnspam@aol.com) Date: Tue Feb 21 20:58:48 2006 Subject: [Histonet] Tissue Processor Information Message-ID: <154.60c0dc9f.312d2d4c@aol.com> Does anyone out there have any information on the TBS Tissue Processor? What are you likes and dislikes? What about their service? Thanks, Jennifer From Megan.Clarke <@t> hnehealth.nsw.gov.au Tue Feb 21 23:58:26 2006 From: Megan.Clarke <@t> hnehealth.nsw.gov.au (Megan Clarke) Date: Wed Feb 22 00:00:26 2006 Subject: [Histonet] antibody information Message-ID: Hi histonetters We are interested in purchasing the following antibodies: Desmoplakin (1 and 2) - From Progen Biotech, Heidelberg Germany. D2-40 - a marker for mesotheliomas and epithelial cells. not sure of source of antibody. Cav 3 ( Caveolin-3) Monoclonal antiody from Transduction laboratories, Lexiton, KY Firstly, if anyone is using any of these antibodies could you please furnish me with the agent/s (if any are located in Australia) that supplies these antibodies and secondly if you have used them are they "good little antibodies"? Thank you in anticipation Zenobia Haffajee IHC Dept HAPS Newcastle Australia. From ian.montgomery <@t> bio.gla.ac.uk Wed Feb 22 03:37:15 2006 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Wed Feb 22 03:37:27 2006 Subject: Fw: [Histonet] PGP 9.5 Message-ID: <005801c63793$90d5f8e0$532ed182@ibls.gla.ac.uk> Peggy, PGP9.5 (monoclonal) from Novocastra works a treat. Whole mounts, cryostat, FFPE, with any demonstration technique works. Used it on various species, including man, with success. Ian. ----- Original Message ----- From: "Lee & Peggy Wenk" To: Sent: Wednesday, February 22, 2006 12:32 AM Subject: [Histonet] PGP 9.5 > Anyone have experience with PGP 9.5 (Protein Gene Product)? It demonstrates > nerve fibers. One of our pathologists wants to use it on skin biopsies for > diagnosis of neuropathies. > > We would prefer peroxidase for paraffin sections, rather than FITC or FS. > Just wondering if people are having better luck with monoclonals or > polyclonals, and if there is a difference in animal species for the primary > (we'll be doing this on human skin). > > Also - all the papers we find mention thick sections and using a confocal > microscope. Anyone doing PGP on 5 um sections and/or with a regular light > microscope? > > One of my students is doing this as her research project. She's done a lot > of reading on it, and most papers don't discuss the actual procedure or the > type of antibody, etc. They are on the research aspect, or the diagnostic > aspect, not on what histotechs need to do the procedures. Those articles > that do mention techniques have very different methods (of course), and are > missing important information needed by histotechs. > > We would appreciate a couple of little hints of what works for "real life > histotechs", to get us started in the right direction for use in a routine > clinical immunohistochemistry lab. > > Feel free to contact me directly. Vendors included. I'll summarize and relay > the info back to Histonet. > > Thanks in advance. > > Peggy A. Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From ian.montgomery <@t> bio.gla.ac.uk Wed Feb 22 04:18:12 2006 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Wed Feb 22 04:18:24 2006 Subject: Fw: [Histonet] Do you love your job? Message-ID: <006d01c63799$4981b8c0$532ed182@ibls.gla.ac.uk> Peggy, Been a Histologist for 39 years and 6 months what do I think? Against, low pay and long hours. For, like others, the sheer joy of looking at stained sections. If, just once in your career, you prepare a section and see structures that are of textbook quality that's enough. For me, a loop of Henle, the perfect example stained with Masson. Receptors in the skin stained with silver, Meissner, Krause, Golgi, ring and whorl and free endings at the stratum basale. A modified Mallory demonstrating the cells in the anterior pituitary. Better still, having in your collection slides that were prepared in the 1800's and still works of extreme beauty. Plus, the joy of lecturing. Standing in a lecture theatre or classroom and waxing lyrically on a subject you enjoy and hoping that this enthusiasm reaches some of the students. Then pride at the end when students tell you they really enjoyed the lecture and that they now have a clearer understanding of the topic. No doubt your new career will have its peaks and troughs but if you start in a forward thinking lab with good technicians then the love of the subject will remain. Ian. >>> "Peggy Brask" 02/21/06 08:28AM >>> I am a second semester student at Argosy University in the HT program. I was wondering what the pros and cons are in the Histonet world. I have been in the service industry in one form or another for the last 30 years. This is a total diversion from my past life and I would like to know what you love about your job and what you hate about your job. If anyone would like to give me some input, I would greatly appreciate it. Thanks, Peggy _______________________________________________ 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 From Jeannette.Mitchell <@t> vtmednet.org Wed Feb 22 05:31:12 2006 From: Jeannette.Mitchell <@t> vtmednet.org (Mitchell, Jeannette M.) Date: Wed Feb 22 05:31:23 2006 Subject: [Histonet] microwaves Message-ID: Hi Folks, What is everyone using when doing special stains for a microwave. Is there any OSHA regulations for microwaves??? Thanks for you help. Jeannette Mitchell Burlington, Vt Confidentiality Notice: This message, and any attachments, may contain information that is confidential, privileged, and/or protected from disclosure under state and federal laws that deal with the privacy and security of medical information. If you received this message in error or through inappropriate means, please reply to this message to notify the Sender that the message was received by you in error, and then permanently delete this message from all storage media, without forwarding or retaining a copy. From Jeannette.Mitchell <@t> vtmednet.org Wed Feb 22 05:33:21 2006 From: Jeannette.Mitchell <@t> vtmednet.org (Mitchell, Jeannette M.) Date: Wed Feb 22 05:33:30 2006 Subject: [Histonet] (no subject) Message-ID: Hi Folks, What is everyone using when doing special stains for a microwave. Is there any OSHA regulations for microwaves??? Thanks for you help. Jeannette Mitchell Burlington Confidentiality Notice: This message, and any attachments, may contain information that is confidential, privileged, and/or protected from disclosure under state and federal laws that deal with the privacy and security of medical information. If you received this message in error or through inappropriate means, please reply to this message to notify the Sender that the message was received by you in error, and then permanently delete this message from all storage media, without forwarding or retaining a copy. From twebster <@t> nmcinc.org Wed Feb 22 06:25:43 2006 From: twebster <@t> nmcinc.org (Tim Webster) Date: Wed Feb 22 06:25:58 2006 Subject: [Histonet] RE: Histonet Digest, Vol 27, Issue 31 Message-ID: <0CBF5C420BD8AD4AAEDED774BD4A240F17B94C@exchange.nmcinc.org> Hi Bonnie I use "Dragon Naturally Speaking" quite a lot in my home office. There is a significant amount of time teaching it to recognize your voice, and especially if you use lots of odd :) medical words such as "Microcytosis" and "Menometrorrhagia", there is an even longer period. You will have to edit a lot at first. However- Once you get past that part, it writes pretty well. You also have to adjust your dictation style and not watch the screen for the words to appear. There is a lag time between you speaking and the software putting it into context and displaying the sentence, so if you watch the screen, the dictation is very slow and jumpy. If you write a lot, then give it a shot. If you don't, you probably won't use it. Good luck Tim Webster Histology Specialist Northwestern Medical Center 133 Fairfield Street St Albans, VT 05478 (802) 524-1070 x4349 (Dept. & Voice mail) twebster@nmcinc.org Statement of Confidentiality This message and any attachments are from NMC and intended only for the addressee(s). The information contained may include privileged or other confidential information. Unauthorized review, forwarding, printing, copying or distribution is strictly prohibited. Any opinions expressed are those of the author and not necessarily those of Norhtwestern Medical Center. If you should receive this message in error, please delete it and notify the sender. Thank you. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Tuesday, February 21, 2006 1:36 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 27, Issue 31 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-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@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. voice recognition software (Bonnie Whitaker) 2. Re: 20 micron sections Cresyl Violet Acetate stained (John A. Kiernan) 3. Oct-3/4 (Elaine Dooley) 4. sucrose sectioning (Steven Coakley) 5. Re: sucrose sectioning (RCHIOVETTI@aol.com) 6. RE: AMACR (P504S) (Lori Harris) (lharris@samhealth.org) 7. CD52 IHC (cforster) 8. Re: Sucrose Sectioning (RCHIOVETTI@aol.com) 9. Re: voice recognition software (Michael LaFriniere) 10. Do you love your job? (Peggy Brask) 11. re: Oct-3/4 staining of seminomas (Carmen Contreras-Sesvold) 12. FW: ASCP Action Alert: High Priority Action Alert!! MUE Proposal Could Undermine Quality Care, Devastate Clinical Laboratories (Weems, Joyce) 13. A call to action for US AP/Histology professionals (Cheryl) 14. StarFrost Plus Slides for IHC (Kapoor, Sue) 15. immunohistochemistry fish tissue - help (Tamara Franz-Odendaal) 16. Re: Do you love your job? (Rene J Buesa) 17. wright-giemsa stain on tissue (dgaupp@tulane.edu) 18. Re: wright-giemsa stain on tissue (Rene J Buesa) 19. Re: Do you love your job? (Jackie M O'Connor) ---------------------------------------------------------------------- Message: 1 Date: Mon, 20 Feb 2006 13:04:24 -0600 From: "Bonnie Whitaker" Subject: [Histonet] voice recognition software To: Message-ID: <000601c63650$76f466d0$3601a8c0@brownpathology.net> Content-Type: text/plain; charset="us-ascii" Hi Everyone, A friend asked me to inquire about voice recognition software. Is anyone out there using it? How well does it perform? Does anyone have any experience with Dragon that they would be willing to share? Thanks! Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 ------------------------------ Message: 2 Date: Mon, 20 Feb 2006 14:25:07 -0500 From: "John A. Kiernan" Subject: Re: [Histonet] 20 micron sections Cresyl Violet Acetate stained To: Patrick Laurie Cc: "histonet@lists.utsouthwestern.edu" Message-ID: <43FA1793.165FD322@uwo.ca> Content-Type: text/plain; charset=us-ascii There were 3 reasons for using 20um sections (though 6um sections were also used in the paper that you quote). 1. The spinal cords of interest were from people who had died of ALS, and motor neurons were not numerous. A thick section has more cells in it than a thin one.(The control subjects had plenty of motor neurons, of course, but the sections had to be the same thickness for quantitative comparisons.) 2. 20um was the thickest that was easily handled with paraffin sections. For neuroanatomical work, frozen sections are commonly cut at 40-80um for Nissl staining. 3. You need to include as much neuron as possible in the section, to show the shape and size. We did not have trouble with sections coming off the slides, which were coated (subbed) with chrome-gelatin. There was no difficulty with either the myelin stain (iron-eriochrome cyanine R; Page's method) or with the Nissl counterstain. Neutral red was used because it contrasted well with the blue myelin. If you're not interested in myelin you can use any cationic dye as a Nissl stain. Cresyl violet can be quite a fiddle; make sure you have a certified batch of the dye. Neutral red or toluidine blue (0.5%, pH 3.5 to 4) is easier to use. H&E is not suitable for staining neurons. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Patrick Laurie wrote: > > Hi Histonet, > > My group is interested in doing an anterior horn alpha motor neuron > count of ALS pts vs control. There is considerable literature about > these techniques, which has been most helpful, but the devil is in the > details. > > Most papers use 20um sections of the ventral horn stained with cresyl > violet acetate, or iron-chromoxane cyanine R method for myelin > (counterstained with .5% neutral red) (Dr. Kiernan's 1991 paper), or one > of many other methods for staining. I have 3 questions. First of all, > a technical question, how are 20um sections kept on the slides during > staining? I have done some experimenting, and found it very difficult > to keep them on. > > Secondly, how do the thick sections stain? The cresyl violet using will > not differentiate very well (using the regressive version), and the H&E > is too dark, I have yet to use any other method. > > And lastly, why 20um? Conventional neural histology seems to be done > thicker, but usually not that thick. What would be the benefit for > having sections that thick? > > Thanks Histonet, > > Patrick Laurie, HT (ASCP) > Neurogenomics Laboratory > Benaroya Research Institute > 1201 9th Ave > Seattle, Wa 98101 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Mon, 20 Feb 2006 14:51:34 -0500 From: "Elaine Dooley" Subject: [Histonet] Oct-3/4 To: Cc: Kenneth Iczkowski Message-ID: Content-Type: text/plain; charset=US-ASCII Dear Histonetters, I have been having some trouble trying to get this antibody to stain seminomas. It is Oct-3/4 a goat polyclonal antibody from Santa Cruz Biotechnology. We also purchased their mouse anti-goat biotin conjugated secondary. Any quick hints on making this work. I have tried it on the Ventana Benchmark (with changing the secondary antibody). I have tried overnight incubations and staining entirely by hand. I have tried Ventana's CC1 and also tried Cell Marque's trilogy for epitope retrial but have not had any of the desired nuclear staining the antibody is supposed to show. Any helpful hints would be greatly appreciated. Elaine Dooley HTL Shands Teaching Hospital Gainesville FL 352-265-0111 ext 7-2117 ------------------------------ Message: 4 Date: Mon, 20 Feb 2006 12:25:12 -0800 (PST) From: Steven Coakley Subject: [Histonet] sucrose sectioning To: Histonet@lists.utsouthwestern.edu Message-ID: <20060220202512.39971.qmail@web90210.mail.scd.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I work with a scientist that never seems pleased with my cryosectioning of fixed sucrose infiltrated liver. I have "tons" of experiance especially in clinical, and Mohs with cryosectioning and am trying to fine-tune my technique to research. I have reviewed, reviewed and reviewed material until the information is giving me a headach. Currectly I trim, fine trim on one section of blade the go to a fresh section to obtain my final section that I pick up. I would greatly appreciate any advice from those with experiance sectioning these tissues. Thanks you all, Steve --------------------------------- Yahoo! Mail Use Photomail to share photos without annoying attachments. ------------------------------ Message: 5 Date: Mon, 20 Feb 2006 16:15:25 EST From: RCHIOVETTI@aol.com Subject: Re: [Histonet] sucrose sectioning To: sjchtascp@yahoo.com, Histonet@lists.utsouthwestern.edu Message-ID: <26f.60d2e9f.312b8b6d@aol.com> Content-Type: text/plain; charset="US-ASCII" In a message dated 2/20/2006 1:26:21 PM US Mountain Standard Time, sjchtascp@yahoo.com writes: > I work with a scientist that never seems pleased with my cryosectioning of > fixed sucrose infiltrated liver. Hi Steve, I'm most familiar with using sucrose as a cryoprotectant / infiltration medium when doing cryoultramicrotomy and immunolabeling at the EM level. In those cases, sucrose is normally used at around 2.3M concentration after light aldehyde fixation, and it usually cuts well at around -90 C (the cryosectioning kits for ultramicrotomes use liquid nitrogen, and they allow you to cut at very low temps). I never could get sucrose to cut well above about -60 to -65 C. It just turned to mush and became a sticky, stringy mess. A normal cryostat w/ independent specimen cooling should get down to about -50 C, but I'd think you'd be right on the edge of being able to use sucrose. Unless there's some reason not to use the stuff, maybe you could try O.C.T. or a similar cryo medium designed to be firm at higher temps (say round -20 to -30 C). Good luck! Cheers, Bob Robert (Bob) Chiovetti, Ph.D. The Microscope Works Arizona's Microscopy Resource 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA ------------------------------ Message: 6 Date: Mon, 20 Feb 2006 14:03:32 -0800 From: Subject: [Histonet] RE: AMACR (P504S) (Lori Harris) To: Message-ID: <5B3B26C4B71D5E469892D1860ABE10EA7F1E98@SHSEXVS01.int.samhealth.net> Content-Type: text/plain; charset="iso-8859-1" Richard, I found your response very interesting but could you explain to me what CMS & MUE stand for? Thanks! Lori A. Harris, HT (ASCP) Histology Section Leader GSRMC - Pathology 3600 NW Samaritan Drive Corvallis, OR 97330 1-541-768-6078 lharris@samhealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Sunday, February 19, 2006 10:02 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 27, Issue 29 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-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@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. P-cadherin (Richard Cartun) 2. Helicobacter Pylori (Adesupod@aol.com) 3. Re: AMACR (P504S) (Richard Cartun) ---------------------------------------------------------------------- Message: 1 Date: Sun, 19 Feb 2006 10:09:22 -0500 From: "Richard Cartun" Subject: [Histonet] P-cadherin To: Message-ID: Content-Type: text/plain; charset=US-ASCII Is anyone doing IHC for P-cadherin on formalin-fixed, paraffin-embedded tissue? If so, where do you get your antibody? Thanks! Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ------------------------------ Message: 2 Date: Sun, 19 Feb 2006 11:33:12 EST From: Adesupod@aol.com Subject: [Histonet] Helicobacter Pylori To: histonet@lists.utsouthwestern.edu Message-ID: <27.3d00fc2.3129f7c8@aol.com> Content-Type: text/plain; charset="US-ASCII" Hi, Pls I want you histonetters to tell me the best silver technique for the demonstration of the Helicobacter Pylori. You guys are the best. Adesupo. ------------------------------ Message: 3 Date: Sun, 19 Feb 2006 12:05:22 -0500 From: "Richard Cartun" Subject: Re: [Histonet] AMACR (P504S) To: , Message-ID: Content-Type: text/plain; charset=US-ASCII I agree with Bob Richmond's comments. Our high molecular weight cytokeratin (clone 34BetaE12) works beautifully for demonstrating an absence of basal cells around malignant glands. We use P504S (from DAKO) on rare cases. Their antibody (clone 13H4) is labeled "IVD". I know that many labs are running multiple IP stains (HMW-CK, p63, P504S, and others) on prostate biopsies. To me, this is completely unnecessary for the majority of complicated cases where a diagnosis cannot be established on H&E stains, and is one reason why CMS is proposing MUEs to limit the number of IP stains that can be performed on a patient's specimen. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> 02/14/06 11:24AM >>> I am researching AMACR (P504s) and need some help. Who out there in histo-land is using it? Is this a Research Use Only antibody? Is a disclaimer in the report required? Any input would be appreciated...... Thanks, Michele Marggi Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ 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 End of Histonet Digest, Vol 27, Issue 29 **************************************** Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ------------------------------ Message: 7 Date: Mon, 20 Feb 2006 16:28:38 CST From: cforster Subject: [Histonet] CD52 IHC To: histonet@lists.utsouthwestern.edu Message-ID: <200602202228.k1KMScpN019762@saturn.software.umn.edu> Content-Type: TEXT/plain; CHARSET=US-ASCII Hello Histonetters, Is anyone doing teh CD52 on PPFE tissues? If so, would you share your vendor and protocol? I have tried several different things and cannot seem to get good staining. Thanks in advance. Colleen Forster HT(ASCP)QIHC Department of Laboratory Medicine and Pathology B173 PWB, MMC76 516 Delaware St. SE Minneapolis, MN 55455 612-626-1930 612-626-4660(f) ------------------------------ Message: 8 Date: Mon, 20 Feb 2006 18:53:22 EST From: RCHIOVETTI@aol.com Subject: [Histonet] Re: Sucrose Sectioning To: histonet@lists.utsouthwestern.edu Message-ID: <193.5190ec55.312bb072@aol.com> Content-Type: text/plain; charset="US-ASCII" Here's a suggestion from Andi Grantham, University of Arizona Health Sciences Center: ********************************************************************* Sometimes before sectioning something that has been infiltrated with sucrose I pop it into OCT and allow it to sit there for a while to let the OCT try to replace some of the sucrose. Then turn the specimen holder down real low to cut. Sometimes this works, sometimes not. Andi ********************************************************************* Robert (Bob) Chiovetti, Ph.D. The Microscope Works Arizona's Microscopy Resource 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA ------------------------------ Message: 9 Date: Tue, 21 Feb 2006 07:38:38 -0500 From: "Michael LaFriniere" Subject: Re: [Histonet] voice recognition software To: "Bonnie Whitaker" , Message-ID: Content-Type: text/plain; charset="US-ASCII" Bonnie, I implemented Dragon with the Co Path system last month, so far we are demonstrating excellent acceptance with the grossing area as well as the Pathologists. It takes a little time to get all to buy into using the system, however, the productivity cost decrease, ( primarily due to reduction in transcription cost) far exceeds the "up front" time and cost involved to implement..... Good luck, Michael Michael R. LaFriniere Executive Director Cytology Services of Maryland (CSM) 301-206-2555 ext 27 301-206-2595 fax michael.lafriniere@csmlab.com >>> "Bonnie Whitaker" 2/20/2006 2:04 pm >>> Hi Everyone, A friend asked me to inquire about voice recognition software. Is anyone out there using it? How well does it perform? Does anyone have any experience with Dragon that they would be willing to share? Thanks! Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------- ------------------------------ Message: 10 Date: Tue, 21 Feb 2006 07:28:28 -0600 From: "Peggy Brask" Subject: [Histonet] Do you love your job? To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" I am a second semester student at Argosy University in the HT program. I was wondering what the pros and cons are in the Histonet world. I have been in the service industry in one form or another for the last 30 years. This is a total diversion from my past life and I would like to know what you love about your job and what you hate about your job. If anyone would like to give me some input, I would greatly appreciate it. Thanks, Peggy ------------------------------ Message: 11 Date: Tue, 21 Feb 2006 08:50:04 -0500 From: "Carmen Contreras-Sesvold" Subject: [Histonet] re: Oct-3/4 staining of seminomas To: histonetters Message-ID: <46a3be380602210550i73ee0edct9136ef747c7265d7@mail.gmail.com> Content-Type: text/plain; charset=ISO-8859-1 What is your positive control? Check your secondary to verify if it is working. If everything else is working it might be your primary. Call Santa Cruz and ask to speak to their tech service for that primary to see if they can assist. You can also ask them to provide another lot number that for the primary (for free). They have surperb tech support. I have had the best service from them. Carmen ------------------------------ Message: 12 Date: Tue, 21 Feb 2006 09:30:16 -0500 From: "Weems, Joyce" Subject: [Histonet] FW: ASCP Action Alert: High Priority Action Alert!! MUE Proposal Could Undermine Quality Care, Devastate Clinical Laboratories To: "Histonet" Message-ID: <83AACDB0810528418AA106F9AE9B7F7E01305850@sjhaexc02.sjha.org> Content-Type: text/plain; charset="utf-8" I hesitated before I send this, but we in the US are facing a pathology crisis if we don't do something. For those not in the US I apologize, the rest of us need to do what we can to help! Thanks, j Joyce Weems Pathology Manager Saint Joseph???s Hospital of Atlanta 404-851-7376 404-851-7831 - fax High Priority Action Alert!! MUE Proposal Could Undermine Quality Care, Devastate Clinical Laboratories! America???s clinical laboratories are under assault by a federal initiative and ASCP needs your help to defeat it! The Centers for Medicare and Medicaid (CMS) recently proposed a massive CPT coding edit, placing stringent caps on the units of service that can be provided patients for pathology and laboratory services as well as other medical services. The initiative could have profound consequences for patient care and could also undermine the financial viability of numerous clinical laboratories. One clinical laboratory estimates that the proposal could reduce its Medicare Part A reimbursement by 35 percent and its Part B reimbursement by 10 percent. This proposal might be one of the most harmful policy initiatives faced by clinical laboratories in years. What???s the Proposal? CMS???s MUE (Medically Unbelievable Edits) proposal sets limits on the number of units of services that can be billed per patient per day. The MUE proposal sets limits on almost 1200 pathology and laboratory CPT Codes. According to CMS, the coding edits proposed prevent billing for items that are either (1) anatomically impossible (can???t remove more than one appendix) or (2) medically unreasonable (more than one pacemaker). Unfortunately, many of the coding edits proposed by CMS are inconsistent with existing guidelines for diagnosing and treating patients. Under the proposal CPT code 88305 may not be reimbursed at more than two units of service per patient per day while 88342 would be capped at 4. MUEs being proposed would allow contractors to ???automatically deny the services without stopping the claim for routine or complex review, even if documentation is attached.??? What is ASCP Doing? ASCP has written CMS Administrator Mark McClellan to protest the MUE proposal, stating that the coding edits are flawed and threaten quality patient care. In addition, ASCP is launching a multi-pronged effort to fight this initiative. This initiative will reach out to ASCP members to seek their technical input on these flaws and to generate grassroots opposition to the CMS proposal. ASCP plans to compile this information to provide further evidence that CMS??? MUE proposal is fundamentally flawed. What Can I Do to Help? To bolster the case that the MUE proposal is flawed, ASCP needs its members?? ? technical input to help identify clinical scenarios where the MUE proposal is inconsistent with the proper practice of laboratory medicine. Journal articles or practice guidelines recommending a greater unit of service than that allowed under the MUE proposal would be particularly helpful. ASCP urges its members and others in the laboratory community to access ASCP???s e-Advocacy Center to review a condensed version of the MUE proposal. This condensed list focuses on the top 100 pathology and laboratory codes by reimbursement. While it does not cover all 1200 CPT codes, the e-Advocacy Center includes instructions for obtaining a copy of the entire MUE proposal. Also on the e-Advocacy Center is a template for providing technical input to ASCP. A copy of ASCP???s letter to CMS Administrator Mark McClellan is also on the e-Advocacy Center. The following are two examples of the type of input that helps bolster the case that the MUE proposal is flawed. For CPT code 88305 (Level IV ??? surgical pathology, gross and microscopic examination), the MUE proposal limits reimbursement to only 2 procedures per site per day. Guidelines developed by the American Gastroenterology Association (AGA) and American Cancer Society (ACS) recommend that for patients with ulcerative colitis, ???two to four random biopsy specimens should be obtained every 10 cm from the entire colon for a total of approximately 40 biopsies per patient per colonoscopy, and additional samples should be obtained at any suspicious area.??? (7th Symposium on Inflammatory Bowel Diseases and Salicylates). For CPT code 88342 (immunocytochemistry (w/tissue immunoperoxidase), each antibody), the MUE proposed limit is 4 units of service per patient per day. Some tumors cannot be ascertained on histologic grounds, however, and require the aid of immunohistochemical stains. In the case of anaplastic tumors, a panel of seven immunohistochemical stains is needed (Gatter and Mason, Seminars in Oncology, Vol. 9, No. 4, 1982). Moreover, in an article by Raab (Arch Pathol Lab Med, Vol. 124, Aug. 2000), the author concluded that ???immunohistochemistry is extremely cost-effective,??? with a more favorable cost-benefit ratio than other medical procedures. To provide input on the CMS MUE proposal, please use e-Advocacy Center to request the necessary documents from the ASCP Washington Office. We???ll send you a copy of ASCP???s letter to CMS along with a copy of the MUE proposal affecting the top 100 pathology and laboratory service codes (by reimbursement). If you???d like a copy of the MUE proposal affecting every pathology and laboratory code, please indicate that you would like the expanded document as well. ASCP thanks you for your efforts to help us better serve you and the laboratory community! Tell a friend: Not everyone receives ASCP's Action Alerts, so please forward this message to your peers and co-workers! ASCP America's Health Depends on Its Laboratories ?? 2003 American Society for Clinical Pathology 33 West Monroe Street, Suite 1600, Chicago, IL 60603 312.541.4999 ~ info@ascp.org ABOUT THIS MESSAGE You are receiving this email because you are a member of ASCP, have attended ASCP programs/meetings or made a purchase from ASCP. If you no longer wish to receive emails of this kind from ASCP, please login to change your email preferences at ascp.org. This email message complies with all CAN-SPAM 2004 regulations. It was sent to you by the AMERICAN SOCIETY FOR CLINICAL PATHOLOGY, 33 West Monroe, Suite 1600, Chicago, IL 60603 Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. ------------------------------ Message: 13 Date: Tue, 21 Feb 2006 07:00:53 -0800 (PST) From: Cheryl Subject: [Histonet] A call to action for US AP/Histology professionals To: Histonet Message-ID: <20060221150053.71471.qmail@web50907.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello Folks-- There is another issue we should consider a call to paper the Senate with letters stating our opinions....The AFIP faces closure due to realignment by base closures (BRAC)--this includes the pathology center AND the school!! We've lost so many good schools, and the AFIP is among the best. There is a bill to relocate core services of the AFIP to another center and the link below provides an easy way to write you your senator in support of this bill: http://capwiz.com/ascpath/issues/alert/?alertid=8171231&type=CO Please take a minute to show your support for our science on MUE billing (Peggy's email of earlier today),AFIP closure, and two additional issues threatening continued viability of lab science in this country. The form is easy to fill out, and you can forward the link to several friends once you submit your letter (it will give you another pop-up for sharing the letter once you click to submit yours) to increase the impact of our unified voice. This link will lead you to both letters and you can send it by email or snail mail: http://capwiz.com/ascpath/home/ Thank you for your continued interest and support tp maintain visibility and viability of our craft. Together we DO make a difference. Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one Tech at a time. 281.883.7704 c 281.852.9457 o admin@fullstaff.org Sign up for the FREE monthly newsletter AP News--updates, tricks of the trade and current issues for Anatomic Pathology Clinical Labs. Send a 'subscribe' request to APNews@fullstaff.org. Please include your name and specialty in the body of the email. ------------------------------ Message: 14 Date: Tue, 21 Feb 2006 09:49:39 -0600 From: "Kapoor, Sue" Subject: [Histonet] StarFrost Plus Slides for IHC To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=iso-8859-1 Hi everyone, I'm considering changing from using SuperFrost Plus slides to StarFrost Plus slides from Mercedes Medical for my immunos but I'd hate to start getting unreliable staining...is anyone currently using StarFrost Plus slides for immunos?? Thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 ------------------------------ Message: 15 Date: Tue, 21 Feb 2006 12:00:26 -0400 From: "Tamara Franz-Odendaal" Subject: [Histonet] immunohistochemistry fish tissue - help To: Message-ID: <07a701c636ff$f4c6d6b0$6d20ad81@tam> Content-Type: text/plain; charset="iso-8859-1" I have two questions: 1) Does anyone have a good protocol for immunohistochemistry working with fish tissue embedded in paraffin wax? 2) Any tricks with working with collagen antibodies (apart from hyaluronidase treatment)? I have been trying without success to do the above, I have tried an Alexa 488 secondary as well as a horse radish peroxidase secondary. Neither are working and I am wondering if it is something peculiar to fish tissue. What are good blocks to use to prevent autoimmunofluorescence, or what secondary do you suggest. I am getting high backgrounds due to secondary. And also no signal in the area of interest (cartilage) so the primaries are also not working well. I am starting to wonder if the antibodies are at fault but before I order more wanted to know if anyone has any good working protocols. I am working on larval tissue not embryonic. zebrafish. Or do you suggest whole mount immunohistochemistry? Thanks Tamara Dr. Tamara Franz-Odendaal Dalhousie University Halifax, Canada ------------------------------ Message: 16 Date: Tue, 21 Feb 2006 08:21:10 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Do you love your job? To: pbrask@comcast.net, histonet@lists.utsouthwestern.edu Message-ID: <20060221162110.98958.qmail@web61224.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi Peggy: What others love or hate about a job has no individual relevance at all. You will have to experience that feeling by yourself. If you have dexterity, if you like to cook, if you like to knit, if you like to innovate and experiment, if you have a good smell sense, if you enjoy colours, if you look at what you manually have done and feel proud about it, if you understand the importance of doing things to help others, if you don't mind working more than required just to make sure that what you are doing is finished correctly, if you like at least some of the aforesaid, you will like histotechnology, if not, you are in a wrong switch from your previous activities. At least, those are some of the things I love; there are more, and what I don't like mostly are things related with human nature, and those you will find and hate in any profession. Hope this will give you an idea! ren? J. Peggy Brask wrote: I am a second semester student at Argosy University in the HT program. I was wondering what the pros and cons are in the Histonet world. I have been in the service industry in one form or another for the last 30 years. This is a total diversion from my past life and I would like to know what you love about your job and what you hate about your job. If anyone would like to give me some input, I would greatly appreciate it. Thanks, Peggy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! ------------------------------ Message: 17 Date: Tue, 21 Feb 2006 11:04:41 -0600 From: dgaupp@tulane.edu Subject: [Histonet] wright-giemsa stain on tissue To: Histonet Message-ID: <1140541481.43fb4829095a0@webmail.tulane.edu> Content-Type: text/plain; charset=ISO-8859-1 Histonet: Has anyone ever heard of staining wright-giemsa stain on paraffin embedded tissue(rat bone)? I've stained in the past blood smears and bone marrow smears. Just wondering if anyone out there in histoland has ever done so. I checked out histonet archives and did not find any results of paraffin processed tissue stained with wright-giemsa. Thanks, Dina ------------------------------ Message: 18 Date: Tue, 21 Feb 2006 09:16:46 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] wright-giemsa stain on tissue To: dgaupp@tulane.edu, Histonet Message-ID: <20060221171646.97141.qmail@web61214.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi Dina: I am attaching a paper I published on the subject to your e-mail address. I hope this will help you. Ren? J. dgaupp@tulane.edu wrote: Histonet: Has anyone ever heard of staining wright-giemsa stain on paraffin embedded tissue(rat bone)? I've stained in the past blood smears and bone marrow smears. Just wondering if anyone out there in histoland has ever done so. I checked out histonet archives and did not find any results of paraffin processed tissue stained with wright-giemsa. Thanks, Dina _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Mail Use Photomail to share photos without annoying attachments. ------------------------------ Message: 19 Date: Tue, 21 Feb 2006 11:53:23 -0600 From: "Jackie M O'Connor" Subject: Re: [Histonet] Do you love your job? To: Rene J Buesa Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" Perhaps Peggy is looking for a different spin on this type of work - many people aren't suited for histology. If you can't stand the sight or smell of blood - you're not suited. I have a sister who is an ER nurse - she doesn't mind debriding head wounds on people, yet to see a hysterectomy sample on my dissecting board brought her to dry heaves - - the only thing that ever really grossed me out was when I found 1/2 a shoe in my lab refer over a long weekend - of course it also contained 1/2 a foot. My eldest daughter has a BS in biology, but could never work with animal tissues - it makes her cry. In my opinion, the worst part about histology is the potential for chemical and biological exposure - yeah, you can take safeguards, but the risk is still there - let's face it, the odds of the general public being exposed to formaldehyde went away when they quit putting it in Mr. Bubble bubble bath - not to mention silver nitrate, chloroform, osmium tetroxide - the list goes on. Other things that have bothered me over the last 35 years are fetal samples - but I'm a Mom. Sometimes you have to not think about the patient behind the sample - it can get too depressing - on the other hand, I've learned more about anatomy and the process of disease that I would have ever learned anywhere else. I think the key is asking a lot of questions if you have a pathologist who is willing to explain - I've been fortunate working with many pathologists who were terrific teachers. I like my job - I'm proud of what I do. I'm at the point in my life where I'm helping to find a cure for cancer instead of just seeing cancer specimens - and that's way cool. It's a great and honorable profession, and someone has to do it. I'm glad to be a part of it. Jackie O' Rene J Buesa Sent by: histonet-bounces@lists.utsouthwestern.edu 02/21/2006 10:21 AM To: pbrask@comcast.net, histonet@lists.utsouthwestern.edu cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: Re: [Histonet] Do you love your job? Hi Peggy: What others love or hate about a job has no individual relevance at all. You will have to experience that feeling by yourself. If you have dexterity, if you like to cook, if you like to knit, if you like to innovate and experiment, if you have a good smell sense, if you enjoy colours, if you look at what you manually have done and feel proud about it, if you understand the importance of doing things to help others, if you don't mind working more than required just to make sure that what you are doing is finished correctly, if you like at least some of the aforesaid, you will like histotechnology, if not, you are in a wrong switch from your previous activities. At least, those are some of the things I love; there are more, and what I don't like mostly are things related with human nature, and those you will find and hate in any profession. Hope this will give you an idea! ren? J. Peggy Brask wrote: I am a second semester student at Argosy University in the HT program. I was wondering what the pros and cons are in the Histonet world. I have been in the service industry in one form or another for the last 30 years. This is a total diversion from my past life and I would like to know what you love about your job and what you hate about your job. If anyone would like to give me some input, I would greatly appreciate it. Thanks, Peggy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Relax. Yahoo! Mail virus scanning helps detect nasty viruses! _______________________________________________ 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 End of Histonet Digest, Vol 27, Issue 31 **************************************** From jtrynak <@t> hotmail.com Wed Feb 22 06:39:34 2006 From: jtrynak <@t> hotmail.com (John Rynak) Date: Wed Feb 22 06:39:46 2006 Subject: [Histonet] Hiring -Cytopathology/Cytogenetics CLIA LAB - Boston Area Message-ID: Yes a bit off the normal mark - Just posting for a friend - Clinical Laboratory - Molecular Cytogenetics Living Microsystems is a startup company in Boston, MA, seeking exceptional talent to introduce its non-invasive technology that will provide women with accurate, and comprehensive prenatal clinical knowledge in early pregnancy. We are currently recruiting a team to establish our Cytogenetics Laboratory. We are looking for a Director, a Senior Scientist / Manager, and Cytogenetic Specialists. This team will provide fetal diagnostic information by performing molecular cytogenetic analysis of maternal blood samples. The team will also be involved with development of next generation products and analytic methods. Staff members will have experience in the following techniques: cell separation, FISH, PCR, DNA micro-arrays, computerized microscopy, cytogenetic analysis & scoring. Board certified candidates preferred. Interested candidates should send a resume and cover letter to: Human Resources Living Microsystems, Inc. 8 St. Marys Street Boston, MA 02215 Fax: (617) 353-1912 Email: hr@livingmicrosystems.com From dennijc <@t> vetmed.auburn.edu Wed Feb 22 07:44:42 2006 From: dennijc <@t> vetmed.auburn.edu (John C. Dennis) Date: Wed Feb 22 07:44:52 2006 Subject: [Histonet] PGP 9.5 In-Reply-To: <001b01c63747$7ede4760$ede49440@HPPav2> References: <001b01c63747$7ede4760$ede49440@HPPav2> Message-ID: Dear Peggy I've used Chemicon's rabbit polyclonal to good effect. It was used in the sniff trade for a while to mark vomeronasal sensory neurons, and processes, in particular. In double labeled preps, I found that, in at least several primate species and in dogs, not all sensory neurons in either the main olfactory or the vomeronasal epithelia express PGP9.5 in any given tissue section. Or at least at detectable levels. You'll see some or even most but not all neurons in a section. PGP 9.5 is a house keeping protein and is presumably up and down regulated depending on metabolic requirements. John Carroll Dennis Anatomy, Physiology, and Pharmacology 109 Greene Hall Auburn University, AL 36849 On Tue, 21 Feb 2006, Lee & Peggy Wenk wrote: > Anyone have experience with PGP 9.5 (Protein Gene Product)? It demonstrates > nerve fibers. One of our pathologists wants to use it on skin biopsies for > diagnosis of neuropathies. > > We would prefer peroxidase for paraffin sections, rather than FITC or FS. > Just wondering if people are having better luck with monoclonals or > polyclonals, and if there is a difference in animal species for the primary > (we'll be doing this on human skin). > > Also - all the papers we find mention thick sections and using a confocal > microscope. Anyone doing PGP on 5 um sections and/or with a regular light > microscope? > > One of my students is doing this as her research project. She's done a lot > of reading on it, and most papers don't discuss the actual procedure or the > type of antibody, etc. They are on the research aspect, or the diagnostic > aspect, not on what histotechs need to do the procedures. Those articles > that do mention techniques have very different methods (of course), and are > missing important information needed by histotechs. > > We would appreciate a couple of little hints of what works for "real life > histotechs", to get us started in the right direction for use in a routine > clinical immunohistochemistry lab. > > Feel free to contact me directly. Vendors included. I'll summarize and relay > the info back to Histonet. > > Thanks in advance. > > Peggy A. Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From JMahoney <@t> alegent.org Wed Feb 22 07:53:12 2006 From: JMahoney <@t> alegent.org (Janice Mahoney) Date: Wed Feb 22 07:54:10 2006 Subject: [Histonet] Opening IHC Message-ID: We have an immediate opening for a Histo Tech for the position of Immmunohistochemistry Resource Specialist. The right candidate will perform and oversee the IHC section of the our Histology laboratory. We perform approx 800 IHC slides per month and are growing this area. We are a LEAN laboratory with state of the art IHC instrumentation. Apply on-line at www.alegent.com We are an equal opportunity employer with a full benefit package. You may also call our Human Resources department at 402-593-3274. Janice Mahoney Histology/Cytology Coordinator Alegent Health Laboratory 4955 F Street Omaha, NE 68117 (402)717-2889 From rjbuesa <@t> yahoo.com Wed Feb 22 07:55:30 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Feb 22 07:55:40 2006 Subject: [Histonet] microwaves In-Reply-To: Message-ID: <20060222135530.71390.qmail@web61217.mail.yahoo.com> Jeanette: I used to work with a common household type microwave, but you will need to calibrate it. I am sending to your e-mail address a paper of mine dealing with this issue that I hope will be useful for your work. Ren? J. "Mitchell, Jeannette M." wrote: Hi Folks, What is everyone using when doing special stains for a microwave. Is there any OSHA regulations for microwaves??? Thanks for you help. Jeannette Mitchell Burlington, Vt Confidentiality Notice: This message, and any attachments, may contain information that is confidential, privileged, and/or protected from disclosure under state and federal laws that deal with the privacy and security of medical information. If you received this message in error or through inappropriate means, please reply to this message to notify the Sender that the message was received by you in error, and then permanently delete this message from all storage media, without forwarding or retaining a copy. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Autos. Looking for a sweet ride? Get pricing, reviews, & more on new and used cars. From rsrichmond <@t> aol.com Wed Feb 22 08:58:36 2006 From: rsrichmond <@t> aol.com (rsrichmond@aol.com) Date: Wed Feb 22 08:58:48 2006 Subject: [Histonet] Re: wright-giemsa stain on tissue In-Reply-To: <200602220727.82243fc58a91fb@rly-yj06.mx.aol.com> References: <200602220727.82243fc58a91fb@rly-yj06.mx.aol.com> Message-ID: <8C805D7979DA5D7-121C-9A0A@mblk-d18.sysops.aol.com> Dina (not otherwise identified): asks: >>Has anyone ever heard of staining wright-giemsa stain on paraffin embedded tissue (rat bone)? I've stained in the past blood smears and bone marrow smears. - Just wondering if anyone out there in histoland has ever done so. I checked out Histonet archives and did not find any results of paraffin processed tissue stained with Wright-Giemsa.<< Grumpy old pathologist time. Romanovsky type stains (Wright, Giemsa, what have you) are very useful with bone marrow sections, including decalcified material, but the necessary fixation is no longer practical. Tissue must be fixed with mercury and chromium (Zenker's or Helly's fixatives). The Wolbach Giemsa technique (actually developed for looking at rickettsiae - it's in the old AFIP manual) then prescribes staining in three or four changes of stain, one of them overnight, followed by differentiation in 10% colophonium rosin (abietic acid) in alcohol, using a microscope. I last saw such a stain in 1968, when I was a resident at Cornell Medical Center (can't remember what it's called this week) in New York City. Grump. Bob Richmond Gastonia NC From dellav <@t> musc.edu Wed Feb 22 09:22:51 2006 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Wed Feb 22 09:25:46 2006 Subject: [Histonet] (no subject) Message-ID: you will want to get your hands on the new NCCLS Standard for microwave use in laboratories, and also look at the new AP checklist from CAP, which can be downloaded from their website. these agencies have requirements that cover OSHA and FDA requirements for microwave use. Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> "Mitchell, Jeannette M." 02/22/06 06:33AM >>> Hi Folks, What is everyone using when doing special stains for a microwave. Is there any OSHA regulations for microwaves??? Thanks for you help. Jeannette Mitchell Burlington Confidentiality Notice: This message, and any attachments, may contain information that is confidential, privileged, and/or protected from disclosure under state and federal laws that deal with the privacy and security of medical information.. If you received this message in error or through inappropriate means, please reply to this message to notify the Sender that the message was received by you in error, and then permanently delete this message from all storage media, without forwarding or retaining a copy. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Wed Feb 22 09:42:48 2006 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Wed Feb 22 09:43:32 2006 Subject: [Histonet] Help for Transcription Message-ID: Good luck. We have three transciptionists for a Department of 16 pathologists. We do over 36,000 surgicals, almost 6,000 non-GYN cytologies, 8,000 Hematopathology accessions, and 2,000 outside consults. Many of the pathologists type their own reports or use "Dragon" voice recognition. Transcriptionists (like Histotechnologists) are an endangered species. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Weems, Joyce" 02/21/06 06:42PM >>> I need to justify additional transcriptionist hours. Would you please do me a favor and ask your transcription department for the number of minutes they have transcribed for Feb to date? - This can be found in the transcription system, I assume. We have Lanier and we have access to that info. I'd like to know # of cases Number of gross minutes dictated: Number of micro minutes dictated: Number of transcriptionists: Thanks for your help! Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From donna <@t> milestonemed.com Wed Feb 22 10:25:03 2006 From: donna <@t> milestonemed.com (Donna Willis) Date: Wed Feb 22 10:25:27 2006 Subject: [Histonet] microwaves In-Reply-To: Message-ID: <1140625514_6607@mail.serverlogistics.com> Jeannette, Yes there are regulations. Under Writer's Laboratory 923 states: "Use this appliance only for its intended use as described in the manual. Do not use corrosive chemicals or vapors in this appliance. This type of oven is specifically designed to heat, cook, or dry food, it is not designed for industrial or laboratory use." This is the statement that is sent with the warranty on household microwave. OHSA 29 CFR 1910.303(b)(2) states: "listed or labeled equipment shall be used or installed in accordance with any instrument included in the listing or labeling." This would include all equipment that is used in a laboratory setting that is not designed for laboratory use. If you have any other questions, let me know. Donna Willis, HT/HTL(ASCP) Milestone Medical North American Application Manager -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mitchell, Jeannette M. Sent: Wednesday, February 22, 2006 5:31 AM To: Histonet@pathology.swmed.edu Subject: [Histonet] microwaves Hi Folks, What is everyone using when doing special stains for a microwave. Is there any OSHA regulations for microwaves??? Thanks for you help. Jeannette Mitchell Burlington, Vt Confidentiality Notice: This message, and any attachments, may contain information that is confidential, privileged, and/or protected from disclosure under state and federal laws that deal with the privacy and security of medical information. If you received this message in error or through inappropriate means, please reply to this message to notify the Sender that the message was received by you in error, and then permanently delete this message from all storage media, without forwarding or retaining a copy. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Dorothy.L.Webb <@t> HealthPartners.Com Wed Feb 22 11:11:43 2006 From: Dorothy.L.Webb <@t> HealthPartners.Com (Webb, Dorothy L) Date: Wed Feb 22 11:12:44 2006 Subject: [Histonet] processing Message-ID: <0E394B648E5284478A6CCB78E5AFDA270171FC9D@hpes1.HealthPartners.int> I am submitting this for a colleague at another institution who has a concern over GI biopsies. They have 2 processors they use, one VIP and one Renaissance, both programmed with the same times and the same reagents at each station. They use recycled formalin. Their pathologists are complaining that the Gi biopsies that come off of the Renaissance processor are not of good quality, chattered, drier, etc. and the problems are not seen on the slides of the biopsies from the other (VIP) processor. They had a vendor out to check out the processor in question, inasmuch as they could be getting carry over of reagents, etc. and all checked out ok. Any suggestions as to the cause of their problem? Thanks ahead of time from a lead tech who is pulling out her hair!!!! I told her to "hold on", as my fellow histonetters would come up with something!! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From rjbuesa <@t> yahoo.com Wed Feb 22 11:20:07 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Feb 22 11:20:15 2006 Subject: [Histonet] processing In-Reply-To: <0E394B648E5284478A6CCB78E5AFDA270171FC9D@hpes1.HealthPartners.int> Message-ID: <20060222172007.5657.qmail@web61219.mail.yahoo.com> That is why I only have used and recommend VIP tissue processors! Just one suggestion: why they cannot process all the Bx in the VIP? And another: how about tissue samples other the GI Bx? If the other samples are processed OK with both tissue processors and it is a problem of reception time for the GI Bx, maybe they can run them with 2 short cycles/daily in the VIP. Just a thought! Ren? J. "Webb, Dorothy L" wrote: I am submitting this for a colleague at another institution who has a concern over GI biopsies. They have 2 processors they use, one VIP and one Renaissance, both programmed with the same times and the same reagents at each station. They use recycled formalin. Their pathologists are complaining that the Gi biopsies that come off of the Renaissance processor are not of good quality, chattered, drier, etc. and the problems are not seen on the slides of the biopsies from the other (VIP) processor. They had a vendor out to check out the processor in question, inasmuch as they could be getting carry over of reagents, etc. and all checked out ok. Any suggestions as to the cause of their problem? Thanks ahead of time from a lead tech who is pulling out her hair!!!! I told her to "hold on", as my fellow histonetters would come up with something!! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Autos. Looking for a sweet ride? Get pricing, reviews, & more on new and used cars. From jkiernan <@t> uwo.ca Wed Feb 22 11:55:14 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Wed Feb 22 11:55:25 2006 Subject: [Histonet] wright-giemsa stain on tissue References: <1140541481.43fb4829095a0@webmail.tulane.edu> Message-ID: <43FCA582.FA66FF6C@uwo.ca> Yes. Such methods have been available for many years. The older ones are somewhat complicated to do, and require fixation of the tissue in Helly's or other mercury-containing mixtures. A simplified method introduced by R.D.Lillie in 1943 (Public Health Reports 58: 449-452) and refined in 1944 (J. Tech. Methods Bull. Intl Assoc. Med. Museums 24:43). If you use an azure-eosin (blood) stain on sections of fixed tissue it is necessary to adjust the pH. Methanol-fixed blood smears are typically stained at pH 6.8, but for formaldehyde-fixed paraffin sections the pH must be lower, in the range 4 to 5. Detailed instructions are given in Lillie & Fullmer (1976), p.195-197. Lillie, who was a pathologist, preferred this type of method to H&E for routine use in the Technicon staining equipment of his day. I have used it (manually) and agree with Lillie that it's a much more informative staining technique than H&E. The pH is critical, however, and it's necessary to do tests in the range 3.5 to 5.5 to find the optimum for the material you are staining. Lillie & Fullmer provide some guidance on the relation of pH to type of fixation. pH 4 is often optimal for tissue that has been properly fixed (overnight or longer) in formaldehyde. For a more recent account, see Wittekind, DH et al. (1991) The standard Romanowsky-Giemsa stain in histology. Biotechnic & Histochemistry 66:282-295. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ dgaupp@tulane.edu asked: > > Histonet: > > Has anyone ever heard of staining wright-giemsa stain on paraffin embedded > tissue(rat bone)? I've stained in the past blood smears and bone marrow > smears. > Just wondering if anyone out there in histoland has ever done so. I checked out > histonet archives and did not find any results of paraffin processed tissue > stained with wright-giemsa. > > Thanks, > > Dina > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ploykasek <@t> phenopath.com Wed Feb 22 12:03:53 2006 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Wed Feb 22 12:04:17 2006 Subject: [Histonet] Do you love your job? In-Reply-To: Message-ID: Peggy - Kudos to you for venturing out into a different field that is new to you. I think histology is extremely interesting and challenging. As with anything, I think you get out of it what you put into it. I have been lucky to work with knowledgeable, generous, and kind pathologists. I went through a good program that taught me the basics, and have been taught much more by the pathologists I've worked with over the years. I have a very curious nature, and am always finding out something new - some new fact, a new way to do something, a new antibody to work up, etc... Salaries have greatly increased since I first entered the field. I think it is appropriate for the responsibilities that we have for the specimens. As for the cons, I'm trying to think of some. Sometimes the work hours are a bit weird. Some doctors are a bit challenging to work with - I try to adjust how I approach working with them & usually it works out. The regulations sometimes make me crazy, but I do my best to learn them & comply. Along with the regulations comes the paperwork. I want to know what happened to the computer age of a 'paperless' system! At any rate, good luck with your new career. I hope that you enjoy it. Patti Loykasek BS, HTL, QIHC PhenoPath Laboratories Seattle, WA > I am a second semester student at Argosy University in the HT program. I > was wondering what the pros and cons are in the Histonet world. I have been > in the service industry in one form or another for the last 30 years. This > is a total diversion from my past life and I would like to know what you > love about your job and what you hate about your job. If anyone would like > to give me some input, I would greatly appreciate it. > > Thanks, Peggy > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------- This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. From Eric.C.Kellar <@t> questdiagnostics.com Wed Feb 22 12:05:11 2006 From: Eric.C.Kellar <@t> questdiagnostics.com (Kellar, Eric C) Date: Wed Feb 22 12:05:35 2006 Subject: [Histonet] processing Message-ID: <6843061CE6B98E4B96590D4F299618F801583BAD@qdcws0117.us.qdx.com> Dorothy, I infiltrate my GI biopsies with a low melting point paraffin (Paraplast Xtra) at 58*C on my VIP processors. GI biopsies easily become friable at paraffin temperatures of 62*C and over. Ask her to check the temps of her paraffins between the two processors. Also, GI biopsies seem to be more challanging to cut after fresh dehydrating alcohol solutions are added to the processors. They respond well to graduated alcohols from 70%, 80%, 95%, 100% to the clearant. I do not use the Ventana processor, but I do know it does require some fine tuning to program the dehydrating alcohols to match those on the VIP. Eric C. Kellar Histology/Immunohistochemistry Quest Diagnostics Inc. Miami -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Webb, Dorothy L Sent: Wednesday, February 22, 2006 12:12 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] processing I am submitting this for a colleague at another institution who has a concern over GI biopsies. They have 2 processors they use, one VIP and one Renaissance, both programmed with the same times and the same reagents at each station. They use recycled formalin. Their pathologists are complaining that the Gi biopsies that come off of the Renaissance processor are not of good quality, chattered, drier, etc. and the problems are not seen on the slides of the biopsies from the other (VIP) processor. They had a vendor out to check out the processor in question, inasmuch as they could be getting carry over of reagents, etc. and all checked out ok. Any suggestions as to the cause of their problem? Thanks ahead of time from a lead tech who is pulling out her hair!!!! I told her to "hold on", as my fellow histonetters would come up with something!! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ============================================================================== The contents of this message, together with any attachments, are intended only for the use of the person(s) to which they are addressed and may contain confidential and/or privileged information. Further, any medical information herein is confidential and protected by law. It is unlawful for unauthorized persons to use, review, copy, disclose, or disseminate confidential medical information. If you are not the intended recipient, immediately advise the sender and delete this message and any attachments. Any distribution, or copying of this message, or any attachment, is prohibited. ============================================================================== From nadams <@t> CapeCodHealth.org Wed Feb 22 12:09:31 2006 From: nadams <@t> CapeCodHealth.org (Adams, Nancy) Date: Wed Feb 22 12:09:40 2006 Subject: [Histonet] non mercury calibrating thermometer Message-ID: <17A1862099540D458C8FE9380C2BC461C09338@fh2xmail.fhdomain1.capecodhealth.org> Looking for place to purchase non mercury calibrating thermometer. Anyone? Thanks Nancy Rutledge ************************************************************ This email and any files transmitted with it are confidential. And intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please contact the system administrator for Cape Cod Healthcare. Administrator@CapeCodHealth.org ************************************************************ From TJasper <@t> smdc.org Wed Feb 22 12:14:02 2006 From: TJasper <@t> smdc.org (Jasper, Thomas G.) Date: Wed Feb 22 12:13:49 2006 Subject: [Histonet] processing Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA07D39159@harrier> Hey Dorothy, Since she has 2 machines why not have her program one for small biopsies etc. I don't know what her normal runs are set at, but we run 2 processors, one for small biopsies only. The small run is set up to be shorter and has no heat (European colleagues, for the most part question this heat practice, but I digress). I would think she would eliminate her problem with the shorter, gentler run. You have to be careful during the day when the tissue is grossed in. The PAs a pathologists need to know about the separation. I certainly don't think getting the pathologists on board for this is a hard sell. Good luck! tj Thomas Jasper HT(ASCP)BAS Anatomic Pathology Supervisor SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Webb, Dorothy L Sent: Wednesday, February 22, 2006 11:12 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] processing I am submitting this for a colleague at another institution who has a concern over GI biopsies. They have 2 processors they use, one VIP and one Renaissance, both programmed with the same times and the same reagents at each station. They use recycled formalin. Their pathologists are complaining that the Gi biopsies that come off of the Renaissance processor are not of good quality, chattered, drier, etc. and the problems are not seen on the slides of the biopsies from the other (VIP) processor. They had a vendor out to check out the processor in question, inasmuch as they could be getting carry over of reagents, etc. and all checked out ok. Any suggestions as to the cause of their problem? Thanks ahead of time from a lead tech who is pulling out her hair!!!! I told her to "hold on", as my fellow histonetters would come up with something!! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. As required by federal and state laws, you need to hold this information as privileged and confidential. If you have received this communication in error, please notify the sender and destroy all copies of this communication and any attachments. From perryg <@t> uthscsa.edu Wed Feb 22 12:16:53 2006 From: perryg <@t> uthscsa.edu (Perry, Griffin M) Date: Wed Feb 22 12:16:58 2006 Subject: [Histonet] Leica CM1800 Disposable Blade Holder Message-ID: <39C2F40A8A1EA646AF1A6F187698656402BB4505@NYALA.win.uthscsa.edu> Dear Histonet Users, I have a Leica CM1800. We're looking to switch over and use disposable blades. I know we need a blade holder...is there like a universal disposable blade holder for Leicas or Cryostats in general? What about disposable blades? I've been hearing good things about Accu-Edge Lo-Pro Blades. I'd appreciate any feedback. Thanks, G From ploykasek <@t> phenopath.com Wed Feb 22 11:41:49 2006 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Wed Feb 22 12:32:46 2006 Subject: [Histonet] Galectin antibody Message-ID: Is anyone using a galectin-3 antibody clone 9C4 from Novocastra in paraffin sections? We have been working this antibody up with ok results but would like to try to clean up the signal a bit. We are using a heat antigen retrieval with EDTA. Just wondering what others are using & their experience with this antibody. Thank you. Patti Loykasek BS, HTL, QIHC PhenoPath Laboratories Seattle, WA ------------------------------------------------------------------------- This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. From TJJ <@t> Stowers-Institute.org Wed Feb 22 13:07:04 2006 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Wed Feb 22 13:07:36 2006 Subject: [Histonet] Re: Wright-Giemsa stain on tissue Message-ID: Dina, In my days in clinical labs, we did a May-Grunwald Giemsa stain on our bone marrow biopsies and particle preps. I'm with Dr. Richmond in that there the staining results from the Zenker's fixed material was second to none. However, we were able to do a reasonably good job of it, provided the pathologists never had to pull archival slides to compare. That being said, I am happy to never go back to using it, what a pain! You might try using the M-G Giemsa technique and see how it works for you. Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From TJJ <@t> Stowers-Institute.org Wed Feb 22 13:20:26 2006 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Wed Feb 22 13:20:58 2006 Subject: [Histonet] Re: Sucrose cryosectioning Message-ID: Weird, but I have never experienced difficulty with sectioning sucrose cryopreserved material. I've heard Gayle Callis mention how awful it is, and now others say they have problems with it. (I'm almost afraid to speak up that I may jinx myself!) I agree with Andi, it's a good idea to allow the sample to sit in OCT for a bit before freezing, but on some of our chick and mouse embryos, we have embedded in OCT and then quickly frozen with good results. We have sectioned mouse testis, femur, eyes, and pancreas all treated this way and never had an ooey gooey mess, even at -20 to -25 C. For the larger adult samples, I think you probably do need to have the cryostat colder than you would for unfixed frozen samples. We formalin fix 2 hours for small samples, to overnight for large samples, then rinse in PBS, place into 15% sucrose until they sink, then place into 30% sucrose until they sink. Remove as much of the sucrose as you can (briefly blot adult samples) before placing into OCT, and allow to sit up to an hour before freezing (cover, so the OCT doesn't harden!) Pancreas has turned out to be the worst in terms of morphology for us. Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From Rcartun <@t> harthosp.org Wed Feb 22 13:43:03 2006 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Wed Feb 22 13:43:52 2006 Subject: [Histonet] Oct-3/4 Message-ID: This is what we do: Antibody name: Oct 3/4 (C-20) - Source: Santa Cruz Biotechnology (Cat #SC 8629) - Cat #: SC-8629 - Made in: Goat - Pretreatment: HIER (vegetable steamer) for 20 min. with citrate buffer pH6 (DAKO) - Antibody dilution: 1:100 - Detection System: Rabbit anti-Goat-HRP; diluted 1:100 (DAKO) - Chromogen: DAB+ (Dako) Staining is performed on a DAKO Autostainer. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Elaine Dooley" 02/20/06 02:51PM >>> Dear Histonetters, I have been having some trouble trying to get this antibody to stain seminomas. It is Oct-3/4 a goat polyclonal antibody from Santa Cruz Biotechnology. We also purchased their mouse anti-goat biotin conjugated secondary. Any quick hints on making this work. I have tried it on the Ventana Benchmark (with changing the secondary antibody). I have tried overnight incubations and staining entirely by hand. I have tried Ventana's CC1 and also tried Cell Marque's trilogy for epitope retrial but have not had any of the desired nuclear staining the antibody is supposed to show. Any helpful hints would be greatly appreciated. Elaine Dooley HTL Shands Teaching Hospital Gainesville FL 352-265-0111 ext 7-2117 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SAllen <@t> exchange.hsc.mb.ca Wed Feb 22 14:20:13 2006 From: SAllen <@t> exchange.hsc.mb.ca (Sharon Allen) Date: Wed Feb 22 14:21:45 2006 Subject: [Histonet] disposal of xylol waste from CJD cases Message-ID: <36FE435D6D2F1D489174B22362A961B66B8319@hscxntmx0006.hsc.mb.ca> I am having a problem disposing or our xylol waste from Prion testing for CJD. Our waste disposal company has just informed us thay they will take all other waste but won't touch the xylol (it has all been treated with javex). Apparently they have been storing it all these years & now are refusing to accept anymore. I would like to know how other labs testing for CJD are disposing of their xylol. Does anyone use a xylol substitute? Any ideas would be very helpful. Right now I have it packaged & sitting on the floor in the lab which goes against all the safety rules. Sharon sallen@hsc.mb.ca This e-mail and/or any documents in this transmission is intended for the address(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. From maria <@t> ski.org Wed Feb 22 14:53:09 2006 From: maria <@t> ski.org (Maria Mejia) Date: Wed Feb 22 14:53:20 2006 Subject: [Histonet] Re: Sucrose cryosectioning References: Message-ID: <43FCCF35.8030209@ski.org> I too have not experienced any difficulty with sectioning sucrose cryo material. For the past 16 years, I have sectioned feline & primate cortical tissue - some small and others very large on the sliding microtome. Both animals are perfused. The cortical tissue of interest is removed & tissue is then further fixed overnight @ 4C. Next day, tissue is rinsed in PBS, placed in 30% sucrose/0.1M PB at 4C until the tissue sinks to bottom of container. I usually leave in this solution for another day & equally important I change the solution once a day! After which the tissue is briefly blotted and placed into OCT before freezing and sectioned. Although, I have not left my sample sit in OCT for up to an hour - I do leave the sample in 4 degree OCT for at least 15-20 minutes. Hope this helps. Maria Bartola Mejia Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 Email: maria@ski.org or mbmphoto@gmail.com Phone: (415)-621-8242 Johnson, Teri wrote: >Weird, but I have never experienced difficulty with sectioning sucrose >cryopreserved material. I've heard Gayle Callis mention how awful it >is, and now others say they have problems with it. (I'm almost afraid to >speak up that I may jinx myself!) > > I agree with Andi, it's a good idea to allow the sample to sit in OCT >for a bit before freezing, but on some of our chick and mouse embryos, >we have embedded in OCT and then quickly frozen with good results. > >We have sectioned mouse testis, femur, eyes, and pancreas all treated >this way and never had an ooey gooey mess, even at -20 to -25 C. For >the larger adult samples, I think you probably do need to have the >cryostat colder than you would for unfixed frozen samples. > >We formalin fix 2 hours for small samples, to overnight for large >samples, then rinse in PBS, place into 15% sucrose until they sink, then >place into 30% sucrose until they sink. Remove as much of the sucrose as >you can (briefly blot adult samples) before placing into OCT, and allow >to sit up to an hour before freezing (cover, so the OCT doesn't harden!) > >Pancreas has turned out to be the worst in terms of morphology for us. > >Teri Johnson, HT(ASCP)QIHC >Managing Director Histology Facility >Stowers Institute for Medical Research >1000 E. 50th St. >Kansas City, MO 64133 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From Rcartun <@t> harthosp.org Wed Feb 22 15:13:08 2006 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Wed Feb 22 15:14:54 2006 Subject: [Histonet] A call to action for US AP/Histology professionals Message-ID: One way or another, we must cut spending. Health care costs are out of control (16% of our economic output is consumed by health care). Our country cannot continue down this path. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> Cheryl 02/21/06 10:00AM >>> Hello Folks-- There is another issue we should consider a call to paper the Senate with letters stating our opinions....The AFIP faces closure due to realignment by base closures (BRAC)--this includes the pathology center AND the school!! We've lost so many good schools, and the AFIP is among the best. There is a bill to relocate core services of the AFIP to another center and the link below provides an easy way to write you your senator in support of this bill: http://capwiz.com/ascpath/issues/alert/?alertid=8171231&type=CO Please take a minute to show your support for our science on MUE billing (Peggy's email of earlier today),AFIP closure, and two additional issues threatening continued viability of lab science in this country. The form is easy to fill out, and you can forward the link to several friends once you submit your letter (it will give you another pop-up for sharing the letter once you click to submit yours) to increase the impact of our unified voice. This link will lead you to both letters and you can send it by email or snail mail: http://capwiz.com/ascpath/home/ Thank you for your continued interest and support tp maintain visibility and viability of our craft. Together we DO make a difference. Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one Tech at a time. 281.883.7704 c 281.852.9457 o admin@fullstaff.org Sign up for the FREE monthly newsletter AP News--updates, tricks of the trade and current issues for Anatomic Pathology Clinical Labs. Send a 'subscribe' request to APNews@fullstaff.org. Please include your name and specialty in the body of the email. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Wed Feb 22 15:27:50 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Wed Feb 22 15:27:59 2006 Subject: [Histonet] A call to action for US AP/Histologyprofessionals Message-ID: <83AACDB0810528418AA106F9AE9B7F7E013058AB@sjhaexc02.sjha.org> This is true, but it should be a fair thing. Why should a dermatologist collect for 10 reimbursements and the pathologist only two of those ten? j -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard Cartun Sent: Wednesday, February 22, 2006 4:13 PM To: Histonet; Cheryl Subject: Re: [Histonet] A call to action for US AP/Histologyprofessionals One way or another, we must cut spending. Health care costs are out of control (16% of our economic output is consumed by health care). Our country cannot continue down this path. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> Cheryl 02/21/06 10:00AM >>> Hello Folks-- There is another issue we should consider a call to paper the Senate with letters stating our opinions....The AFIP faces closure due to realignment by base closures (BRAC)--this includes the pathology center AND the school!! We've lost so many good schools, and the AFIP is among the best. There is a bill to relocate core services of the AFIP to another center and the link below provides an easy way to write you your senator in support of this bill: http://capwiz.com/ascpath/issues/alert/?alertid=8171231&type=CO Please take a minute to show your support for our science on MUE billing (Peggy's email of earlier today),AFIP closure, and two additional issues threatening continued viability of lab science in this country. The form is easy to fill out, and you can forward the link to several friends once you submit your letter (it will give you another pop-up for sharing the letter once you click to submit yours) to increase the impact of our unified voice. This link will lead you to both letters and you can send it by email or snail mail: http://capwiz.com/ascpath/home/ Thank you for your continued interest and support tp maintain visibility and viability of our craft. Together we DO make a difference. Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one Tech at a time. 281.883.7704 c 281.852.9457 o admin@fullstaff.org Sign up for the FREE monthly newsletter AP News--updates, tricks of the trade and current issues for Anatomic Pathology Clinical Labs. Send a 'subscribe' request to APNews@fullstaff.org. Please include your name and specialty in the body of the email. _______________________________________________ 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 Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From lkbauer <@t> unmc.edu Wed Feb 22 15:57:30 2006 From: lkbauer <@t> unmc.edu (Linda K Bauer) Date: Wed Feb 22 15:57:40 2006 Subject: [Histonet] removing excess x-gal stain Message-ID: I would like to remove excess nonspecific x-gal stain from whole mount murine embryos. Does anyone have a suggestion or technique for this? Linda(Lin)Bauer Department of Genetics, Cell Biology, and Anatomy 985455 Nebraska Medical Center Omaha, NE 68198-5455 Phone: (402) 559-2863 Fax: (402) 559-4001 Email: lkbauer@unmc.edu From jcolclefa <@t> aol.com Wed Feb 22 16:04:12 2006 From: jcolclefa <@t> aol.com (John PJ Coleman) Date: Wed Feb 22 16:04:32 2006 Subject: [Histonet] Microwave processing Message-ID: I am the Senior tech of a large hospital corporation. My administration has just won funding for 4 Sakura Microwave rapid processing units. We run FISH her 2 on formalin fixed paraffin embedded tissue as per FDA protocol. As I tech, I am not in favor of tossing routine processing wholesale in favor of a completely new technology without thorough testing and parallel processing. Also, we are a regional reference lab for IHC and have a panel of 115 antibodies, all optimized for formalin fixed paraffin embedded tissue. We run an average of 350 IHC slides a day, max 580 per day and would have to re-optimize these to use in the new formalin free world while keeping our FFPE procedures in parallel for our reference lab work. Much like running 2 labs. If anyone has any insight, or if anyone currently uses these instruments for routine and/or IHC, feel free to call or email, and I'll check the postings on this string. We are also taking invitations to come out and see these things in use real time. John PJ Coleman-757 335-2159 http://members.cox.net/captainmadman/ http://journals.aol.com/jcolclefa/Waytoomuchboringcraptosayusually/ From LuckG <@t> empirehealth.org Wed Feb 22 16:28:17 2006 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Wed Feb 22 16:28:29 2006 Subject: [Histonet] Microwave processing Message-ID: John, Your concerns are very valid particularly in your role as a reference lab. A couple of questions. 1st, which part of "FFPE" does the Sakura instrument not fulfill (e.g. are the tissues not exposed to NBF during the Sakura processing cycle). 2nd, When you perform IHC in the role as a reference lab how are you currently ascertaining how the paraffin blocks you receive for IHC have been processed? In a somewhat analogous situation for years we maintained two IHC protocols because we did histology for two hospitals. One hospital used NBF as its designated routine fixative and the other used "Prefer". Was somewhat of a pain but was manageable, however we only average about 50-60 IHC's/day. I could share some of what we learned developing and maintaining two standing IHC protocols for each antibody if you think our situation has any potential learning value for you. Good luck, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org www.deaconessmedicalcenter.org -----Original Message----- From: John PJ Coleman [mailto:jcolclefa@aol.com] Sent: Wednesday, February 22, 2006 2:04 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave processing I am the Senior tech of a large hospital corporation. My administration has just won funding for 4 Sakura Microwave rapid processing units. We run FISH her 2 on formalin fixed paraffin embedded tissue as per FDA protocol. As I tech, I am not in favor of tossing routine processing wholesale in favor of a completely new technology without thorough testing and parallel processing. Also, we are a regional reference lab for IHC and have a panel of 115 antibodies, all optimized for formalin fixed paraffin embedded tissue. We run an average of 350 IHC slides a day, max 580 per day and would have to re-optimize these to use in the new formalin free world while keeping our FFPE procedures in parallel for our reference lab work. Much like running 2 labs. If anyone has any insight, or if anyone currently uses these instruments for routine and/or IHC, feel free to call or email, and I'll check the postings on this string. We are also taking invitations to come out and see these things in use real time. John PJ Coleman-757 335-2159 http://members.cox.net/captainmadman/ http://journals.aol.com/jcolclefa/Waytoomuchboringcraptosayusually/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From igor.nasonkin <@t> jhmi.edu Wed Feb 22 16:40:22 2006 From: igor.nasonkin <@t> jhmi.edu (IGOR NASONKIN) Date: Wed Feb 22 16:40:31 2006 Subject: [Histonet] Re: Sucrose cryosectioning Message-ID: Maria, How do you recommend to freeze tissue after 30% sucrose and OCT imersion? Snap-freezing on dry ice or in methylbutane/fry ice bath? I've been doing mostly paraffin sectioning until now; as i see, people have different and strong opinions on how to freeze tissue post-sucrose. Most agree that methylbutane/dry ice is a good method to snap-freeze tissue; however i heard another, strong opinion, that this introduces some structural damage to tissue, and freezing in just dry ice is better. However, nobody around here immerses in OCT after sucrose, OCT is added later to hold the specimen. So, -how you freeze after sucrose-OCT. Thanks. Hope some comments will follow re. methylbutane as well. Igor ----- Original Message ----- From: Maria Mejia Date: Wednesday, February 22, 2006 3:53 pm Subject: Re: [Histonet] Re: Sucrose cryosectioning > I too have not experienced any difficulty with sectioning sucrose > cryo material. For the past 16 years, I have sect ioned feline & > primate > cortical > tissue - some small and others very large on the sliding microtome. > > Both animals are perfused. The cortical tissue of interest is > removed & > tissue > is then further fixed overnight @ 4C. Next day, tissue is rinsed in > PBS, > placed > in 30% sucrose/0.1M PB at 4C until the tissue sinks to bottom of > container. I > usually leave in this solution for another day & equally important > I > change the > solution once a day! > > After which the tissue is briefly blotted and placed into OCT > before > freezing and > sectioned. Although, I have not left my sample sit in OCT for up to > an > hour - I do > leave the sample in 4 degree OCT for at least 15-20 minutes. > > Hope this helps. > > Maria Bartola Mejia > Smith-Kettlewell Eye Research Institute > San Francisco, CA 94115 > Email: maria@ski.org or mbmphoto@gmail.com > Phone: (415)-621-8242 > > > > > > Johnson, Teri wrote: > > >Weird, but I have never experience d difficulty with sectioning > sucrose>cryopreserved material. I've heard Gayle Callis mention > how awful it > >is, and now others say they have problems with it. (I'm almost > afraid to > >speak up that I may jinx myself!) > > > > I agree with Andi, it's a good idea to allow the sample to sit in > OCT>for a bit before freezing, but on some of our chick and mouse > embryos,>we have embedded in OCT and then quickly frozen with good > results.> > >We have sectioned mouse testis, femur, eyes, and pancreas all treated > >this way and never had an ooey gooey mess, even at -20 to -25 C. For > >the larger adult samples, I think you probably do need to have the > >cryostat colder than you would for unfixed frozen samples. > > > >We formalin fix 2 hours for small samples, to overnight for large > >samples, then rinse in PBS, place into 15% sucrose until they > sink, then > >place into 30% sucrose until they sink. Remove as much of the > sucrose as > >you can (briefly blot a dult samples) before placing into OCT, and > allow>to sit up to an hour before freezing (cover, so the OCT > doesn't harden!) > > > >Pancreas has turned out to be the worst in terms of morphology for > us.> > >Teri Johnson, HT(ASCP)QIHC > >Managing Director Histology Facility > >Stowers Institute for Medical Research > >1000 E. 50th St. > >Kansas City, MO 64133 > > > >_______________________________________________ > >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 > From jdmd77 <@t> hotmail.com Wed Feb 22 17:00:45 2006 From: jdmd77 <@t> hotmail.com (Julia Dahl) Date: Wed Feb 22 17:01:00 2006 Subject: [Histonet] A call to action for US AP/Histologyprofessionals In-Reply-To: <83AACDB0810528418AA106F9AE9B7F7E013058AB@sjhaexc02.sjha.org> Message-ID: Thank you Joyce for your understanding of the issues at hand with the discrepant handling of MUEs for the people doing the procedure and those of us involved in making the diagnosis on the tissue procured from the procedure. If the MUE for a biopsy is TEN - then the MUE for the pathology services on biopsies should be TEN, not two. Yes, healthcare costs are rising and do need to be curtailed, blanket reduction in pathology services reimbursement out of proportion to other specialties - claiming that anything beyond two biopsies on a single patient on a single day are "unbelievable edits" so far outside the standard of care as to be considered equivalent to a typographical error is heinous. Every patient with chronic diarrhea, iron deficiency anemia, dysphagia, Barrett's esophagus, inflammatory bowel disease NEEDS to have more than two sites biopsied FOR the standard of care; every man with an elevated PSA, every woman with multiple breast lumps needs to have more than two sites biopsies FOR the standard of care to be maintained. Again, Joyce, thank you. The MUEs for pathology are not the answer to the current ails of the American Medical system. Julia Dahl, M.D. Pathologist >This is true, but it should be a fair thing. Why should a dermatologist >collect for 10 reimbursements and the pathologist only two of those ten? >j > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard >Cartun >Sent: Wednesday, February 22, 2006 4:13 PM >To: Histonet; Cheryl >Subject: Re: [Histonet] A call to action for US >AP/Histologyprofessionals > >One way or another, we must cut spending. Health care costs are out of >control (16% of our economic output is consumed by health care). Our >country cannot continue down this path. > >Richard > >Richard W. Cartun, Ph.D. >Director, Immunopathology & Histology >Assistant Director, Anatomic Pathology >Hartford Hospital >80 Seymour Street >Hartford, CT 06102 >(860) 545-1596 >(860) 545-0174 Fax > > >>> Cheryl 02/21/06 10:00AM >>> >Hello Folks-- > > There is another issue we should consider a call to paper the Senate >with letters stating our opinions....The AFIP faces closure due to >realignment by base closures (BRAC)--this includes the pathology center >AND the school!! We've lost so many good schools, and the AFIP is among >the best. There is a bill to relocate core services of the AFIP to >another center and the link below provides an easy way to write you your >senator in support of this bill: > > http://capwiz.com/ascpath/issues/alert/?alertid=8171231&type=CO > > Please take a minute to show your support for our science on MUE >billing (Peggy's email of earlier today),AFIP closure, and two >additional issues threatening continued viability of lab science in this >country. The form is easy to fill out, and you can forward the link to >several friends once you submit your letter (it will give you another >pop-up for sharing the letter once you click to submit yours) to >increase the impact of our unified voice. This link will lead you to >both letters and you can send it by email or snail mail: > http://capwiz.com/ascpath/home/ > > Thank you for your continued interest and support tp maintain >visibility and viability of our craft. Together we DO make a >difference. > > > >Cheryl Kerry, HT(ASCP) >Full Staff Inc. >Staffing the AP Lab by helping one Tech at a time. >281.883.7704 c >281.852.9457 o >admin@fullstaff.org > >Sign up for the FREE monthly newsletter AP News--updates, tricks of the >trade and current issues for Anatomic Pathology Clinical Labs. Send a >'subscribe' request to APNews@fullstaff.org. Please include your name >and specialty in the body of the email. >_______________________________________________ >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 > > >Confidentiality Notice ** The information contained in this message may be >privileged and is confidential information intended for the use of the >addressee listed above. If you are neither the intended recipient nor the >employee or agent responsible for delivering this message to the intended >recipient, you are hereby notified that any disclosure, copying, >distribution or the taking of any action in reliance on the contents of >this information is strictly prohibited. If you have received this >communication in error, please notify us immediately by replying to the >message and deleting it from your computer. Thank you. Saint Joseph's >Health System, Inc. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From JWEEMS <@t> sjha.org Wed Feb 22 17:06:09 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Wed Feb 22 17:06:17 2006 Subject: [Histonet] A call to action for US AP/Histologyprofessionals Message-ID: <83AACDB0810528418AA106F9AE9B7F7E013058AF@sjhaexc02.sjha.org> Thanks for explaining it better. We really do need to work together on this and we need to make it happen before July 1 and not let it hang around like the cyto proficiency testing which took a year of frayed nerves and thousands of dollars before it got held. -----Original Message----- From: Julia Dahl [mailto:jdmd77@hotmail.com] Sent: Wednesday, February 22, 2006 6:01 PM To: Weems, Joyce; Rcartun@harthosp.org; histonet@lists.utsouthwestern.edu; tkngflght@yahoo.com Subject: RE: [Histonet] A call to action for US AP/Histologyprofessionals Thank you Joyce for your understanding of the issues at hand with the discrepant handling of MUEs for the people doing the procedure and those of us involved in making the diagnosis on the tissue procured from the procedure. If the MUE for a biopsy is TEN - then the MUE for the pathology services on biopsies should be TEN, not two. Yes, healthcare costs are rising and do need to be curtailed, blanket reduction in pathology services reimbursement out of proportion to other specialties - claiming that anything beyond two biopsies on a single patient on a single day are "unbelievable edits" so far outside the standard of care as to be considered equivalent to a typographical error is heinous. Every patient with chronic diarrhea, iron deficiency anemia, dysphagia, Barrett's esophagus, inflammatory bowel disease NEEDS to have more than two sites biopsied FOR the standard of care; every man with an elevated PSA, every woman with multiple breast lumps needs to have more than two sites biopsies FOR the standard of care to be maintained. Again, Joyce, thank you. The MUEs for pathology are not the answer to the current ails of the American Medical system. Julia Dahl, M.D. Pathologist >This is true, but it should be a fair thing. Why should a dermatologist >collect for 10 reimbursements and the pathologist only two of those ten? >j > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard >Cartun >Sent: Wednesday, February 22, 2006 4:13 PM >To: Histonet; Cheryl >Subject: Re: [Histonet] A call to action for US >AP/Histologyprofessionals > >One way or another, we must cut spending. Health care costs are out of >control (16% of our economic output is consumed by health care). Our >country cannot continue down this path. > >Richard > >Richard W. Cartun, Ph.D. >Director, Immunopathology & Histology >Assistant Director, Anatomic Pathology >Hartford Hospital >80 Seymour Street >Hartford, CT 06102 >(860) 545-1596 >(860) 545-0174 Fax > > >>> Cheryl 02/21/06 10:00AM >>> >Hello Folks-- > > There is another issue we should consider a call to paper the Senate >with letters stating our opinions....The AFIP faces closure due to >realignment by base closures (BRAC)--this includes the pathology center >AND the school!! We've lost so many good schools, and the AFIP is among >the best. There is a bill to relocate core services of the AFIP to >another center and the link below provides an easy way to write you your >senator in support of this bill: > > http://capwiz.com/ascpath/issues/alert/?alertid=8171231&type=CO > > Please take a minute to show your support for our science on MUE >billing (Peggy's email of earlier today),AFIP closure, and two >additional issues threatening continued viability of lab science in this >country. The form is easy to fill out, and you can forward the link to >several friends once you submit your letter (it will give you another >pop-up for sharing the letter once you click to submit yours) to >increase the impact of our unified voice. This link will lead you to >both letters and you can send it by email or snail mail: > http://capwiz.com/ascpath/home/ > > Thank you for your continued interest and support tp maintain >visibility and viability of our craft. Together we DO make a >difference. > > > >Cheryl Kerry, HT(ASCP) >Full Staff Inc. >Staffing the AP Lab by helping one Tech at a time. >281.883.7704 c >281.852.9457 o >admin@fullstaff.org > >Sign up for the FREE monthly newsletter AP News--updates, tricks of the >trade and current issues for Anatomic Pathology Clinical Labs. Send a >'subscribe' request to APNews@fullstaff.org. Please include your name >and specialty in the body of the email. >_______________________________________________ >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 > > >Confidentiality Notice ** The information contained in this message may be >privileged and is confidential information intended for the use of the >addressee listed above. If you are neither the intended recipient nor the >employee or agent responsible for delivering this message to the intended >recipient, you are hereby notified that any disclosure, copying, >distribution or the taking of any action in reliance on the contents of >this information is strictly prohibited. If you have received this >communication in error, please notify us immediately by replying to the >message and deleting it from your computer. Thank you. Saint Joseph's >Health System, Inc. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From rjbuesa <@t> yahoo.com Wed Feb 22 17:06:36 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Feb 22 17:06:47 2006 Subject: [Histonet] Microwave processing In-Reply-To: Message-ID: <20060222230636.28591.qmail@web61219.mail.yahoo.com> John: I know this instrument and the technology is safe. You will not need to do HIER for IHC since the tissue will be fixed in an alcohol based fixatives and I don't think that you will need to run parallel runs.Maybe you will have to dilute your antibodies a little more than for formalin + HIER. The turn around time is 120 cassettes per hour, and with 4 of these instruments (which I personally think is a "bit too much") what you will have to do is to rethink your whole staff scheduling. Try to get a book titled: Microwaves for the Art of Microscopy by L.P Kok and M.E.Boon, Coulomb Press Leyden 2003 (you can get it at http://www.coulomb.nl This instrument is ideal for a large volume reference lab like yours. Hope this will help you. Ren? J. John PJ Coleman wrote: I am the Senior tech of a large hospital corporation. My administration has just won funding for 4 Sakura Microwave rapid processing units. We run FISH her 2 on formalin fixed paraffin embedded tissue as per FDA protocol. As I tech, I am not in favor of tossing routine processing wholesale in favor of a completely new technology without thorough testing and parallel processing. Also, we are a regional reference lab for IHC and have a panel of 115 antibodies, all optimized for formalin fixed paraffin embedded tissue. We run an average of 350 IHC slides a day, max 580 per day and would have to re-optimize these to use in the new formalin free world while keeping our FFPE procedures in parallel for our reference lab work. Much like running 2 labs. If anyone has any insight, or if anyone currently uses these instruments for routine and/or IHC, feel free to call or email, and I'll check the postings on this string. We are also taking invitations to come out and see these things in use real time. John PJ Coleman-757 335-2159 http://members.cox.net/captainmadman/ http://journals.aol.com/jcolclefa/Waytoomuchboringcraptosayusually/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Brings words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. From dholmes <@t> anatomy.umsmed.edu Wed Feb 22 17:08:28 2006 From: dholmes <@t> anatomy.umsmed.edu (Dianne Holmes) Date: Wed Feb 22 17:08:48 2006 Subject: [Histonet] Blue spots Message-ID: Help! I am seeing blue spots !!!? I have completed a BDA/DAB reaction on some rabbit brain tissue. When I used Cresyl Violet for a counterstain - I had small blue "ponds" randomly showing up on the tissue. I have used the same method for years and had not gotten this result? Basic info: tissue is paraformaldehyde fixed and cut as frozen sections @ 50 microns. Used same batch of CV that has been used in the past. The rest of the tissue looks perfectly fine. Any ideas as to what went wrong? From JWEEMS <@t> sjha.org Wed Feb 22 17:14:46 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Wed Feb 22 17:14:55 2006 Subject: [Histonet] Microwave processing Message-ID: <83AACDB0810528418AA106F9AE9B7F7E013058B0@sjhaexc02.sjha.org> I looked into purchasing this instrument. We decided that in order to satisfy the FDA requirements for formalin fixation, we would have to separate breasts or anything suspicious of breast mets and fix adequately in formalin before putting on this processor. We ended up not purchasing it, but I think this is an acceptable solution for that problem. Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rene J Buesa Sent: Wednesday, February 22, 2006 6:07 PM To: John PJ Coleman; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Microwave processing John: I know this instrument and the technology is safe. You will not need to do HIER for IHC since the tissue will be fixed in an alcohol based fixatives and I don't think that you will need to run parallel runs.Maybe you will have to dilute your antibodies a little more than for formalin + HIER. The turn around time is 120 cassettes per hour, and with 4 of these instruments (which I personally think is a "bit too much") what you will have to do is to rethink your whole staff scheduling. Try to get a book titled: Microwaves for the Art of Microscopy by L.P Kok and M.E.Boon, Coulomb Press Leyden 2003 (you can get it at http://www.coulomb.nl This instrument is ideal for a large volume reference lab like yours. Hope this will help you. Ren? J. John PJ Coleman wrote: I am the Senior tech of a large hospital corporation. My administration has just won funding for 4 Sakura Microwave rapid processing units. We run FISH her 2 on formalin fixed paraffin embedded tissue as per FDA protocol. As I tech, I am not in favor of tossing routine processing wholesale in favor of a completely new technology without thorough testing and parallel processing. Also, we are a regional reference lab for IHC and have a panel of 115 antibodies, all optimized for formalin fixed paraffin embedded tissue. We run an average of 350 IHC slides a day, max 580 per day and would have to re-optimize these to use in the new formalin free world while keeping our FFPE procedures in parallel for our reference lab work. Much like running 2 labs. If anyone has any insight, or if anyone currently uses these instruments for routine and/or IHC, feel free to call or email, and I'll check the postings on this string. We are also taking invitations to come out and see these things in use real time. John PJ Coleman-757 335-2159 http://members.cox.net/captainmadman/ http://journals.aol.com/jcolclefa/Waytoomuchboringcraptosayusually/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Brings words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From jqb7 <@t> cdc.gov Wed Feb 22 17:12:05 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Wed Feb 22 17:15:37 2006 Subject: [Histonet] Microwave processing References: Message-ID: We have the Sakura Xpress and we are still doing side-by-side comparisons on a variey of tissue types, IHC testing and special stains. So far the tissues form the Xpress have been as good as, and is some cases superior to, the traditional processing method. Jeanine Bartlett CDC Atlanta -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of John PJ Coleman Sent: Wed 2/22/2006 5:04 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Microwave processing I am the Senior tech of a large hospital corporation. My administration has just won funding for 4 Sakura Microwave rapid processing units. We run FISH her 2 on formalin fixed paraffin embedded tissue as per FDA protocol. As I tech, I am not in favor of tossing routine processing wholesale in favor of a completely new technology without thorough testing and parallel processing. Also, we are a regional reference lab for IHC and have a panel of 115 antibodies, all optimized for formalin fixed paraffin embedded tissue. We run an average of 350 IHC slides a day, max 580 per day and would have to re-optimize these to use in the new formalin free world while keeping our FFPE procedures in parallel for our reference lab work. Much like running 2 labs. If anyone has any insight, or if anyone currently uses these instruments for routine and/or IHC, feel free to call or email, and I'll check the postings on this string. We are also taking invitations to come out and see these things in use real time. John PJ Coleman-757 335-2159 http://members.cox.net/captainmadman/ http://journals.aol.com/jcolclefa/Waytoomuchboringcraptosayusually/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Wed Feb 22 17:15:45 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Wed Feb 22 17:15:53 2006 Subject: [Histonet] A call to action for US AP/Histologyprofessionals Message-ID: <83AACDB0810528418AA106F9AE9B7F7E013058B1@sjhaexc02.sjha.org> What a long sentence.. I sound like a pathologist! :>) Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Weems, Joyce Sent: Wednesday, February 22, 2006 6:06 PM To: Julia Dahl; Rcartun@harthosp.org; histonet@lists.utsouthwestern.edu; tkngflght@yahoo.com Subject: RE: [Histonet] A call to action for US AP/Histologyprofessionals Thanks for explaining it better. We really do need to work together on this and we need to make it happen before July 1 and not let it hang around like the cyto proficiency testing which took a year of frayed nerves and thousands of dollars before it got held. -----Original Message----- From: Julia Dahl [mailto:jdmd77@hotmail.com] Sent: Wednesday, February 22, 2006 6:01 PM To: Weems, Joyce; Rcartun@harthosp.org; histonet@lists.utsouthwestern.edu; tkngflght@yahoo.com Subject: RE: [Histonet] A call to action for US AP/Histologyprofessionals Thank you Joyce for your understanding of the issues at hand with the discrepant handling of MUEs for the people doing the procedure and those of us involved in making the diagnosis on the tissue procured from the procedure. If the MUE for a biopsy is TEN - then the MUE for the pathology services on biopsies should be TEN, not two. Yes, healthcare costs are rising and do need to be curtailed, blanket reduction in pathology services reimbursement out of proportion to other specialties - claiming that anything beyond two biopsies on a single patient on a single day are "unbelievable edits" so far outside the standard of care as to be considered equivalent to a typographical error is heinous. Every patient with chronic diarrhea, iron deficiency anemia, dysphagia, Barrett's esophagus, inflammatory bowel disease NEEDS to have more than two sites biopsied FOR the standard of care; every man with an elevated PSA, every woman with multiple breast lumps needs to have more than two sites biopsies FOR the standard of care to be maintained. Again, Joyce, thank you. The MUEs for pathology are not the answer to the current ails of the American Medical system. Julia Dahl, M.D. Pathologist >This is true, but it should be a fair thing. Why should a dermatologist >collect for 10 reimbursements and the pathologist only two of those ten? >j > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard >Cartun >Sent: Wednesday, February 22, 2006 4:13 PM >To: Histonet; Cheryl >Subject: Re: [Histonet] A call to action for US >AP/Histologyprofessionals > >One way or another, we must cut spending. Health care costs are out of >control (16% of our economic output is consumed by health care). Our >country cannot continue down this path. > >Richard > >Richard W. Cartun, Ph.D. >Director, Immunopathology & Histology >Assistant Director, Anatomic Pathology >Hartford Hospital >80 Seymour Street >Hartford, CT 06102 >(860) 545-1596 >(860) 545-0174 Fax > > >>> Cheryl 02/21/06 10:00AM >>> >Hello Folks-- > > There is another issue we should consider a call to paper the Senate >with letters stating our opinions....The AFIP faces closure due to >realignment by base closures (BRAC)--this includes the pathology center >AND the school!! We've lost so many good schools, and the AFIP is among >the best. There is a bill to relocate core services of the AFIP to >another center and the link below provides an easy way to write you your >senator in support of this bill: > > http://capwiz.com/ascpath/issues/alert/?alertid=8171231&type=CO > > Please take a minute to show your support for our science on MUE >billing (Peggy's email of earlier today),AFIP closure, and two >additional issues threatening continued viability of lab science in this >country. The form is easy to fill out, and you can forward the link to >several friends once you submit your letter (it will give you another >pop-up for sharing the letter once you click to submit yours) to >increase the impact of our unified voice. This link will lead you to >both letters and you can send it by email or snail mail: > http://capwiz.com/ascpath/home/ > > Thank you for your continued interest and support tp maintain >visibility and viability of our craft. Together we DO make a >difference. > > > >Cheryl Kerry, HT(ASCP) >Full Staff Inc. >Staffing the AP Lab by helping one Tech at a time. >281.883.7704 c >281.852.9457 o >admin@fullstaff.org > >Sign up for the FREE monthly newsletter AP News--updates, tricks of the >trade and current issues for Anatomic Pathology Clinical Labs. Send a >'subscribe' request to APNews@fullstaff.org. Please include your name >and specialty in the body of the email. >_______________________________________________ >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 > > >Confidentiality Notice ** The information contained in this message may be >privileged and is confidential information intended for the use of the >addressee listed above. If you are neither the intended recipient nor the >employee or agent responsible for delivering this message to the intended >recipient, you are hereby notified that any disclosure, copying, >distribution or the taking of any action in reliance on the contents of >this information is strictly prohibited. If you have received this >communication in error, please notify us immediately by replying to the >message and deleting it from your computer. Thank you. Saint Joseph's >Health System, Inc. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From jdmd77 <@t> hotmail.com Wed Feb 22 17:17:09 2006 From: jdmd77 <@t> hotmail.com (Julia Dahl) Date: Wed Feb 22 17:17:22 2006 Subject: [Histonet] A call to action for US AP/Histologyprofessionals In-Reply-To: <83AACDB0810528418AA106F9AE9B7F7E013058AF@sjhaexc02.sjha.org> Message-ID: >From the sources that I've been in touch with - the next week is actually the crucial time to interact with our specialty societies - specifically the ASCP and the CAP. The CAP is compiling a response to the MUEs and working with other specialty societies and the AMA to present a package of information to the legislators who will be able to effect a change in these edits. Waiting until July 1 will, unfortunately, be too late. If you have cases in which a patient's care would be substandard if only TWO biopsies were performed, those cases should be gathered, the patient's identifying information removed and those cases sent to the CAP (I will locate the contact information for anyone who needs it) for the summary of information to be worked in with the CAP/AMA/ASCP society approach to handling this. Julia Dahl, M.D. >From: "Weems, Joyce" >To: "Julia Dahl" >,,, >Subject: RE: [Histonet] A call to action for US AP/Histologyprofessionals >Date: Wed, 22 Feb 2006 18:06:09 -0500 >MIME-Version: 1.0 >Received: from sjhaowa01.sjha.org ([170.59.194.200]) by >bay0-mc3-f10.bay0.hotmail.com with Microsoft SMTPSVC(6.0.3790.211); Wed, 22 >Feb 2006 15:09:20 -0800 >Received: from sjhaowa01.sjha.org(170.59.194.200) by Webshield1000.sjha.org >via smtp id 767e_9db93632_a3f7_11da_9cab_0002b3e7320d;Wed, 22 Feb 2006 >18:04:46 -0500 >Received: from sjhaexc02.sjha.org ([10.2.0.18]) by sjhaowa01.sjha.org with >Microsoft SMTPSVC(6.0.3790.1830); Wed, 22 Feb 2006 18:06:09 -0500 >X-Message-Info: JGTYoYF78jFf6wazmum4to1PmFuBGHnAaipaHWtvJAk= >X-MimeOLE: Produced By Microsoft Exchange V6.5 >Content-class: urn:content-classes:message >X-MS-Has-Attach: X-MS-TNEF-Correlator: Thread-Topic: [Histonet] A call to >action for US AP/Histologyprofessionals >Thread-Index: AcY4A9QNZeYeXsnVR3OGUqlPm7X43gAAB58A >Return-Path: JWEEMS@sjha.org >X-OriginalArrivalTime: 22 Feb 2006 23:06:09.0912 (UTC) >FILETIME=[9185AF80:01C63804] >X-NAIMIME-Disclaimer: 1 >X-NAIMIME-Modified: 1 > >Thanks for explaining it better. We really do need to work together on >this and we need to make it happen before July 1 and not let it hang >around like the cyto proficiency testing which took a year of frayed >nerves and thousands of dollars before it got held. > >-----Original Message----- >From: Julia Dahl [mailto:jdmd77@hotmail.com] >Sent: Wednesday, February 22, 2006 6:01 PM >To: Weems, Joyce; Rcartun@harthosp.org; >histonet@lists.utsouthwestern.edu; tkngflght@yahoo.com >Subject: RE: [Histonet] A call to action for US >AP/Histologyprofessionals > > >Thank you Joyce for your understanding of the issues at hand with the >discrepant handling of MUEs for the people doing the procedure and those >of >us involved in making the diagnosis on the tissue procured from the >procedure. > >If the MUE for a biopsy is TEN - then the MUE for the pathology services >on >biopsies should be TEN, not two. > >Yes, healthcare costs are rising and do need to be curtailed, blanket >reduction in pathology services reimbursement out of proportion to other > >specialties - claiming that anything beyond two biopsies on a single >patient >on a single day are "unbelievable edits" so far outside the standard of >care >as to be considered equivalent to a typographical error is heinous. > >Every patient with chronic diarrhea, iron deficiency anemia, dysphagia, >Barrett's esophagus, inflammatory bowel disease NEEDS to have more than >two >sites biopsied FOR the standard of care; every man with an elevated >PSA, >every woman with multiple breast lumps needs to have more than two sites > >biopsies FOR the standard of care to be maintained. > >Again, Joyce, thank you. The MUEs for pathology are not the answer to >the >current ails of the American Medical system. > >Julia Dahl, M.D. >Pathologist > > > >This is true, but it should be a fair thing. Why should a dermatologist > >collect for 10 reimbursements and the pathologist only two of those >ten? > >j > > > >-----Original Message----- > >From: histonet-bounces@lists.utsouthwestern.edu > >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard > >Cartun > >Sent: Wednesday, February 22, 2006 4:13 PM > >To: Histonet; Cheryl > >Subject: Re: [Histonet] A call to action for US > >AP/Histologyprofessionals > > > >One way or another, we must cut spending. Health care costs are out of > >control (16% of our economic output is consumed by health care). Our > >country cannot continue down this path. > > > >Richard > > > >Richard W. Cartun, Ph.D. > >Director, Immunopathology & Histology > >Assistant Director, Anatomic Pathology > >Hartford Hospital > >80 Seymour Street > >Hartford, CT 06102 > >(860) 545-1596 > >(860) 545-0174 Fax > > > > >>> Cheryl 02/21/06 10:00AM >>> > >Hello Folks-- > > > > There is another issue we should consider a call to paper the Senate > >with letters stating our opinions....The AFIP faces closure due to > >realignment by base closures (BRAC)--this includes the pathology center > >AND the school!! We've lost so many good schools, and the AFIP is >among > >the best. There is a bill to relocate core services of the AFIP to > >another center and the link below provides an easy way to write you >your > >senator in support of this bill: > > > > http://capwiz.com/ascpath/issues/alert/?alertid=8171231&type=CO > > > > Please take a minute to show your support for our science on MUE > >billing (Peggy's email of earlier today),AFIP closure, and two > >additional issues threatening continued viability of lab science in >this > >country. The form is easy to fill out, and you can forward the link to > >several friends once you submit your letter (it will give you another > >pop-up for sharing the letter once you click to submit yours) to > >increase the impact of our unified voice. This link will lead you to > >both letters and you can send it by email or snail mail: > > http://capwiz.com/ascpath/home/ > > > > Thank you for your continued interest and support tp maintain > >visibility and viability of our craft. Together we DO make a > >difference. > > > > > > > >Cheryl Kerry, HT(ASCP) > >Full Staff Inc. > >Staffing the AP Lab by helping one Tech at a time. > >281.883.7704 c > >281.852.9457 o > >admin@fullstaff.org > > > >Sign up for the FREE monthly newsletter AP News--updates, tricks of the > >trade and current issues for Anatomic Pathology Clinical Labs. Send a > >'subscribe' request to APNews@fullstaff.org. Please include your name > >and specialty in the body of the email. > >_______________________________________________ > >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 > > > > > >Confidentiality Notice ** The information contained in this message may >be > >privileged and is confidential information intended for the use of the > >addressee listed above. If you are neither the intended recipient nor >the > >employee or agent responsible for delivering this message to the >intended > >recipient, you are hereby notified that any disclosure, copying, > >distribution or the taking of any action in reliance on the contents of > > >this information is strictly prohibited. If you have received this > >communication in error, please notify us immediately by replying to the > > >message and deleting it from your computer. Thank you. Saint Joseph's > >Health System, Inc. > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_________________________________________________________________ >Express yourself instantly with MSN Messenger! Download today - it's >FREE! >http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ > > > >Confidentiality Notice ** The information contained in this message may be >privileged and is confidential information intended for the use of the >addressee listed above. If you are neither the intended recipient nor the >employee or agent responsible for delivering this message to the intended >recipient, you are hereby notified that any disclosure, copying, >distribution or the taking of any action in reliance on the contents of >this information is strictly prohibited. If you have received this >communication in error, please notify us immediately by replying to the >message and deleting it from your computer. Thank you. Saint Joseph's >Health System, Inc. _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From laurie.reilly <@t> jcu.edu.au Wed Feb 22 17:33:19 2006 From: laurie.reilly <@t> jcu.edu.au (Laurie Reilly) Date: Wed Feb 22 17:33:22 2006 Subject: [Histonet] Re: Wright-Giemsa stain on tissue In-Reply-To: Message-ID: <5.1.0.14.0.20060223092436.00c4a608@mail.jcu.edu.au> Dear Dina and Histonetters, Giemsa stain on tissue sections was used some time ago looking for Erhlichia canis in dog tissues. The method was to make a very weak Giemsa solution and stain overnight. We used five drops of Giemsa in a Coplin jar of tap water ( ours is pH 5.5) Differentiate next day in very weak acetic acid. Regards, Laurie. At 01:07 PM 22/02/2006 -0600, Johnson, Teri wrote: >Dina, > >In my days in clinical labs, we did a May-Grunwald Giemsa stain on our >bone marrow biopsies and particle preps. I'm with Dr. Richmond in that >there the staining results from the Zenker's fixed material was second >to none. However, we were able to do a reasonably good job of it, >provided the pathologists never had to pull archival slides to compare. >That being said, I am happy to never go back to using it, what a pain! > >You might try using the M-G Giemsa technique and see how it works for >you. > >Teri Johnson, HT(ASCP)QIHC >Managing Director Histology Facility >Stowers Institute for Medical Research >1000 E. 50th St. >Kansas City, MO 64133 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mr.Laurie Reilly Ph 07 4781 4468 School of Veterinary & Biomedical Science Fax 07 4779 1526 James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. From JWEEMS <@t> sjha.org Wed Feb 22 17:36:11 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Wed Feb 22 17:36:19 2006 Subject: [Histonet] A call to action for US AP/Histologyprofessionals Message-ID: <83AACDB0810528418AA106F9AE9B7F7E013058B4@sjhaexc02.sjha.org> I have a copy of the top 100 tests to be affected, the ASCP letter and forms to complete if anyone is interested. Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: Julia Dahl [mailto:jdmd77@hotmail.com] Sent: Wednesday, February 22, 2006 6:17 PM To: Weems, Joyce; Rcartun@harthosp.org; histonet@lists.utsouthwestern.edu; tkngflght@yahoo.com Subject: RE: [Histonet] A call to action for US AP/Histologyprofessionals From the sources that I've been in touch with - the next week is actually the crucial time to interact with our specialty societies - specifically the ASCP and the CAP. The CAP is compiling a response to the MUEs and working with other specialty societies and the AMA to present a package of information to the legislators who will be able to effect a change in these edits. Waiting until July 1 will, unfortunately, be too late. If you have cases in which a patient's care would be substandard if only TWO biopsies were performed, those cases should be gathered, the patient's identifying information removed and those cases sent to the CAP (I will locate the contact information for anyone who needs it) for the summary of information to be worked in with the CAP/AMA/ASCP society approach to handling this. Julia Dahl, M.D. >From: "Weems, Joyce" >To: "Julia Dahl" >,,, >Subject: RE: [Histonet] A call to action for US AP/Histologyprofessionals >Date: Wed, 22 Feb 2006 18:06:09 -0500 >MIME-Version: 1.0 >Received: from sjhaowa01.sjha.org ([170.59.194.200]) by >bay0-mc3-f10.bay0.hotmail.com with Microsoft SMTPSVC(6.0.3790.211); Wed, 22 >Feb 2006 15:09:20 -0800 >Received: from sjhaowa01.sjha.org(170.59.194.200) by Webshield1000.sjha.org >via smtp id 767e_9db93632_a3f7_11da_9cab_0002b3e7320d;Wed, 22 Feb 2006 >18:04:46 -0500 >Received: from sjhaexc02.sjha.org ([10.2.0.18]) by sjhaowa01.sjha.org with >Microsoft SMTPSVC(6.0.3790.1830); Wed, 22 Feb 2006 18:06:09 -0500 >X-Message-Info: JGTYoYF78jFf6wazmum4to1PmFuBGHnAaipaHWtvJAk= >X-MimeOLE: Produced By Microsoft Exchange V6.5 >Content-class: urn:content-classes:message >X-MS-Has-Attach: X-MS-TNEF-Correlator: Thread-Topic: [Histonet] A call to >action for US AP/Histologyprofessionals >Thread-Index: AcY4A9QNZeYeXsnVR3OGUqlPm7X43gAAB58A >Return-Path: JWEEMS@sjha.org >X-OriginalArrivalTime: 22 Feb 2006 23:06:09.0912 (UTC) >FILETIME=[9185AF80:01C63804] >X-NAIMIME-Disclaimer: 1 >X-NAIMIME-Modified: 1 > >Thanks for explaining it better. We really do need to work together on >this and we need to make it happen before July 1 and not let it hang >around like the cyto proficiency testing which took a year of frayed >nerves and thousands of dollars before it got held. > >-----Original Message----- >From: Julia Dahl [mailto:jdmd77@hotmail.com] >Sent: Wednesday, February 22, 2006 6:01 PM >To: Weems, Joyce; Rcartun@harthosp.org; >histonet@lists.utsouthwestern.edu; tkngflght@yahoo.com >Subject: RE: [Histonet] A call to action for US >AP/Histologyprofessionals > > >Thank you Joyce for your understanding of the issues at hand with the >discrepant handling of MUEs for the people doing the procedure and those >of >us involved in making the diagnosis on the tissue procured from the >procedure. > >If the MUE for a biopsy is TEN - then the MUE for the pathology services >on >biopsies should be TEN, not two. > >Yes, healthcare costs are rising and do need to be curtailed, blanket >reduction in pathology services reimbursement out of proportion to other > >specialties - claiming that anything beyond two biopsies on a single >patient >on a single day are "unbelievable edits" so far outside the standard of >care >as to be considered equivalent to a typographical error is heinous. > >Every patient with chronic diarrhea, iron deficiency anemia, dysphagia, >Barrett's esophagus, inflammatory bowel disease NEEDS to have more than >two >sites biopsied FOR the standard of care; every man with an elevated >PSA, >every woman with multiple breast lumps needs to have more than two sites > >biopsies FOR the standard of care to be maintained. > >Again, Joyce, thank you. The MUEs for pathology are not the answer to >the >current ails of the American Medical system. > >Julia Dahl, M.D. >Pathologist > > > >This is true, but it should be a fair thing. Why should a dermatologist > >collect for 10 reimbursements and the pathologist only two of those >ten? > >j > > > >-----Original Message----- > >From: histonet-bounces@lists.utsouthwestern.edu > >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard > >Cartun > >Sent: Wednesday, February 22, 2006 4:13 PM > >To: Histonet; Cheryl > >Subject: Re: [Histonet] A call to action for US > >AP/Histologyprofessionals > > > >One way or another, we must cut spending. Health care costs are out of > >control (16% of our economic output is consumed by health care). Our > >country cannot continue down this path. > > > >Richard > > > >Richard W. Cartun, Ph.D. > >Director, Immunopathology & Histology > >Assistant Director, Anatomic Pathology > >Hartford Hospital > >80 Seymour Street > >Hartford, CT 06102 > >(860) 545-1596 > >(860) 545-0174 Fax > > > > >>> Cheryl 02/21/06 10:00AM >>> > >Hello Folks-- > > > > There is another issue we should consider a call to paper the Senate > >with letters stating our opinions....The AFIP faces closure due to > >realignment by base closures (BRAC)--this includes the pathology center > >AND the school!! We've lost so many good schools, and the AFIP is >among > >the best. There is a bill to relocate core services of the AFIP to > >another center and the link below provides an easy way to write you >your > >senator in support of this bill: > > > > http://capwiz.com/ascpath/issues/alert/?alertid=8171231&type=CO > > > > Please take a minute to show your support for our science on MUE > >billing (Peggy's email of earlier today),AFIP closure, and two > >additional issues threatening continued viability of lab science in >this > >country. The form is easy to fill out, and you can forward the link to > >several friends once you submit your letter (it will give you another > >pop-up for sharing the letter once you click to submit yours) to > >increase the impact of our unified voice. This link will lead you to > >both letters and you can send it by email or snail mail: > > http://capwiz.com/ascpath/home/ > > > > Thank you for your continued interest and support tp maintain > >visibility and viability of our craft. Together we DO make a > >difference. > > > > > > > >Cheryl Kerry, HT(ASCP) > >Full Staff Inc. > >Staffing the AP Lab by helping one Tech at a time. > >281.883.7704 c > >281.852.9457 o > >admin@fullstaff.org > > > >Sign up for the FREE monthly newsletter AP News--updates, tricks of the > >trade and current issues for Anatomic Pathology Clinical Labs. Send a > >'subscribe' request to APNews@fullstaff.org. Please include your name > >and specialty in the body of the email. > >_______________________________________________ > >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 > > > > > >Confidentiality Notice ** The information contained in this message may >be > >privileged and is confidential information intended for the use of the > >addressee listed above. If you are neither the intended recipient nor >the > >employee or agent responsible for delivering this message to the >intended > >recipient, you are hereby notified that any disclosure, copying, > >distribution or the taking of any action in reliance on the contents of > > >this information is strictly prohibited. If you have received this > >communication in error, please notify us immediately by replying to the > > >message and deleting it from your computer. Thank you. Saint Joseph's > >Health System, Inc. > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_________________________________________________________________ >Express yourself instantly with MSN Messenger! Download today - it's >FREE! >http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ > > > >Confidentiality Notice ** The information contained in this message may be >privileged and is confidential information intended for the use of the >addressee listed above. If you are neither the intended recipient nor the >employee or agent responsible for delivering this message to the intended >recipient, you are hereby notified that any disclosure, copying, >distribution or the taking of any action in reliance on the contents of >this information is strictly prohibited. If you have received this >communication in error, please notify us immediately by replying to the >message and deleting it from your computer. Thank you. Saint Joseph's >Health System, Inc. _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From TJJ <@t> Stowers-Institute.org Wed Feb 22 17:47:22 2006 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Wed Feb 22 17:47:50 2006 Subject: [Histonet] Mouse eye paraffin section Message-ID: What is the secret to good intact morphology in rodent/mouse eye paraffin processing? The retina detaches from the back of the eye, and there doesn't appear to be much we can do to keep it there. Is it because the vitreous humor is being replaced by fixative, alcohol, clearant, and paraffin? Does it help to inject fixative in the globe (quasi-perfuse)? Does it help to fix longer than 24 hours (I'm thinking probably so). What are others out there doing to keep your eye structures intact during paraffin processing? Thanks so much! Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From d.gianotti <@t> comcast.net Wed Feb 22 17:47:59 2006 From: d.gianotti <@t> comcast.net (Dennis Gianotti) Date: Wed Feb 22 17:48:07 2006 Subject: [Histonet] penfix Message-ID: <43FCF82F.9070800@comcast.net> Is there another fixative that might be better for fatty breasts besides penfix? We currently use penfix, but we are looking for better options if they are out there. We fix the cassettes of breast tissue in penfix for 3 hours before we put them in formalin and onto the processors. Occasionally, we still have to reprocess breast tissue and we are trying to avoid this. Yes, the PA's are trying to cut thin sections and not pack the cassettes. We process about 300 breast cassettes or more per day and try to spread them between 3 VIP processors amid our other gyne and placenta tissues. thanks for any suggestions From bhewlett <@t> cogeco.ca Wed Feb 22 18:05:39 2006 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Wed Feb 22 18:05:48 2006 Subject: [Histonet] Microwave processing References: <20060222230636.28591.qmail@web61219.mail.yahoo.com> Message-ID: <000701c6380c$e1b0c820$6400a8c0@mainbox> Rene, and John, The answer for use of this instrument for routine IHC is to make sure that fixation PRECEDES processing, they are not one and the same! If your routine fixation is NBF, then prefix in NBF before rapid processing. True, some of the speed advantages are lost, but rapid processing can still be used to good effect in the workflow. Any change in the routine fixative used can, and does, have dramatic effects on the antigen to be demonstrated by IHC. The effects on antigens such as CK's, vimentin etc. can be positive, as Rene indicates no HIER is required. On the other hand, the effects on ER, HER2 and many other surface proteins can be devastating! There is a reason that the fixation protocol for HER2, EGFr and other surface markers calls for fixation in NBF, i.e. many surface proteins are known to be stripped away by alcohol fixation!! FISH for HER2 is not a problem with alcohol fixation, except that the proteolysis step will have to be removed or greatly curtailed. This means you will no longer be following the approved methodology!!! IMHO, to not parallel process and compare/validate all results would constitute a gross dereliction of duty!!! Bryan ----- Original Message ----- From: "Rene J Buesa" To: "John PJ Coleman" ; Sent: Wednesday, February 22, 2006 6:06 PM Subject: Re: [Histonet] Microwave processing > John: > I know this instrument and the technology is safe. You will not need to > do HIER for IHC since the tissue will be fixed in an alcohol based > fixatives and I don't think that you will need to run parallel runs.Maybe > you will have to dilute your antibodies a little more than for formalin + > HIER. > The turn around time is 120 cassettes per hour, and with 4 of these > instruments (which I personally think is a "bit too much") what you will > have to do is to rethink your whole staff scheduling. > Try to get a book titled: > Microwaves for the Art of Microscopy by L.P Kok and M.E.Boon, Coulomb > Press Leyden 2003 (you can get it at http://www.coulomb.nl > This instrument is ideal for a large volume reference lab like yours. > Hope this will help you. > Ren? J. > > John PJ Coleman wrote: > I am the Senior tech of a large hospital corporation. My > administration has just won funding for 4 Sakura Microwave rapid > processing units. We run FISH her 2 on formalin fixed paraffin embedded > tissue as per FDA protocol. As I tech, I am not in favor of tossing > routine processing wholesale in favor of a completely new technology > without thorough testing and parallel processing. Also, we are a > regional reference lab for IHC and have a panel of 115 antibodies, all > optimized for formalin fixed paraffin embedded tissue. We run an > average of 350 IHC slides a day, max 580 per day and would have to > re-optimize these to use in the new formalin free world while keeping > our FFPE procedures in parallel for our reference lab work. Much like > running 2 labs. If anyone has any insight, or if anyone currently uses > these instruments for routine and/or IHC, feel free to call or email, > and I'll check the postings on this string. We are also taking > invitations to come out and see these things in use real time. > > John PJ Coleman-757 335-2159 > http://members.cox.net/captainmadman/ > http://journals.aol.com/jcolclefa/Waytoomuchboringcraptosayusually/ > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > --------------------------------- > Brings words and photos together (easily) with > PhotoMail - it's free and works with Yahoo! Mail. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From kappeler <@t> patho.unibe.ch Thu Feb 23 00:26:21 2006 From: kappeler <@t> patho.unibe.ch (Andi Kappeler) Date: Thu Feb 23 00:27:08 2006 Subject: [Histonet] PGP 9.5 References: <001b01c63747$7ede4760$ede49440@HPPav2> Message-ID: <00c901c63842$240915d0$27955c82@patho.unibe.ch> Hi Peggy we use mo-a-PGP 9.5, clone 13C4, Biomeda Cat Nr. V3231. Working conc. is 1.5 ug Ig/ml (corresponding to 1:200), pretreatment HIER (microwave) citrate, on FFPE tissue, cut at 2-3 um. Working conc. will depend on visualization system used. In our lab we visualize with StrABC method (either with peroxidase - DAB or alkaline phosphatase - new fuchsin, depending on specificproject). Hope this helps. Andi Kappeler Institute of Pathology, University of Bern, Switzerland ----- Original Message ----- From: "Lee & Peggy Wenk" To: Sent: Wednesday, February 22, 2006 1:32 AM Subject: [Histonet] PGP 9.5 > Anyone have experience with PGP 9.5 (Protein Gene Product)? It > demonstrates > nerve fibers. One of our pathologists wants to use it on skin biopsies for > diagnosis of neuropathies. > > We would prefer peroxidase for paraffin sections, rather than FITC or FS. > Just wondering if people are having better luck with monoclonals or > polyclonals, and if there is a difference in animal species for the > primary > (we'll be doing this on human skin). > > Also - all the papers we find mention thick sections and using a confocal > microscope. Anyone doing PGP on 5 um sections and/or with a regular light > microscope? > > One of my students is doing this as her research project. She's done a lot > of reading on it, and most papers don't discuss the actual procedure or > the > type of antibody, etc. They are on the research aspect, or the diagnostic > aspect, not on what histotechs need to do the procedures. Those articles > that do mention techniques have very different methods (of course), and > are > missing important information needed by histotechs. > > We would appreciate a couple of little hints of what works for "real life > histotechs", to get us started in the right direction for use in a routine > clinical immunohistochemistry lab. > > Feel free to contact me directly. Vendors included. I'll summarize and > relay > the info back to Histonet. > > Thanks in advance. > > Peggy A. Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From d.fuster <@t> ub.edu Thu Feb 23 06:18:37 2006 From: d.fuster <@t> ub.edu (Dolors Fuster) Date: Thu Feb 23 06:18:56 2006 Subject: [Histonet] Help with immunofluorescence Message-ID: <43FDA81D.30404@ub.edu> We are trying to demonstrate SHH and Survivin in human embrionic colo-rectal tissue by Immunofluorescence in FFPE samples. We are using: * goat polyclonal SHH (N-19) sc-1194 with donkey anti-goat secondary sc-2024 * mouse monoclonal Survivin (D-8) sc-17779 with donkey anti-mouse secondary sc-2099 As antigen retrieval we have used saponin 0,1% in PBS in some slides and a post-fixation in carnoy in others. Carnoy postfixed slides seem to shoe a better image. However,and specially in Sonic, we are seeing that some cells in the submucosa appear labeled while epitelial cells in the mucosa remain unlabeled As we are seing these images in samples treated only with the conjugated Ab but also in samples with primary and conjugated, we think it could be a cross or unespecific reaction of the secondary Ab or may be the primary has not reacted properly. May you help us at this point? We would also ask you for some suggestion about tissue controls for these Ab. Any advice would be appreciated. Thanks in advance Dolors Fuster T?cnic Especialista en Anatomia Patol?gica i Citologia Departament d'Anatomia i Embriologia Humana Facultat de Medicina (HCP) Universitat de Barcelona From jqb7 <@t> cdc.gov Thu Feb 23 06:26:16 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Thu Feb 23 06:31:50 2006 Subject: [Histonet] disposal of xylol waste from CJD cases Message-ID: Is this xylene that is from the stainer or the processor or both? And is it generated from tissues that have been first formic acid-treated? Jeanine Bartlett, BS, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sharon Allen Sent: Wednesday, February 22, 2006 3:20 PM To: Histonet (E-mail) Subject: [Histonet] disposal of xylol waste from CJD cases I am having a problem disposing or our xylol waste from Prion testing for CJD. Our waste disposal company has just informed us thay they will take all other waste but won't touch the xylol (it has all been treated with javex). Apparently they have been storing it all these years & now are refusing to accept anymore. I would like to know how other labs testing for CJD are disposing of their xylol. Does anyone use a xylol substitute? Any ideas would be very helpful. Right now I have it packaged & sitting on the floor in the lab which goes against all the safety rules. Sharon sallen@hsc.mb.ca This e-mail and/or any documents in this transmission is intended for the address(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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jennifer.l.hofecker <@t> Vanderbilt.Edu Thu Feb 23 08:23:16 2006 From: jennifer.l.hofecker <@t> Vanderbilt.Edu (Hofecker, Jennifer L) Date: Thu Feb 23 08:23:25 2006 Subject: [Histonet] Tennessee meeting program finalized Message-ID: <898D946569A27444B65667A49C074052852BAF@mailbe06.mc.vanderbilt.edu> Good Morning Histonetters. The meeting programs have been finalized and mailed out for our annual meeting. It will be held June 1-3 in Chattanooga. We will offer both traditional 3 hour workshops as well as 1.5 hour lectures. All educational offerings will be held on Friday and Saturday from 8:00 to 4:30. Workshops and Lectures to be presented include: Equipment Care and Maintenance Specimen Tracking Quality Control an Stanardization of IHC Sectioning Artifacts: Causes and Cures Double Immunostaining Wet Workshop Control Blocks and Tissues for Histology and Immunology Laboratory Management Problems and Solutions Lymphomas, Reward and Recognition a Toolbox for success Creutzfeldt - Jakob Disease and the Prion Disorders: Myths, Facts and Mysteries Diagnosing Hirschsprung's Disease: A leveling experience We have planned a river boat cruise for Friday night and we have an excellent group of presenters. We also have a great rate on our rooms this year! If you didn't get your program or if you'd like more information, please contact me directly. The deadline to take advantage of our hotel room rate is May 1. Thanks and have a great day, Jennifer Hofecker, TSH President Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax From akbitting <@t> geisinger.edu Thu Feb 23 10:06:23 2006 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Thu Feb 23 10:06:49 2006 Subject: [Histonet] Materials retention Message-ID: Does anyone have info about the PA Dept. of Health's slide and block retention requirements? IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From RBARNHART <@t> summithealth.org Thu Feb 23 10:30:35 2006 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Thu Feb 23 10:30:56 2006 Subject: [Histonet] Laugh for the day Message-ID: I knew all the histonetters would enjoy this one. I know I haven't been able to stop chuckling to myself since. We have 11th and 12th grade students from the high school come to our hospital and go through each of the departments to expose them to the different jobs in the hospital. Lately a lot of the students have been extremely interested in histology which is great. The student today was priceless. After I went through explaining all that we do, the student made the comment "Oh it looks easy". I said "really?". Her response "Oh yeah this is stuff I learned in 6th grade". I explained that this is much more then a 6th grade education. Her attitude was not directed at histology alone but the entire lab. You have to love teenagers. From jqb7 <@t> cdc.gov Thu Feb 23 10:28:34 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Thu Feb 23 10:39:00 2006 Subject: [Histonet] paraffin repellent Message-ID: Hi everyone: A while back there was a post about a product that was similar to Para Gard except it didn't leave as much residue. Can anyone help me out with the name of the product? Thanks, Jeanine Bartlett, BS, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov From nadams <@t> CapeCodHealth.org Thu Feb 23 12:30:47 2006 From: nadams <@t> CapeCodHealth.org (Adams, Nancy) Date: Thu Feb 23 12:30:59 2006 Subject: [Histonet] contact information for VWR Message-ID: <17A1862099540D458C8FE9380C2BC461C0933C@fh2xmail.fhdomain1.capecodhealth.org> Looking for contact information for VWR supplies. Thanks Nancy R. ************************************************************ This email and any files transmitted with it are confidential. And intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please contact the system administrator for Cape Cod Healthcare. Administrator@CapeCodHealth.org ************************************************************ From KevinMcGovern <@t> catholichealth.net Thu Feb 23 12:36:31 2006 From: KevinMcGovern <@t> catholichealth.net (McGovern, Kevin) Date: Thu Feb 23 12:36:46 2006 Subject: [Histonet] contact information for VWR Message-ID: I can help you with that: http://www.vwrsp.com/ Kev -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Adams, Nancy Sent: Thursday, February 23, 2006 12:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] contact information for VWR Looking for contact information for VWR supplies. Thanks Nancy R. ************************************************************ This email and any files transmitted with it are confidential. And intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please contact the system administrator for Cape Cod Healthcare. Administrator@CapeCodHealth.org ************************************************************ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SHARON.OSBORN <@t> SPCORP.COM Wed Feb 8 14:06:03 2006 From: SHARON.OSBORN <@t> SPCORP.COM (Osborn, Sharon) Date: Thu Feb 23 12:52:08 2006 Subject: [Histonet] Slide Printers Message-ID: <29B25753F6B1D51196110002A589D444032C4A03@PALMSG30.us.schp.com> Rebecca, Several months ago, I did a post about the Leica printers (not labelers). Both the Leica and Sakura were developed through a joint partnership agreement in the beginning so they began life as the same printers with different company names and different software operation. Leica custom made their software while Sakura used an off the shelf software for operation of the printers. I do not know about Sakura in the past year or their contract about the upgrades, etc. so I canot speak any further on that. However, Leica has listened to the customer and made tremendous improvements, upgrades, etc for better operation of the printers. We originally had the TBS etch slides and burn labels cassettes. It was the best of its time and for years, the only item out there. However, the Leica is wonderful! The service is responsive in answering questions or doing needed work. I encourage you to check closely into whether Sakura has kept up with all the improvements and upgrades. I can whole heartedly recommend the Leica printers for ease of use, speed, cleanliness (NO DUST on the slides that has to be washed off!) These are printers because they use a patented technology of a printer with special ink that is cured with a UV light before released for our use. They are clear to read, fairly quiet and operator friendly. Our researchers and post docs have been trained to use the equipment(they also used the TBS). When these came, they all but brought us flowers for making the work so much easier and cleaner. sharon osborn histology DNAX, SP BioPharma Palo Alto, CA 650.496.6539 Date: Tue, 7 Feb 2006 13:48:22 -0500 From: "LeVier, Rebecca J" Subject: [Histonet] Cassette Labeling To: Message-ID: <0FDBF29200C637468CCC1F9E7E85A3D214E8FE@MAIL-LR.lha.org> Content-Type: text/plain; charset="iso-8859-1" Could anyone give me some feedback on the Leica Cassette Labeler or the Sakura Cassette Labeler? Thanks ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From dsantana <@t> pmaonline.com Thu Feb 23 13:11:05 2006 From: dsantana <@t> pmaonline.com (Santana, Diane) Date: Thu Feb 23 13:10:55 2006 Subject: [Histonet] bx sponges VS no bx sponges Message-ID: <4C96AA62BADFD81180CB0002A5AD67FB0711388B@MAILPMA> I have a question to put out there.... In the past I have had a problem with dry artifact on my bx tissue. I finally straighten it out with the help of all you histo netters. Yet every so often it comes up again, only 1 or 2 cases. (compared to LOTS) Anyway for the last several weeks, I have had no problems....none. Than yesterday I could tell I was going to have problems as I was cutting. Now here's the question...... For the last couple weeks, one of my pathologist has been gone, now he is back and when he gross's he never uses the blue bx sponges. When my other pathologist gross, she always uses them. The only difference from the weeks past and yesterday is no bx sponges. Can this really be the problem? I have a hard time thinking it, but its the only difference in the times of problems and not. What do you guys think? Diane Santana PMA Haverhill, Mass From Jackie.O'Connor <@t> abbott.com Thu Feb 23 14:10:12 2006 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Thu Feb 23 14:10:41 2006 Subject: [Histonet] Melanin and Azure B Message-ID: Hi - I found a couple of old posts about being able to stain melanin with 1% aqueous Azure B. Can someone shed light on why this isn't seeming to work for me? I'm basically trying to identify a black pigment in FFPE that looks like melanin without having to depend on HMB45. Any takers? Jackie From failm <@t> musc.edu Thu Feb 23 14:25:40 2006 From: failm <@t> musc.edu (Mildred Fail) Date: Thu Feb 23 14:26:16 2006 Subject: [Histonet] Melanin and Azure B Message-ID: Jackie, you can put the slidesin30% hydrogen peroxide overnight this should bleach out melanin Rena Rena Fail >>> "Jackie M O'Connor" 02/23/06 03:10PM >>> Hi - I found a couple of old posts about being able to stain melanin with 1% aqueous Azure B. Can someone shed light on why this isn't seeming to work for me? I'm basically trying to identify a black pigment in FFPE that looks like melanin without having to depend on HMB45. Any takers? Jackie _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Thu Feb 23 14:47:34 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Thu Feb 23 14:47:45 2006 Subject: [Histonet] Melanin and Azure B References: Message-ID: <43FE1F66.2D8C17FB@uwo.ca> If the pigment is already black, it doesn't need to be stained. Melanin is bleached by 3% hydrogen peroxide ("10 volumes available oxygen"), 3 to 36 hours at room temperature. (Control slide in water.) Bleaching by H2O2 is considered a reasonably specific test for melanin. Lillie & Fullmer (p.524) comment that melanin is stained dark green by 0.05% azure A at pH 1 for 20 min. This coloration resisted methylation, so it may be due to oxidized cystine in an associated protein. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Jackie M O'Connor wrote: > > Hi - > I found a couple of old posts about being able to stain melanin with 1% > aqueous Azure B. Can someone shed light on why this isn't seeming to work > for me? > I'm basically trying to identify a black pigment in FFPE that looks like > melanin without having to depend on HMB45. Any takers? > > Jackie > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From amosbrooks <@t> gmail.com Thu Feb 23 15:41:12 2006 From: amosbrooks <@t> gmail.com (Amos Brooks) Date: Thu Feb 23 15:41:26 2006 Subject: [Histonet] Microwwave Processing: Fixation vs Standardization Message-ID: <582736990602231341s5dbbbcc2je3d87933b80017fa@mail.gmail.com> Greg, I would like to hear of your experiences with this as well. I am seeing an ugly situation brewing on the horizon with fixation and standardization. With the introduction of certain tests that are directly related to diagnosis and treatment such as FISH and Herceptest as well as other tests that are highly standardized we are going to have to be very conscious of fixation and it's affect on the end results of the test. My opinion is that it would be preferred to have a standard fixation and I truly do not care which one it is as long as it is standardized. Maintaining multiple procedures seems to be not only labor intensive but could be nearly impossible due in part to the fact that multiple procedures would require multiple controls. Some controls are very difficult to find such as Varicella Virus, Alk-1 or even HSV sometimes to name a few. I really see so many problems with this that I can't see any way around it and it is something that we will all end up having to deal with eventually since these tests are quickly becoming standard and who's to say which tests are going to be needed prior to processing. Thanks, Amos Brooks Message: 16 Date: Wed, 22 Feb 2006 14:28:17 -0800 From: "Luck, Greg D." Subject: RE: [Histonet] Microwave processing To: 'John PJ Coleman' , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain John, Your concerns are very valid particularly in your role as a reference lab. A couple of questions. 1st, which part of "FFPE" does the Sakura instrument not fulfill (e.g. are the tissues not exposed to NBF during the Sakura processing cycle). 2nd, When you perform IHC in the role as a reference lab how are you currently ascertaining how the paraffin blocks you receive for IHC have been processed? In a somewhat analogous situation for years we maintained two IHC protocols because we did histology for two hospitals. One hospital used NBF as its designated routine fixative and the other used "Prefer". Was somewhat of a pain but was manageable, however we only average about 50-60 IHC's/day. I could share some of what we learned developing and maintaining two standing IHC protocols for each antibody if you think our situation has any potential learning value for you. Good luck, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org www.deaconessmedicalcenter.org From Linresearch <@t> aol.com Thu Feb 23 16:31:31 2006 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Thu Feb 23 16:31:46 2006 Subject: [Histonet] Bar code system Message-ID: <19a.461e6001.312f91c3@aol.com> Hi, Is there a reliable bar code system that any one is using to lable specimen containers, slides and labels in Histology especially on the pharmaceutical side? Linda From Lobke.DeBels <@t> UGent.be Fri Feb 24 01:58:06 2006 From: Lobke.DeBels <@t> UGent.be (Lobke De Bels) Date: Fri Feb 24 01:58:19 2006 Subject: [Histonet] staining collagen Message-ID: <1140767886.43febc8e333ac@mail.ugent.be> Von Gieson gives a good contrast for the human eye, but the computer can't make a difference between yellow/orange and red. If I only use acid fushine (or aniline) everything stains pink(blue). I have to find a way to stain the connective tissue so the computer can clearly make a difference between connective tissue and the other tissue. The ideal should be a method that only stains collagen. Many thanks in advance for any help Lobke De Bels Department of Morphology Faculty of Veterinary Medicine Ghent University Salisburylaan 133 9820 Merelbeke Belgium lobke.debels@ugent.be -- From barbara.bublava <@t> meduniwien.ac.at Fri Feb 24 02:15:59 2006 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Fri Feb 24 02:17:02 2006 Subject: [Histonet] staining collagen References: <1140767886.43febc8e333ac@mail.ugent.be> Message-ID: <006701c6391a$8c899e80$47a9abc1@GERICHTS9XOZZ8> You could use anilineblue instead of acid fuchsin. makes better contrast and is described in John Kiernans book tell me if you can?t find a protocol greetings Barbara, Vienna ----- Original Message ----- From: "Lobke De Bels" To: Sent: Friday, February 24, 2006 8:58 AM Subject: [Histonet] staining collagen Von Gieson gives a good contrast for the human eye, but the computer can't make a difference between yellow/orange and red. If I only use acid fushine (or aniline) everything stains pink(blue). I have to find a way to stain the connective tissue so the computer can clearly make a difference between connective tissue and the other tissue. The ideal should be a method that only stains collagen. Many thanks in advance for any help Lobke De Bels Department of Morphology Faculty of Veterinary Medicine Ghent University Salisburylaan 133 9820 Merelbeke Belgium lobke.debels@ugent.be -- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo.rogerson <@t> waht.swest.nhs.uk Fri Feb 24 02:43:01 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Fri Feb 24 02:43:16 2006 Subject: [Histonet] Laugh for the day Message-ID: That can't be true; well it's certainly not true in the UK as we all know that all the Lab disciplines are taught in the first semester at Medical School and all graduates are able to work seamlessly in Labs with no further training. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 "One of the benefits of getting older is finding wisdom but so many get of us get older and never find it; sadly that is a waste of a life." -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: Thursday, February 23, 2006 4:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Laugh for the day I knew all the histonetters would enjoy this one. I know I haven't been able to stop chuckling to myself since. We have 11th and 12th grade students from the high school come to our hospital and go through each of the departments to expose them to the different jobs in the hospital. Lately a lot of the students have been extremely interested in histology which is great. The student today was priceless. After I went through explaining all that we do, the student made the comment "Oh it looks easy". I said "really?". Her response "Oh yeah this is stuff I learned in 6th grade". I explained that this is much more then a 6th grade education. Her attitude was not directed at histology alone but the entire lab. You have to love teenagers. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Fri Feb 24 07:41:56 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Feb 24 07:42:04 2006 Subject: [Histonet] staining collagen In-Reply-To: <1140767886.43febc8e333ac@mail.ugent.be> Message-ID: <20060224134156.37206.qmail@web61214.mail.yahoo.com> Lobke: Why don't you try using some colour complementary filters between the light source and the slide? Ren? J. Lobke De Bels wrote: Von Gieson gives a good contrast for the human eye, but the computer can't make a difference between yellow/orange and red. If I only use acid fushine (or aniline) everything stains pink(blue). I have to find a way to stain the connective tissue so the computer can clearly make a difference between connective tissue and the other tissue. The ideal should be a method that only stains collagen. Many thanks in advance for any help Lobke De Bels Department of Morphology Faculty of Veterinary Medicine Ghent University Salisburylaan 133 9820 Merelbeke Belgium lobke.debels@ugent.be -- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Brings words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. From Janet.Bonner <@t> FLHOSP.ORG Fri Feb 24 09:37:26 2006 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Feb 24 09:40:33 2006 Subject: [Histonet] Laugh for the day Message-ID: Actually, it's not true (what she said), she just didn't grasp the extent of what she was seeing..It's often quite a compliment to hear this, knowing we are so good at what we do that we make it look easy!!!! Janet ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Kemlo Rogerson Sent: Fri 2/24/2006 3:43 AM To: 'Rebecca Barnhart'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Laugh for the day That can't be true; well it's certainly not true in the UK as we all know that all the Lab disciplines are taught in the first semester at Medical School and all graduates are able to work seamlessly in Labs with no further training. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 "One of the benefits of getting older is finding wisdom but so many get of us get older and never find it; sadly that is a waste of a life." -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: Thursday, February 23, 2006 4:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Laugh for the day I knew all the histonetters would enjoy this one. I know I haven't been able to stop chuckling to myself since. We have 11th and 12th grade students from the high school come to our hospital and go through each of the departments to expose them to the different jobs in the hospital. Lately a lot of the students have been extremely interested in histology which is great. The student today was priceless. After I went through explaining all that we do, the student made the comment "Oh it looks easy". I said "really?". Her response "Oh yeah this is stuff I learned in 6th grade". I explained that this is much more then a 6th grade education. Her attitude was not directed at histology alone but the entire lab. You have to love teenagers. _______________________________________________ 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 ------------------------------- The information contained in this message may be privileged and / or confidential and protected from disclosure. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From c.ingles <@t> hosp.wisc.edu Fri Feb 24 09:46:49 2006 From: c.ingles <@t> hosp.wisc.edu (Ingles Claire) Date: Fri Feb 24 09:50:41 2006 Subject: [Histonet] Laugh for the day References: Message-ID: <583D3E9A1E843445BD54E461D1A2F6F317A04F11@uwhis-xchng2.hosp.wisc.edu> Yet kind of scary. Highlighting the fact once again that our talents are usually taken for granted and that anyone can do it! Yes, it does give me a warm feeling that we are able to do such a difficult, multi-faceted job and make it look much easier than it is. Let's hope some of those kids are steered in our direction in a couple of years! Claire ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bonner, Janet Sent: Fri 2/24/2006 9:37 AM To: Kemlo Rogerson; Rebecca Barnhart; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Laugh for the day Actually, it's not true (what she said), she just didn't grasp the extent of what she was seeing..It's often quite a compliment to hear this, knowing we are so good at what we do that we make it look easy!!!! Janet ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Kemlo Rogerson Sent: Fri 2/24/2006 3:43 AM To: 'Rebecca Barnhart'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Laugh for the day That can't be true; well it's certainly not true in the UK as we all know that all the Lab disciplines are taught in the first semester at Medical School and all graduates are able to work seamlessly in Labs with no further training. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 "One of the benefits of getting older is finding wisdom but so many get of us get older and never find it; sadly that is a waste of a life." -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: Thursday, February 23, 2006 4:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Laugh for the day I knew all the histonetters would enjoy this one. I know I haven't been able to stop chuckling to myself since. We have 11th and 12th grade students from the high school come to our hospital and go through each of the departments to expose them to the different jobs in the hospital. Lately a lot of the students have been extremely interested in histology which is great. The student today was priceless. After I went through explaining all that we do, the student made the comment "Oh it looks easy". I said "really?". Her response "Oh yeah this is stuff I learned in 6th grade". I explained that this is much more then a 6th grade education. Her attitude was not directed at histology alone but the entire lab. You have to love teenagers. _______________________________________________ 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 ------------------------------- The information contained in this message may be privileged and / or confidential and protected from disclosure. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From pathrm35 <@t> adelphia.net Fri Feb 24 10:57:36 2006 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Feb 24 10:57:44 2006 Subject: [Histonet] Medical Director job description Message-ID: <24435074.1140800256954.JavaMail.root@web21> Fellow techs, Is anyone willing to share the following? I need a job description for a Medical Director for high complexity testing in a dermpath lab. Thanks, Ron Martin fax 561-721-1249 From jlinda <@t> ces.clemson.edu Fri Feb 24 13:22:33 2006 From: jlinda <@t> ces.clemson.edu (Linda Jenkins) Date: Fri Feb 24 13:18:31 2006 Subject: [Histonet] Region III Registration Deadline Approaching Message-ID: <5.2.1.1.2.20060224141714.01f736e0@mailhost.ces.clemson.edu> The Region III registration deadline is March 1st. Make sure you get your registration turned in to Debbie Ellenburg at DEllenburg2@stfrancishealth.org. Don't miss the boat! We need an accurate head count for our "3 hour tour" and dinner cruise. Hope to see you there, Linda Linda Jenkins, HT Clemson University Dept. of Bioengineering Clemson, SC 29634-0905 864.656.5553 http://www.ces.clemson.edu/bio/research/histo.htm From galinadeyneko <@t> yahoo.com Fri Feb 24 13:34:58 2006 From: galinadeyneko <@t> yahoo.com (Galina Deyneko) Date: Fri Feb 24 13:35:06 2006 Subject: [Histonet] anti-rabbit alpha-actin Message-ID: <20060224193458.86786.qmail@web33111.mail.mud.yahoo.com> Dear colleagues. Could you please recommend anti-alpha-actin smooth muscle cell antibody for rabbit aorta (formalin fixed paraffin embedded). I found several from different companies, but it is not specific for rabbit. Any prompts (dilution, retrieval,detection) would be highly appreciated. Galina Deyneko Novartis Cambridge MA 617-871-7613 --------------------------------- Yahoo! Mail Use Photomail to share photos without annoying attachments. From Linke_Noelle <@t> Allergan.com Fri Feb 24 13:44:44 2006 From: Linke_Noelle <@t> Allergan.com (Linke_Noelle) Date: Fri Feb 24 13:45:14 2006 Subject: [Histonet] anti-rabbit alpha-actin Message-ID: LabVision's works nicely on rabbit (data sheet specifies rabbit as well so you can return it if there's a problem), no retrieval needed. Noelle Linke, BS, HTL(ASCP)QIHC Allergan, Inc 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5568 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Galina Deyneko Sent: Friday, February 24, 2006 11:35 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] anti-rabbit alpha-actin Dear colleagues. Could you please recommend anti-alpha-actin smooth muscle cell antibody for rabbit aorta (formalin fixed paraffin embedded). I found several from different companies, but it is not specific for rabbit. Any prompts (dilution, retrieval,detection) would be highly appreciated. Galina Deyneko Novartis Cambridge MA 617-871-7613 --------------------------------- Yahoo! Mail Use Photomail to share photos without annoying attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Fri Feb 24 14:25:52 2006 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Feb 24 14:35:43 2006 Subject: [Histonet] Re: Do you Love your job? Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D666@usca0082k08.labvision.apogent.com> Peggy, I do love the histology profession! I started in electron microscopy and did that for over a decade, but as I was in a hospital lab, I ended up helping out in the histology lab (never saw one before that - at least in a hospital. I ended up leaving EM, mainly due to dearth of jobs in that field, and have done everything from routine histology to histochemistry, immunochemistry ISH, imaging, etc. I now work for a company that sells antibodies and manufactures lab instruments, including an immunostainer. The sky is the only limit to what you can do and where you can go in this profession. Histonet gives a great example that there is an extraordinary breadth of expericences to be had in this field. We are in great demand, and we are limited only by our imagination. I thought I had done it all in the lab -from diagnostics to research, but then I got into the business side of it and a whole new world opened up - now instead of wishing "someone would make such-and-such" I actually can get it made! And then I travel around the country, indeed, the world, to help people learn to do this work! One thing I learned is that you have to move around to advance yourself. Most labs are too small to support unending advancement. Therefore, you must learn whatever there is to learn in a particular lab - forget about a job description - do anything there is to do. That will broaden your horizons and open up lots of opportunities. Then, when the job gets routine, and you have reached the limit of what you can do there, move on and find a new challenge. Good luck. I know you will have fun with it. Tim Morken I am a second semester student at Argosy University in the HT program. I was wondering what the pros and cons are in the Histonet world. I have been in the service industry in one form or another for the last 30 years. This is a total diversion from my past life and I would like to know what you love about your job and what you hate about your job. If anyone would like to give me some input, I would greatly appreciate it. Thanks, Peggy _______________________________________________ Histonet mailing list Tim Morken Product Development Lab Vision - Neomarkers 47777 Warm Springs Blvd. Fremont, CA 94539 USA tel: 01-510-991-2840 fax: 01 510-991-2826 email: tpmorken@labvision.com web: www.labvision.com Free webhosting for US State Histotechnology Societies: http://www.labvisioncorp.com/demowebsite/index.cfm From Barry.R.Rittman <@t> uth.tmc.edu Fri Feb 24 14:47:20 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Fri Feb 24 14:47:23 2006 Subject: [Histonet] Re: Do you Love your job? Message-ID: Unfortunately I do not get to carry out a lot of bench procedures anymore. I did and do love histological techniques as you always see the end result of what you have done and for path labs especially know that you are making a difference in someone's life. One condition that makes any job worthwhile is constantly learning as well as constantly teaching others. If you are not and robotics have taken over then you are definitely missing something. Any individual who does not regard themselves as a lifelong student is missing out on a world full of wonders. You need be knowledgeable beyond the level at which you are teaching and you cannot bring students up to your level of knowledge. However given the right attitude and incentives you may be able to direct them to progress beyond your level. Barry From Jodiputnam <@t> aol.com Fri Feb 24 14:56:35 2006 From: Jodiputnam <@t> aol.com (Jodiputnam@aol.com) Date: Fri Feb 24 14:56:46 2006 Subject: [Histonet] CLIA '88 Message-ID: <1e2.4daa5a52.3130cd03@aol.com> Hi. I have a few questions regarding the CLIA certification requirements. I have surfed the net and can't seem to find the info that I need. I need to see what the requirements are (or are going to be) for techs doing gross dissection and dictation. There have been grumblings around work that in 2008 uncertified techs will be unable to do grossing. Also will uncertified techs still be able to do all aspects of histology lab work. I apologize if this topic has been already discussed, but I have been offline for quite some time. Thank you, Jodi From jcox90 <@t> yahoo.com Fri Feb 24 15:10:03 2006 From: jcox90 <@t> yahoo.com (Jill Cox) Date: Fri Feb 24 15:10:13 2006 Subject: [Histonet] Histology Assistant openings in Tempe AZ Message-ID: <20060224211003.39509.qmail@web52105.mail.yahoo.com> Hello Histonetters, Does anyone want to live in beautiful sunny Arizona? I have a few job openings for Histology Assistants, Transcriptionists, Receptionist and Courier. We are opening a private lab in Tempe AZ. Lab will be fully automated with all new state of the art equipment. Great salary and benefits. If interested contact Jill at 602-319-6471. Hope everyone has a wonderful weekend!! From Jodiputnam <@t> aol.com Fri Feb 24 15:41:08 2006 From: Jodiputnam <@t> aol.com (Jodiputnam@aol.com) Date: Fri Feb 24 15:41:19 2006 Subject: [Histonet] Mohs tech Message-ID: <1c3.3ab84e2d.3130d774@aol.com> On a different note (from my CLIA post), I wanted to sent out a notice of a job opening in Nashville Tennessee. I currently have a part time job doing Mohs work for a doctor in Nashville. I have to give this position up due to health reasons. I would have loved to have made this my full time job. It is a brand new lab, all new equipment and a great working environment. The doctor is one of the nicest (and youngest) that I have ever worked for. This is an opportunity to get in on the ground floor of a new practice that is growing rapidly. I work with his partner at my full time job and see the volume of derm specimens growing weekly. I want to stay on long enough to help them get a replacement. This is the first lab that I have worked in (in 15 years of service in 3 states) that is brand spanking new. The staff is small currently but I like that because you get to know one another better and grow. I look forward to going there each week because the atmosphere is so pleasant and you really feel appreciated. Anyway, please email me if you have any questions. Thanks Jodi From Linresearch <@t> aol.com Fri Feb 24 17:33:10 2006 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Feb 24 17:33:25 2006 Subject: Fwd: [Histonet] Perfusion set Message-ID: <255.6f71550.3130f1b6@aol.com> From tpmorken <@t> labvision.com Fri Feb 24 17:47:57 2006 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Feb 24 17:48:16 2006 Subject: [Histonet] Pre-treatment module for immunostainers Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D66C@usca0082k08.labvision.apogent.com> Rene, Just to clarify, the Lab Vision Autostainers (three models) and the Dako Autostainer and Autostainer Plus are all manufactured by Lab Vision. The BioCare Nemesis 7200, Richard Allan 710i, and Axiom are also made by Lab Vision. Each uses the same basic platform and allows fully open reagent use. The PreTreatment Module for deparaffinization and AR allows open reagent use as well. However, some labs opt for "reagent rental" to avoid the captial equipment budget process, so then they are tied into the company for some reagents (whichever company they buy from). Tim Morken Lab Vision - Neomarkers web: www.labvision.com Free webhosting for US State Histotechnology Societies: http://www.labvisioncorp.com/demowebsite/index.cfm From: Rene J Buesa ---------------------------------------------------------------------------- ---- Dear Joyce: Unless you are short in trained personnel and perform more than 100 IHC slides daily, I personally do not see the "advantage" of limiting yourself to a company that will sell you proprietary (and expensive) reagents that will lead you to a "closed system". "Ventana" has a new IHC automatic instrument that also deparafinizes and performs HIER and I have heard both good and bad things about it. I always used Dako and laboratories that I have consulted for also use Dako (up to 3 autostainers) and the "beautity" of the Dako instrument is that it is an "open" technology that you can use even with home made reagents (HIER buffers, Ab diluents) as well as any Ab from any manufacturer. To dewax your slides will take less than 30 minutes (and it can be done with your regular H&E autostainer) and HIER requires, at the most, 1 hour, and you don't have to stand by during that time. I would not commit myself to a single manufacturer product. I hope that this will help you! Ren? J. meint002 wrote: Dear Histos, I am in the process of determining which immunostainer to purchase for our lab. DAKO and Lab Vision are the main vendors at the moment. Lab Vision has a new device that they claim will automate the dewaxing and epitope recovery on your slide before you apply the antibodies. Does anyone have experience with this Pre-treatment module or a device of a similar nature? If so, do they work very well, or are they worth the expense? I think you can use your own reagents for the various buffers - so that could keep the reagent cost down some. Otherwise, they would be happy to sell you their own products. Any advice on this matter would be greatly appreciated. Thank you in advance for your feedback. Joyce Meints Histologist University of Minnesota Paul and Sheila Wellstone Muscular Dystrophy Center MMC 206 420 Washington St. SE Minneapolis, MN 55455-0392 e-mail: meint002@umn.edu lab phone: 612-626-4703 From tkngflght <@t> yahoo.com Fri Feb 24 18:12:00 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Fri Feb 24 18:12:10 2006 Subject: [Histonet] Medical Director job description In-Reply-To: <24435074.1140800256954.JavaMail.root@web21> Message-ID: <20060225001200.16415.qmail@web50914.mail.yahoo.com> http://www.cms.hhs.gov/clia/ http://www.cms.hhs.gov/CLIA/16_Certification_Boards_Clinical_Consultants_&_Laboratory_Directors.asp#TopOfPage Hey Ron- This is at least a place to start...the certifications required for qualification for the Med Dir. job. It's at least a place to start. Cheryl Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one Tech at a time. 281.883.7704 c 281.852.9457 o admin@fullstaff.org Sign up for the FREE monthly newsletter AP News--updates, tricks of the trade and current issues for Anatomic Pathology Clinical Labs. Send a 'subscribe' request to APNews@fullstaff.org. Please include your name and specialty in the body of the email. From gu.lang <@t> gmx.at Sat Feb 25 01:55:10 2006 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Sat Feb 25 01:55:14 2006 Subject: [Histonet] Polioma virus - antibody Message-ID: <000001c639e0$cd341d40$eeeea8c0@SERVER01> Please, can somebody recommend an antibody against Polioma virus, that works on FFPE tissue? We have the Bechmark XT, Ventana, and want to use it on renal biopsies. Thanks in advance Gudrun Lang Linz, Austria From Debbiejsiena <@t> aol.com Sat Feb 25 18:04:37 2006 From: Debbiejsiena <@t> aol.com (Debbiejsiena@aol.com) Date: Sat Feb 25 18:05:26 2006 Subject: [Histonet] bx sponges VS no bx sponges Message-ID: <2c9.420ca5c.31324a95@aol.com> What we have found especially on skin specimens is that the use of sponges produces lots of carryover of solutions. We have found out that the drying artifact that is produced when the first paraffin becomes too saturated with xylene carryover and then the tissues are exposed to mostly xylene at 60C. It is probably a combination of the heat and the xylene. The other times that this can happen is if the solutions on the tissue processor are not consistently full to the fill line on the bottle and this can cause drying out of the tissues in the chamber and lastly if during gross, the tissues that are in cassettes waiting processing are not covered completely with solution. I hope that this helps you out. Debbie Siena, HT(ASCP)QIHC From bridge_870 <@t> yahoo.com Sat Feb 25 23:08:09 2006 From: bridge_870 <@t> yahoo.com (Bridgett Baker) Date: Sat Feb 25 23:08:17 2006 Subject: [Histonet] Histo-tech Message-ID: <20060226050809.24711.qmail@web37012.mail.mud.yahoo.com> Hello histonet, I am a Histology student and really enjoy it!! I am interested to know what kind of degree or education is required and what starting salary can be expected for a Histo-tech? Also, how hard is it to find a job as a Histo-tech, is there much of a demand? Thank you, Bridgett in Texas --------------------------------- Brings words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. From jkiernan <@t> uwo.ca Mon Feb 27 00:26:41 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Mon Feb 27 00:26:48 2006 Subject: [Histonet] removing excess x-gal stain (LONG) References: Message-ID: <44029BA1.131D6E00@uwo.ca> Dear Linda K Bauer, Nobody has replied to you on Histonet for about a week, so a few speculative comments from a non-user of the X-gal technique may be better than nothing at all. This reply has three parts. 1. How can you have "excess nonspecific X-gal stain"? The substrate (X-gal) is hydrolysed by beta-galactosidase and one of the products of hydolysis, "X" (which is short for 5-bromo-4-chloroindoxyl), is rapidly oxidized (by the ferricyanide ions in the incubation medium) to an indigoid dye that rejoices in the name of 5,5'-dibromo-4,4'-dichloroindigo. This particular indigoid dye sticks strongly to the nearest protein molecules and does not diffuse into fat or other lipids, and does not form visible crystals. This is one of the most thoroughly studied reactions in enzyme histochemistry. If the method was carried out correctly there should be no "nonspecific" deposition of 5,5'-dibromo-4,4'-dichloroindigo more than 500nm from a beta-galactosidase enzyme molecule. There are some minor controversies about the sensitivity and specificity of indigogenic methods for lysosomal enzymes (with acidic pH optima), and for many of these enzymes there are alternative histochemical methods. 2. I'm assuming that you are using the indigogenic X-gal method to detect the lac-Z reporter gene, which encodes a bacterial beta-galactosidase that works at pH 7+. This gene is tagged onto genes that encode more interesting proteins in experiments with altered cell lines, transgenic mice etc. Cells that transcribe and translate an added or modified gene also make the bacterial enzyme encoded by the reporter gene. Histochemical detection of a beta-galactosidase active at ph 7+ indicates that the "more interesting proteins" are probably being made in the same cell. The problem with lysosomal enzymes referred to at the end of Part 1 (above) does not arise with indigogenic detection of the lac-Z gene product because the pH of the medium is significantly higher than the pH optima of lysosomal glycosidases. "Normal" (animal) beta-galactosidase activity would be detected only after an unduly prolonged incubation. The false-positive result would also show up in your negative control sections of tissues known not to express the lac-Z gene. If your known-negative controls show no blue-green colour and your transgenics (or others) show "excess nonspecific X-gal stain" there is no "excess". You must be working with a tissue whose cells are putting out lots of the bacterial enzyme. The genetic engineer should thank you for providing evidence that the introduced genes entered the host organism and were successfully transcribed and translated. If you have no known-negative and known-positive control results, ask your boss for more detailed advice. 3. This is an answer to your beautifully concise two-line question, cited below. The indigoid dye is insoluble. It was probably deposited at the site of the enzyme, if the histochemical method was done by the book. John Kiernan Anatomy, UWO London, Canada ---------------------------------------- Linda K Bauer wrote: > > I would like to remove excess nonspecific x-gal stain from whole mount > murine embryos. Does anyone have a suggestion or technique for this? > > Linda(Lin)Bauer > Department of Genetics, Cell Biology, and Anatomy > 985455 Nebraska Medical Center > Omaha, NE 68198-5455 > > Phone: (402) 559-2863 > Fax: (402) 559-4001 > Email: lkbauer@unmc.edu > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dsnider <@t> shrinenet.org Mon Feb 27 07:00:58 2006 From: dsnider <@t> shrinenet.org (Snider, Deanna) Date: Mon Feb 27 07:01:05 2006 Subject: [Histonet] bx sponges VS no bx sponges Message-ID: <84BE46B37B314D409C5A17B7BAB022D666ACFE@IDC-EX-VS01.shriners.cc> I too have found this to be true with bx sponges also. I work with clinically engineered skin, which is much thinner than normal human skin. The specimens had to be kept flat. The sponges created all kinds issues. I ran across a product, called a bx capsule, which is a mesh screen that folds shut(like the bx cassettes). The tissue is held securely, and flat, and the carryover is decreased dramatically and there is no artifact. You can order these from several vendors. Shriners Hospital for Children Research Dept. Cincinnatti, Oh 513-872-6388 CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. From eileen.lonergan <@t> verizon.net Mon Feb 27 07:21:25 2006 From: eileen.lonergan <@t> verizon.net (eileen.lonergan@verizon.net) Date: Mon Feb 27 07:21:28 2006 Subject: [Histonet] Appropriate cassette types Message-ID: <25625309.1141046485781.JavaMail.root@vms064.mailsrvcs.net> Fellow Histonetters, We are a large reference lab that processes various medical facilities tissues. We have a new client that insists on using the small screened biopsy cassettes for every tissue type, including breast, uterus, parotid gland etc. We have ordered the appropriate routine cassettes with the slots for their large tissue but they refuse to use them and the quality of the resulting samples is sub-optimal at best. Our patholgoist has volunteered to speak to them since most of Histology's requests fall on deaf ears. Are there any articles and/or opinions on this issue? I have been in the field for quite a few years and this is the first time I have encountered resistance like this. Most of the grossing is done by a PA who actually does a nice job with the actual grossing, it's just using this screened cassette that we are unable to change. Thanks for any opinions here. Eileen Lonergan HT(ASCP) Eileen Lonergan (207) 749-9617 cell (207) 324-6468 home From sa.drew <@t> hosp.wisc.edu Mon Feb 27 07:30:14 2006 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Mon Feb 27 07:30:20 2006 Subject: [Histonet] Polioma virus - antibody Message-ID: We use the BK/SV40-T antibody from Calbiochem on our FFPE renal biopies and it works well. Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Saturday, February 25, 2006 1:55 AM To: Histonetliste (Histonetliste) Subject: [Histonet] Polioma virus - antibody Please, can somebody recommend an antibody against Polioma virus, that works on FFPE tissue? We have the Bechmark XT, Ventana, and want to use it on renal biopsies. Thanks in advance Gudrun Lang Linz, Austria _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From barbara.bublava <@t> meduniwien.ac.at Mon Feb 27 07:31:01 2006 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Mon Feb 27 07:32:08 2006 Subject: [Histonet] staining collagen References: <1140767886.43febc8e333ac@mail.ugent.be> <006701c6391a$8c899e80$47a9abc1@GERICHTS9XOZZ8> Message-ID: <015d01c63ba2$0ebff640$47a9abc1@GERICHTS9XOZZ8> hi Marc, pikro-aniline blue: add 0,5g Anilinblue or methylenblue to 500ml saturated aquous solution of picric acid. everything else is done like van Gieson Result: collagen, basement membranes and reticulin blue; cytoplasm yellow John Kiernan?s book says reticulin and basement membranes are not stained if you use indigocarmine (0,5 g in 200ml) collagen should be blue-green in this case greetings Barbara ----- Original Message ----- From: "Marc Tjwa" To: "Barbara Bublava" Sent: Sunday, February 26, 2006 4:32 PM Subject: Re: [Histonet] staining collagen Dear Barbara I am also interested in the protocol that you have suggested. Would you please be so kind to send me the protocol by mail? Tnx in advance YS Marc Tjwa >You could use anilineblue instead of acid fuchsin. makes better contrast >and is described in John Kiernans book > >tell me if you can?t find a protocol > >greetings >Barbara, Vienna >----- Original Message ----- >From: "Lobke De Bels" >To: >Sent: Friday, February 24, 2006 8:58 AM >Subject: [Histonet] staining collagen > > > >Von Gieson gives a good contrast for the human eye, but the computer can't >make >a difference between yellow/orange and red. >If I only use acid fushine (or aniline) everything stains pink(blue). > >I have to find a way to stain the connective tissue so the computer can >clearly >make a difference between connective tissue and the other tissue. > >The ideal should be a method that only stains collagen. > > >Many thanks in advance for any help > > > >Lobke De Bels >Department of Morphology >Faculty of Veterinary Medicine >Ghent University >Salisburylaan 133 >9820 Merelbeke >Belgium >lobke.debels@ugent.be > -- ============================================================ Marc Tjwa, MD Center for Transgene Technology and Gene Therapy (CTG) Flanders Interuniversity Institute for Biotechnology (VIB) University of Leuven (KUL) Campus Gasthuisberg, Herestraat 49 B-3000, Leuven, Belgium Tel: +32-16-345775; Fax: +32-16-345990 E-mail: Marc.Tjwa@med.kuleuven.be ============================================================ Disclaimer: http://www.kuleuven.be/cwis/email_disclaimer.htm From ree3 <@t> leicester.ac.uk Mon Feb 27 08:08:26 2006 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Mon Feb 27 08:08:35 2006 Subject: [Histonet] beta-oestrogen receptor Message-ID: Looking for an antibody to beta-oestrogen receptor that works on formalin-fixed paraffin processed tissues of mice, have tried the NOVACASTRA antibody, but no luck to date. Thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER. U.K.... From kmerriam2003 <@t> yahoo.com Mon Feb 27 08:45:30 2006 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Mon Feb 27 08:45:35 2006 Subject: [Histonet] cryosette Message-ID: <20060227144530.24877.qmail@web50313.mail.yahoo.com> Hello all, I received some OCT blocks from an outside vendor and they are embedded in something called a "cryosette". They are really cool. Did a google search for them and nothing came up (how often does that happen?). Anyway - does anyone know who makes these and where I can buy them? Thanks, Kim Kim Merriam Novartis Cambridge, MA --------------------------------- Yahoo! Mail Bring photos to life! New PhotoMail makes sharing a breeze. From abright <@t> brightinstruments.com Mon Feb 27 09:18:39 2006 From: abright <@t> brightinstruments.com (Alan Bright) Date: Mon Feb 27 09:09:41 2006 Subject: [Histonet] cryosette Message-ID: Dear Kim, We manufacture a range, what sizes are you interested in. Best Regards Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: Kim Merriam [mailto:kmerriam2003@yahoo.com] Sent: 27 February 2006 14:46 To: Histonet Subject: [Histonet] cryosette Hello all, I received some OCT blocks from an outside vendor and they are embedded in something called a "cryosette". They are really cool. Did a google search for them and nothing came up (how often does that happen?). Anyway - does anyone know who makes these and where I can buy them? Thanks, Kim Kim Merriam Novartis Cambridge, MA --------------------------------- Yahoo! Mail Bring photos to life! New PhotoMail makes sharing a breeze. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Mon Feb 27 09:59:42 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Mon Feb 27 09:59:43 2006 Subject: [Histonet] staining collagen References: <1140767886.43febc8e333ac@mail.ugent.be> <006701c6391a$8c899e80$47a9abc1@GERICHTS9XOZZ8> <015d01c63ba2$0ebff640$47a9abc1@GERICHTS9XOZZ8> Message-ID: <440321EE.4B966435@uwo.ca> Methyl blue, NOT methylene blue! Methyl blue is one of the components of aniline blue, which is often a mixture of related anionic triphenylmethane dyes. (Methylene blue is a cationic thiazine dye with very different staining capabilities.) The picro-aniline blue methods pick up thin collagen fibres that are missed by van Gieson. The staining pattern is closely similar to picro-sirius red F3B, but aniline blue does not enhance the birefringence of type 1 collagen. John Kiernan London, Canada. ---------------------------------------- Barbara Bublava wrote: > > hi Marc, > > pikro-aniline blue: > > add 0,5g Anilinblue or methylenblue to 500ml saturated aquous solution of > picric acid. > everything else is done like van Gieson > Result: collagen, basement membranes and reticulin blue; cytoplasm yellow > > John Kiernan?s book says reticulin and basement membranes are not stained if > you use indigocarmine (0,5 g in 200ml) collagen should be blue-green in this > case > > greetings > Barbara > > ----- Original Message ----- > From: "Marc Tjwa" > To: "Barbara Bublava" > Sent: Sunday, February 26, 2006 4:32 PM > Subject: Re: [Histonet] staining collagen > > Dear Barbara > > I am also interested in the protocol that you > have suggested. Would you please be so kind to > send me the protocol by mail? > > Tnx in advance > > YS > > Marc Tjwa > > >You could use anilineblue instead of acid fuchsin. makes better contrast > >and is described in John Kiernans book > > > >tell me if you can?t find a protocol > > > >greetings > >Barbara, Vienna > >----- Original Message ----- > >From: "Lobke De Bels" > >To: > >Sent: Friday, February 24, 2006 8:58 AM > >Subject: [Histonet] staining collagen > > > > > > > >Von Gieson gives a good contrast for the human eye, but the computer can't > >make > >a difference between yellow/orange and red. > >If I only use acid fushine (or aniline) everything stains pink(blue). > > > >I have to find a way to stain the connective tissue so the computer can > >clearly > >make a difference between connective tissue and the other tissue. > > > >The ideal should be a method that only stains collagen. > > > > > >Many thanks in advance for any help > > > > > > > >Lobke De Bels > >Department of Morphology > >Faculty of Veterinary Medicine > >Ghent University > >Salisburylaan 133 > >9820 Merelbeke > >Belgium > >lobke.debels@ugent.be > > > > -- > ============================================================ > Marc Tjwa, MD > Center for Transgene Technology and Gene Therapy (CTG) > Flanders Interuniversity Institute for Biotechnology (VIB) > University of Leuven (KUL) > Campus Gasthuisberg, Herestraat 49 > B-3000, Leuven, Belgium > Tel: +32-16-345775; Fax: +32-16-345990 > E-mail: Marc.Tjwa@med.kuleuven.be > ============================================================ From Janet.Bonner <@t> FLHOSP.ORG Mon Feb 27 10:25:02 2006 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Mon Feb 27 10:27:24 2006 Subject: [Histonet] Appropriate cassette types Message-ID: Perhaps if you copy out your letter and send it with the next batch of poorly processed tissue....... ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of eileen.lonergan@verizon.net Sent: Mon 2/27/2006 8:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Appropriate cassette types Fellow Histonetters, We are a large reference lab that processes various medical facilities tissues. We have a new client that insists on using the small screened biopsy cassettes for every tissue type, including breast, uterus, parotid gland etc. We have ordered the appropriate routine cassettes with the slots for their large tissue but they refuse to use them and the quality of the resulting samples is sub-optimal at best. Our patholgoist has volunteered to speak to them since most of Histology's requests fall on deaf ears. Are there any articles and/or opinions on this issue? I have been in the field for quite a few years and this is the first time I have encountered resistance like this. Most of the grossing is done by a PA who actually does a nice job with the actual grossing, it's just using this screened cassette that we are unable to change. Thanks for any opinions here. Eileen Lonergan HT(ASCP) Eileen Lonergan (207) 749-9617 cell (207) 324-6468 home _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eileen_dusek <@t> yahoo.com Mon Feb 27 10:30:48 2006 From: eileen_dusek <@t> yahoo.com (eileen dusek) Date: Mon Feb 27 10:30:51 2006 Subject: [Histonet] H&E staining Message-ID: <20060227163048.60296.qmail@web30010.mail.mud.yahoo.com> Hi Everyone, This is a basic question. How long are tech leaving slides in 95% and 100% ETOH after eosin. I feel we are in too long but would like to get other ideas. Our slides have been very washed out, little to no contrast. I appreciate your help Eileen C. Dusek Edward Hospital --------------------------------- Yahoo! Mail Bring photos to life! New PhotoMail makes sharing a breeze. From kemlo.rogerson <@t> waht.swest.nhs.uk Mon Feb 27 10:38:01 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Mon Feb 27 10:38:13 2006 Subject: [Histonet] Appropriate cassette types Message-ID: Have you asked them why they don't use them? They may have a valid reason, bought a 'job lot', CEO is friends with the Manufacturer? Bet they don't do it just to ruin the tissue and upset you. Ask them (nicely), then deal with the response. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 "One of the benefits of getting older is finding wisdom but so many get of us get older and never find it; sadly that is a waste of a life." This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: Bonner, Janet [mailto:Janet.Bonner@FLHOSP.ORG] Sent: Monday, February 27, 2006 4:25 PM To: eileen.lonergan@verizon.net; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Appropriate cassette types Perhaps if you copy out your letter and send it with the next batch of poorly processed tissue....... ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of eileen.lonergan@verizon.net Sent: Mon 2/27/2006 8:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Appropriate cassette types Fellow Histonetters, We are a large reference lab that processes various medical facilities tissues. We have a new client that insists on using the small screened biopsy cassettes for every tissue type, including breast, uterus, parotid gland etc. We have ordered the appropriate routine cassettes with the slots for their large tissue but they refuse to use them and the quality of the resulting samples is sub-optimal at best. Our patholgoist has volunteered to speak to them since most of Histology's requests fall on deaf ears. Are there any articles and/or opinions on this issue? I have been in the field for quite a few years and this is the first time I have encountered resistance like this. Most of the grossing is done by a PA who actually does a nice job with the actual grossing, it's just using this screened cassette that we are unable to change. Thanks for any opinions here. Eileen Lonergan HT(ASCP) Eileen Lonergan (207) 749-9617 cell (207) 324-6468 home _______________________________________________ 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 From rjbuesa <@t> yahoo.com Mon Feb 27 11:21:16 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Feb 27 11:21:19 2006 Subject: [Histonet] H&E staining In-Reply-To: <20060227163048.60296.qmail@web30010.mail.mud.yahoo.com> Message-ID: <20060227172116.73449.qmail@web61218.mail.yahoo.com> As little time as possible to assure dehydration.Few (3-5) in/out dips in each, except for the last where they could stay for 10-15 secs. This is a "try and err thing". Only make sure that you change the alcohols after 150-200 slides have been stained though. Ren? J. eileen dusek wrote: Hi Everyone, This is a basic question. How long are tech leaving slides in 95% and 100% ETOH after eosin. I feel we are in too long but would like to get other ideas. Our slides have been very washed out, little to no contrast. I appreciate your help Eileen C. Dusek Edward Hospital --------------------------------- Yahoo! Mail Bring photos to life! New PhotoMail makes sharing a breeze. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Mail Bring photos to life! New PhotoMail makes sharing a breeze. From mauger <@t> email.chop.edu Mon Feb 27 11:29:06 2006 From: mauger <@t> email.chop.edu (Joanne Mauger) Date: Mon Feb 27 11:35:55 2006 Subject: [Histonet] Polioma virus - antibody Message-ID: Sally, Where do you get a positive control? Joanne Mauger Children's Hospital of Phila. >>> "Drew Sally A." 02/27/06 8:30 AM >>> We use the BK/SV40-T antibody from Calbiochem on our FFPE renal biopies and it works well. Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Saturday, February 25, 2006 1:55 AM To: Histonetliste (Histonetliste) Subject: [Histonet] Polioma virus - antibody Please, can somebody recommend an antibody against Polioma virus, that works on FFPE tissue? We have the Bechmark XT, Ventana, and want to use it on renal biopsies. Thanks in advance Gudrun Lang Linz, Austria _______________________________________________ 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 From andromeda_tm <@t> libero.it Mon Feb 27 11:40:22 2006 From: andromeda_tm <@t> libero.it (Massimo) Date: Mon Feb 27 11:40:32 2006 Subject: [Histonet] H&E staining References: <20060227163048.60296.qmail@web30010.mail.mud.yahoo.com> Message-ID: <003f01c63bc4$e4ed65f0$20ed1c97@SN300208440005> After Eosin: 10 - 15 seconds in distillate water Quick steps for 50, 75, 95% ETOH (they remove Eosin) About 20 - 30 seconds for 100% ETOH ( for each step of two. You could stop longer in this alcohol than the other ones above) Pay attention!!! That procedure is coming from me: an amateur naturalist. I think it would be better for you to lissen the opinions of the "guru" Histologysts of this ML too!!! Best Regards, Massimo ----- Original Message ----- From: "eileen dusek" To: "histonet" Sent: Monday, February 27, 2006 5:30 PM Subject: [Histonet] H&E staining > Hi Everyone, > This is a basic question. > How long are tech leaving slides in 95% and 100% ETOH after eosin. I feel > we are in too long but would like to get other ideas. > Our slides have been very washed out, little to no contrast. > > I appreciate your help > > Eileen C. Dusek > Edward Hospital > > > --------------------------------- > Yahoo! Mail > Bring photos to life! New PhotoMail makes sharing a breeze. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Malcolm.McCallum <@t> tamut.edu Mon Feb 27 11:46:58 2006 From: Malcolm.McCallum <@t> tamut.edu (Malcolm McCallum) Date: Mon Feb 27 11:47:36 2006 Subject: [Histonet] H&E staining Message-ID: Doesn't Hummason's Histotechnique manual have this in it? Not attacking, just trying to provide a helpful reference! Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Massimo Sent: Mon 2/27/2006 11:40 AM To: eileen dusek Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E staining After Eosin: 10 - 15 seconds in distillate water Quick steps for 50, 75, 95% ETOH (they remove Eosin) About 20 - 30 seconds for 100% ETOH ( for each step of two. You could stop longer in this alcohol than the other ones above) Pay attention!!! That procedure is coming from me: an amateur naturalist. I think it would be better for you to lissen the opinions of the "guru" Histologysts of this ML too!!! Best Regards, Massimo ----- Original Message ----- From: "eileen dusek" To: "histonet" Sent: Monday, February 27, 2006 5:30 PM Subject: [Histonet] H&E staining > Hi Everyone, > This is a basic question. > How long are tech leaving slides in 95% and 100% ETOH after eosin. I feel > we are in too long but would like to get other ideas. > Our slides have been very washed out, little to no contrast. > > I appreciate your help > > Eileen C. Dusek > Edward Hospital > > > --------------------------------- > Yahoo! Mail > Bring photos to life! New PhotoMail makes sharing a breeze. > _______________________________________________ > 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 From jwatson <@t> gnf.org Mon Feb 27 11:55:28 2006 From: jwatson <@t> gnf.org (James Watson) Date: Mon Feb 27 11:55:47 2006 Subject: [Histonet] H&E staining Message-ID: Try looking at the AFIP staining manual "Laboratory methods in Histotechnology" Uses eosin-Phloxine for 2 minutes then two minutes each for all the alcohols and xylenes, very consistent , very easy, with excellent staining. James Watson HT, ASCP Facilities Manager of Histology GNF, Genomics Institute of the Novartis Research Foundation Room C015 858-332-4647 jwatson@gnf.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Malcolm McCallum Sent: Monday, February 27, 2006 9:47 AM To: Massimo; eileen dusek Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E staining Doesn't Hummason's Histotechnique manual have this in it? Not attacking, just trying to provide a helpful reference! Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Massimo Sent: Mon 2/27/2006 11:40 AM To: eileen dusek Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E staining After Eosin: 10 - 15 seconds in distillate water Quick steps for 50, 75, 95% ETOH (they remove Eosin) About 20 - 30 seconds for 100% ETOH ( for each step of two. You could stop longer in this alcohol than the other ones above) Pay attention!!! That procedure is coming from me: an amateur naturalist. I think it would be better for you to lissen the opinions of the "guru" Histologysts of this ML too!!! Best Regards, Massimo ----- Original Message ----- From: "eileen dusek" To: "histonet" Sent: Monday, February 27, 2006 5:30 PM Subject: [Histonet] H&E staining > Hi Everyone, > This is a basic question. > How long are tech leaving slides in 95% and 100% ETOH after eosin. I > feel we are in too long but would like to get other ideas. Our slides > have been very washed out, little to no contrast. > > I appreciate your help > > Eileen C. Dusek > Edward Hospital > > > --------------------------------- > Yahoo! Mail > Bring photos to life! New PhotoMail makes sharing a breeze. > _______________________________________________ > 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 From mparker <@t> epl-inc.com Mon Feb 27 11:55:42 2006 From: mparker <@t> epl-inc.com (Mary Parker) Date: Mon Feb 27 11:55:49 2006 Subject: [Histonet] Uneven Staining Message-ID: Can anyone tell me if they have had uneven staining (H&E) on a linear stainer? This one is relatively new to our lab and we are having a very trying time especially with liver, brain and heart sections. We have done all our usual problem solving and ruled out fixation, improper de-paraffinization, and microtoming artifacts. The pattern is variable, inconsistent and random. When we use the same staining set up and do the slides by hand we get better results. Any suggestions would be appreciated. From eileen_dusek <@t> yahoo.com Mon Feb 27 12:12:27 2006 From: eileen_dusek <@t> yahoo.com (eileen dusek) Date: Mon Feb 27 12:12:31 2006 Subject: [Histonet] thanks for the H&E info Message-ID: <20060227181227.38590.qmail@web30006.mail.mud.yahoo.com> Thank you to everyones response t our H&E problem. I appreciate your inputs. Eileen Dusek --------------------------------- Yahoo! Mail Bring photos to life! New PhotoMail makes sharing a breeze. From PMonfils <@t> Lifespan.org Mon Feb 27 12:45:55 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon Feb 27 12:46:05 2006 Subject: [Histonet] H&E staining Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717671@lsexch.lsmaster.lifespan.org> The responses suggest that some people (most, probably) are using an alcoholic eosin solution and others are using aqueous eosin. We use eosin/phloxine in 80% ethanol, followed by 20 dips in one 95% ethanol and 20 dips in each of three absolute alcohols, then into xylene. "Dips" is not a very precise measurement, but our rate is about 2 dips per second, so the slides are in each dish about 10 seconds. This provides a total of 30 seconds immersion in absolute alcohol with constant agitation, which is plenty to dehydrate a slice of tissue 5 microns thick. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > eileen dusek > Sent: Monday, February 27, 2006 8:30 AM > To: histonet > Subject: [Histonet] H&E staining > > Hi Everyone, > This is a basic question. > How long are tech leaving slides in 95% and 100% ETOH after eosin. I > feel we are in too long but would like to get other ideas. > Our slides have been very washed out, little to no contrast. > > I appreciate your help > > Eileen C. Dusek > Edward Hospital > > > --------------------------------- > Yahoo! Mail > Bring photos to life! New PhotoMail makes sharing a breeze. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From saulsbery.1 <@t> osu.edu Mon Feb 27 12:49:26 2006 From: saulsbery.1 <@t> osu.edu (Anne Saulsbery) Date: Mon Feb 27 12:49:34 2006 Subject: [Histonet] Slide Label Responses Message-ID: <200602271849.k1RInQNu032595@defang19.it.ohio-state.edu> I need some help finding the rubberstamp with letters that can be changed that was bought from Staples. I need to know what the stamp is called, I could only find the date stamp which is too long for the slide. Thanks in advance. Anne From Heather.A.Harper <@t> pcola.med.navy.mil Mon Feb 27 13:30:38 2006 From: Heather.A.Harper <@t> pcola.med.navy.mil (Heather.A.Harper@pcola.med.navy.mil) Date: Mon Feb 27 13:32:02 2006 Subject: [Histonet] Need the CLIAA 88 website link Message-ID: Could somebody send me a link to the CLIAA 88 rules? I just got the updated questions for our CAP list and would like to put a copy in my SOP. I have read the CLIAA 88 rules. Is it just me, but do they need to re-write these rules for histo techs? Sounds so vague and I'm sure on down the road, histo techs will start demanding to be paid more for doing the pathologists job. Thank you in advance. Heather A. Harper Supervisor of Histology Naval Hospital Pensacola, FL 32512 From sandosis <@t> uia.net Mon Feb 27 13:56:31 2006 From: sandosis <@t> uia.net (sandosis@uia.net) Date: Mon Feb 27 13:56:38 2006 Subject: [Histonet] Clearing Colon Fat to Reveal Lymph Nodes Message-ID: <200602271956.k1RJuW933549@smtp4.uia.net> Hello, Could you folks please let me know what methods and reagents you have worked with that do a decent job of overnight clearing the fat from colonectomy specimens to further reveal lymph nodes? Many Thanks, Sandy --------------------------------------------- This message was sent using the UIA Web Mail Server. ULTIMATE Internet Access, Inc http://www.uia.net/ From tp2 <@t> medicine.wisc.edu Mon Feb 27 14:27:28 2006 From: tp2 <@t> medicine.wisc.edu (Thomas Pier) Date: Mon Feb 27 14:28:06 2006 Subject: [Histonet] vWF staining on mouse tissues Message-ID: Hello everyone, I have been told that Dako's rabbit polyclonal vWF will work on mouse tissues. I was wondering if anyone had used it for this in the past. If so, I would appreciate it if you could pass the protocol along. Thanks alot. Tom Pier From Eric.C.Kellar <@t> questdiagnostics.com Mon Feb 27 14:52:38 2006 From: Eric.C.Kellar <@t> questdiagnostics.com (Kellar, Eric C) Date: Mon Feb 27 14:52:55 2006 Subject: [Histonet] RE: Revealing lymph nodes Message-ID: <6843061CE6B98E4B96590D4F299618F801583BAE@qdcws0117.us.qdx.com> Sandy, Hartman's fixative (formerly Davidson's fixative)works about the best for me. It is commercially available from MarketLab and BBC Biochemical to name a few. To prepare it yourself, mix the following - 95% ETOH 1200 ml 37% Formaldehyde 800 ml DH20 1200 ml Glacial Acetic Acid 400 ml The tissue can remain in the fixative for up to 24 hours. Transfer to 10% NBF. The nodes begin to turn opaque-white and reveal themselves after about 30 minutes in this fixative. Eric C. Kellar Histology/Immunohistochemistry Quest Diagnostics Inc. Miami Hello, Could you folks please let me know what methods and reagents you have worked with that do a decent job of overnight clearing the fat from colonectomy specimens to further reveal lymph nodes? Many Thanks, Sandy ============================================================================== The contents of this message, together with any attachments, are intended only for the use of the person(s) to which they are addressed and may contain confidential and/or privileged information. Further, any medical information herein is confidential and protected by law. It is unlawful for unauthorized persons to use, review, copy, disclose, or disseminate confidential medical information. If you are not the intended recipient, immediately advise the sender and delete this message and any attachments. Any distribution, or copying of this message, or any attachment, is prohibited. ============================================================================== From rjbuesa <@t> yahoo.com Mon Feb 27 15:45:29 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Feb 27 15:45:35 2006 Subject: [Histonet] Clearing Colon Fat to Reveal Lymph Nodes In-Reply-To: <200602271956.k1RJuW933549@smtp4.uia.net> Message-ID: <20060227214529.96217.qmail@web61212.mail.yahoo.com> Sandy: I used the following solution to harden the lymph nodes and make them more evident to tact (more "revealed"): a- 100% ethanol -------- 2.0 litres b- distilled water -------- 0.68 litres c- 40% formalin --------- 0.32 litres d- acetic acid ------------ 0.2 litres (Total volume = 3.2 litres) It worked quite well! Hope this will help you! Ren? J. sandosis@uia.net wrote: Hello, Could you folks please let me know what methods and reagents you have worked with that do a decent job of overnight clearing the fat from colonectomy specimens to further reveal lymph nodes? Many Thanks, Sandy --------------------------------------------- This message was sent using the UIA Web Mail Server. ULTIMATE Internet Access, Inc http://www.uia.net/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Mail Bring photos to life! New PhotoMail makes sharing a breeze. From portera <@t> msu.edu Mon Feb 27 15:58:35 2006 From: portera <@t> msu.edu (Amy Porter) Date: Mon Feb 27 15:56:49 2006 Subject: [Histonet] vWF staining on mouse tissues References: Message-ID: <000f01c63be8$f55ea5b0$8e7a0923@HistoJJ> Tom - we used this antibody on mouse, standard Avidin/Biotin Peroxidase protocol using Vector Lab reagents. Pronase E for 10 minutes at 37 prior to staining. Worked really well on mouse tissue. ----- Original Message ----- From: "Thomas Pier" To: Sent: Monday, February 27, 2006 3:27 PM Subject: [Histonet] vWF staining on mouse tissues Hello everyone, I have been told that Dako's rabbit polyclonal vWF will work on mouse tissues. I was wondering if anyone had used it for this in the past. If so, I would appreciate it if you could pass the protocol along. Thanks alot. Tom Pier _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cmalc <@t> unimelb.edu.au Mon Feb 27 17:19:52 2006 From: cmalc <@t> unimelb.edu.au (Cathy Malcontenti-Wilson) Date: Mon Feb 27 17:20:00 2006 Subject: [Histonet] predicament Message-ID: <6.2.1.2.2.20060228100047.027ebcf0@mail.staff.unimelb.edu.au> Dear All Gurus, I have a predicament which I need to overcome. We are doing an immuno for podoplanin on mouse tissues. We do proteinase K for enzymatic retrieval, and Hydrogen peroxide block. The antibody is a syrian hamster anti mouse podoplanin. The secondary antibody is a rabbit anti syrian hamster IgG. We then use the DAKO envision plus kit (for rabbit primaries). In this kit, the tertiary antibody is rabbit ant goat IgG HRP conjugated. Problem: heaps of background staining on the negative and positive. After much trouble shooting, we have found that if we omit only the secondary antibody, we get no backgound. Does this mean that the secondary antibody (rabbit anti syrian hamster IgGs) is sticking non specifically to the tissue and causing the background? We have also tried using normal rabbit serum or FCS as a normal serum block but this makes very little difference. Also some serum and detergent in the antibody diluents makes very little difference. Other controls: if we omit only the primary: we get background staining If we omit all antibodies, we get no staining (so its not the endogenous peroxidases giving the background) If we omit the tertiary antibody only, we get no background staining. If we omit the proteinase K we get background staining. Does anyone have any ideas about how we can reduce this large amount of background staining? Or perhaps we are doing something wrong. Also, does anyone have a good list for the types of controls to do when troubleshooting immuno staining problems? I would love to use this as a resource in our lab. Thanks very much. Cathy From int09018 <@t> alphahunt.com Mon Feb 27 23:37:36 2006 From: int09018 <@t> alphahunt.com (HCS) Date: Mon Feb 27 23:37:51 2006 Subject: [Histonet] looking for microprobe stainer buckets Message-ID: <000601c63c29$17e754c0$6601a8c0@hp> Hi, Does anyone have some extra reagent buckets, Part #15-188-37J, They are the eight containers that sit on top of the fisher microprobe? If you have some extras I would be happy to buy them from you. (I have the black lids so I just need the buckets). LeRoy Brown HT(ASCP)HTL HCS Everson, WA 98247 From int09018 <@t> alphahunt.com Mon Feb 27 23:39:56 2006 From: int09018 <@t> alphahunt.com (HCS) Date: Mon Feb 27 23:41:13 2006 Subject: [Histonet] correction on microprobe buckets Message-ID: <000901c63c29$68b66f30$6601a8c0@hp> OOPS the part number should say 15-188-41. the Reagent buckets. thanks LeRoy Brown From kemlo.rogerson <@t> waht.swest.nhs.uk Tue Feb 28 02:16:46 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Tue Feb 28 02:17:00 2006 Subject: [Histonet] Clearing Colon Fat to Reveal Lymph Nodes Message-ID: Methyl Salicylate; tolerates water clears fat but LN's remain opaque. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 "One of the benefits of getting older is finding wisdom but so many get of us get older and never find it; sadly that is a waste of a life." This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: sandosis@uia.net [mailto:sandosis@uia.net] Sent: Monday, February 27, 2006 7:57 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Clearing Colon Fat to Reveal Lymph Nodes Hello, Could you folks please let me know what methods and reagents you have worked with that do a decent job of overnight clearing the fat from colonectomy specimens to further reveal lymph nodes? Many Thanks, Sandy --------------------------------------------- This message was sent using the UIA Web Mail Server. ULTIMATE Internet Access, Inc http://www.uia.net/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo.rogerson <@t> waht.swest.nhs.uk Tue Feb 28 02:20:14 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Tue Feb 28 02:20:24 2006 Subject: [Histonet] Uneven Staining Message-ID: We had a linear stainer once and uneven staining was a problem; look at dewaxing and complete immersion in haematoxylin and eosin. We ultimately moved to another platform. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 "One of the benefits of getting older is finding wisdom but so many get of us get older and never find it; sadly that is a waste of a life." This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: Mary Parker [mailto:mparker@epl-inc.com] Sent: Monday, February 27, 2006 5:56 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Uneven Staining Can anyone tell me if they have had uneven staining (H&E) on a linear stainer? This one is relatively new to our lab and we are having a very trying time especially with liver, brain and heart sections. We have done all our usual problem solving and ruled out fixation, improper de-paraffinization, and microtoming artifacts. The pattern is variable, inconsistent and random. When we use the same staining set up and do the slides by hand we get better results. Any suggestions would be appreciated. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ree3 <@t> leicester.ac.uk Tue Feb 28 06:41:14 2006 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Tue Feb 28 06:41:21 2006 Subject: [Histonet] RE: beta-oestrogen receptor Message-ID: Looking for an antibody to beta-oestrogen receptor that works on formalin-fixed paraffin processed tissues of mice, have tried the NOVACASTRA antibody, but no luck to date. Thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER. U.K.... From tyler-wellington <@t> northwestern.edu Tue Feb 28 08:29:56 2006 From: tyler-wellington <@t> northwestern.edu (Tyler W.) Date: Tue Feb 28 08:30:00 2006 Subject: [Histonet] staining mouse testis Message-ID: <20060228142956.8813D7E54C@merle.it.northwestern.edu> does anyone have any good recomendations as to how to stain mouse testis. I don't need IHC or anything fancy, just a good general pathology. Any suggestions, wise ones? thank you again, tyler w. From Charles.Embrey <@t> carle.com Tue Feb 28 09:09:41 2006 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Tue Feb 28 09:09:44 2006 Subject: [Histonet] Need the CLIAA 88 website link Message-ID: I don't understand exactly what you mean by "doing the pathologist's job". CLIA '88 deals mostly with management requirements and professional qualifications for "high complexity testing". Histotechs, in general, do not fall under these rules. I can't see why CLIA would re-write the rules. If you can give me some clarification, I may be able to answer you better. Charles Embrey PA(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Monday, February 27, 2006 1:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Need the CLIAA 88 website link Could somebody send me a link to the CLIAA 88 rules? I just got the updated questions for our CAP list and would like to put a copy in my SOP. I have read the CLIAA 88 rules. Is it just me, but do they need to re-write these rules for histo techs? Sounds so vague and I'm sure on down the road, histo techs will start demanding to be paid more for doing the pathologists job. Thank you in advance. Heather A. Harper Supervisor of Histology Naval Hospital Pensacola, FL 32512 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From histo20 <@t> hotmail.com Tue Feb 28 09:17:50 2006 From: histo20 <@t> hotmail.com (Paula Wilder) Date: Tue Feb 28 09:17:57 2006 Subject: [Histonet] Cytology slide batching Message-ID: How does everyone batch their cytology specimens for staining? What similar fluids are run together and in what order? Do you use an automatic stainer or do them manually? Thanks in advance for all responses! Paula Wilder From dennijc <@t> vetmed.auburn.edu Tue Feb 28 09:22:52 2006 From: dennijc <@t> vetmed.auburn.edu (John C. Dennis) Date: Tue Feb 28 09:23:00 2006 Subject: [Histonet] Anti Insect Tubulin Message-ID: <5.1.1.5.1.20060228091841.02fe84f0@mailhost.vetmed.auburn.edu> Dear All Can anyone direct me to a supplier of an antibody against insect tubulin--or any arthropod tubulin. I'd prefer a monoclonal but will use anything that will work in paraffin embedded tissues. Thanks so much, John C. Dennis Auburn University Auburn, Alabama USA From fadiafandi <@t> mac.com Tue Feb 28 09:30:24 2006 From: fadiafandi <@t> mac.com (fadiafandi@mac.com) Date: Tue Feb 28 09:29:40 2006 Subject: [Histonet] Experienced FISH technologists In-Reply-To: <200602271810.k1RIA5O8009165@smtpout.mac.com> References: <200602271810.k1RIA5O8009165@smtpout.mac.com> Message-ID: Potential job opportunities are open for highly experienced, certified technologists (either medical technologists or histotechnologists) who satisfy *all* the following requirements: 1. Over > 3 years of experience performing FISH testing in a high- volume laboratory 2. Ability to recognize and troubleshoot quality control issues related to FISH testing 3. Experience in setting up at least 2 different FISH assays 4. Basic knowledge with screening FISH slides for the common, FDA- approved FISH tests 5. Basic knowledge in operating and maintenance a FISH image analysis system (this is not an absolute requirement) Qualified technologists are invited to reply to this email privately. Our laboratory is located in Miami, Florida, but we will be able to work with you remotely as a consultant. Best regards, Hadi Yaziji, M.D. Ancillary Pathways, LLC. From dsantana <@t> pmaonline.com Tue Feb 28 09:34:50 2006 From: dsantana <@t> pmaonline.com (Santana, Diane) Date: Tue Feb 28 09:35:17 2006 Subject: [Histonet] FNA'S Message-ID: <4C96AA62BADFD81180CB0002A5AD67FB071138A9@MAILPMA> I have recently been asked to do the staining for the radiologist FNA's. It has been a while since I have had to do any cyto prepping. I was wondering what anyone is doing and what fixative, stains they are using? Thanks Diane Santana PMA Haverhill, Mass. From tyler-wellington <@t> northwestern.edu Tue Feb 28 10:13:43 2006 From: tyler-wellington <@t> northwestern.edu (Tyler W.) Date: Tue Feb 28 10:13:49 2006 Subject: [Histonet] staining mouse testis Message-ID: <20060228161343.B5ADD7E54D@merle.it.northwestern.edu> Yeah, thanks everyone for the condescending advice regarding H+E. I took it to be a given, I was merely wondering if anyone had any staining advice that worked particularly well for mouse testis. I apreciate you taking the time to respond and hope you had a good time to take a few cheap jabs at me. Pat yourselves on the back, you're all smarter than I am, clearly. "Tyler W." Sent by: histonet-bounces@lists.utsouthwestern.edu 02/28/2006 08:29 AM Please respond to "Tyler W." To: histonet@lists.utsouthwestern.edu cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] staining mouse testis does anyone have any good recomendations as to how to stain mouse testis. I don't need IHC or anything fancy, just a good general pathology. Any suggestions, wise ones? thank you again, tyler w. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ===========End of original message text=========== From galinadeyneko <@t> yahoo.com Tue Feb 28 10:18:43 2006 From: galinadeyneko <@t> yahoo.com (Galina Deyneko) Date: Tue Feb 28 10:18:50 2006 Subject: [Histonet] Collagen stain Message-ID: <20060228161843.59864.qmail@web33105.mail.mud.yahoo.com> Dear Dr. Kiernan and Barbara, and colleagues, I am also very interested in collagen staining using blue dyes for better contrast for image analysis. For now we use picro-sirius red overnight without counterstaining. Could you send me detailed protocol, please.I mean: time of incubation, wash, differentiation ect.As well could you give me a name of Dr. Kiernan book and where I can buy it. Thank you at advance. Galina Deyneko Novartis Cambridge MA 617-871-7613 histonet-request@lists.utsouthwestern.edu wrote: 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-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@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. Re: removing excess x-gal stain (LONG) (John Kiernan) 2. bx sponges VS no bx sponges (Snider, Deanna) 3. Appropriate cassette types (eileen.lonergan@verizon.net) 4. RE: Polioma virus - antibody (Drew Sally A.) 5. Re: staining collagen (Barbara Bublava) 6. beta-oestrogen receptor (Edwards, R.E.) 7. cryosette (Kim Merriam) 8. RE: cryosette (Alan Bright) 9. Re: staining collagen (John A. Kiernan) 10. RE: Appropriate cassette types (Bonner, Janet) 11. H&E staining (eileen dusek) 12. RE: Appropriate cassette types (Kemlo Rogerson) 13. Re: H&E staining (Rene J Buesa) 14. RE: Polioma virus - antibody (Joanne Mauger) 15. Re: H&E staining (Massimo) 16. RE: H&E staining (Malcolm McCallum) 17. RE: H&E staining (James Watson) 18. Uneven Staining (Mary Parker) ---------------------------------------------------------------------- Message: 1 Date: Mon, 27 Feb 2006 01:26:41 -0500 From: John Kiernan Subject: Re: [Histonet] removing excess x-gal stain (LONG) To: Linda K Bauer Cc: histonet Message-ID: <44029BA1.131D6E00@uwo.ca> Content-Type: text/plain; charset=us-ascii Dear Linda K Bauer, Nobody has replied to you on Histonet for about a week, so a few speculative comments from a non-user of the X-gal technique may be better than nothing at all. This reply has three parts. 1. How can you have "excess nonspecific X-gal stain"? The substrate (X-gal) is hydrolysed by beta-galactosidase and one of the products of hydolysis, "X" (which is short for 5-bromo-4-chloroindoxyl), is rapidly oxidized (by the ferricyanide ions in the incubation medium) to an indigoid dye that rejoices in the name of 5,5'-dibromo-4,4'-dichloroindigo. This particular indigoid dye sticks strongly to the nearest protein molecules and does not diffuse into fat or other lipids, and does not form visible crystals. This is one of the most thoroughly studied reactions in enzyme histochemistry. If the method was carried out correctly there should be no "nonspecific" deposition of 5,5'-dibromo-4,4'-dichloroindigo more than 500nm from a beta-galactosidase enzyme molecule. There are some minor controversies about the sensitivity and specificity of indigogenic methods for lysosomal enzymes (with acidic pH optima), and for many of these enzymes there are alternative histochemical methods. 2. I'm assuming that you are using the indigogenic X-gal method to detect the lac-Z reporter gene, which encodes a bacterial beta-galactosidase that works at pH 7+. This gene is tagged onto genes that encode more interesting proteins in experiments with altered cell lines, transgenic mice etc. Cells that transcribe and translate an added or modified gene also make the bacterial enzyme encoded by the reporter gene. Histochemical detection of a beta-galactosidase active at ph 7+ indicates that the "more interesting proteins" are probably being made in the same cell. The problem with lysosomal enzymes referred to at the end of Part 1 (above) does not arise with indigogenic detection of the lac-Z gene product because the pH of the medium is significantly higher than the pH optima of lysosomal glycosidases. "Normal" (animal) beta-galactosidase activity would be detected only after an unduly prolonged incubation. The false-positive result would also show up in your negative control sections of tissues known not to express the lac-Z gene. If your known-negative controls show no blue-green colour and your transgenics (or others) show "excess nonspecific X-gal stain" there is no "excess". You must be working with a tissue whose cells are putting out lots of the bacterial enzyme. The genetic engineer should thank you for providing evidence that the introduced genes entered the host organism and were successfully transcribed and translated. If you have no known-negative and known-positive control results, ask your boss for more detailed advice. 3. This is an answer to your beautifully concise two-line question, cited below. The indigoid dye is insoluble. It was probably deposited at the site of the enzyme, if the histochemical method was done by the book. John Kiernan Anatomy, UWO London, Canada ---------------------------------------- Linda K Bauer wrote: > > I would like to remove excess nonspecific x-gal stain from whole mount > murine embryos. Does anyone have a suggestion or technique for this? > > Linda(Lin)Bauer > Department of Genetics, Cell Biology, and Anatomy > 985455 Nebraska Medical Center > Omaha, NE 68198-5455 > > Phone: (402) 559-2863 > Fax: (402) 559-4001 > Email: lkbauer@unmc.edu > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 2 Date: Mon, 27 Feb 2006 08:00:58 -0500 From: "Snider, Deanna" Subject: [Histonet] bx sponges VS no bx sponges To: Message-ID: <84BE46B37B314D409C5A17B7BAB022D666ACFE@IDC-EX-VS01.shriners.cc> Content-Type: text/plain; charset="us-ascii" I too have found this to be true with bx sponges also. I work with clinically engineered skin, which is much thinner than normal human skin. The specimens had to be kept flat. The sponges created all kinds issues. I ran across a product, called a bx capsule, which is a mesh screen that folds shut(like the bx cassettes). The tissue is held securely, and flat, and the carryover is decreased dramatically and there is no artifact. You can order these from several vendors. Shriners Hospital for Children Research Dept. Cincinnatti, Oh 513-872-6388 CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. ------------------------------ Message: 3 Date: Mon, 27 Feb 2006 07:21:25 -0600 (CST) From: eileen.lonergan@verizon.net Subject: [Histonet] Appropriate cassette types To: histonet@lists.utsouthwestern.edu Message-ID: <25625309.1141046485781.JavaMail.root@vms064.mailsrvcs.net> Content-Type: text/plain; charset=ISO-8859-1 Fellow Histonetters, We are a large reference lab that processes various medical facilities tissues. We have a new client that insists on using the small screened biopsy cassettes for every tissue type, including breast, uterus, parotid gland etc. We have ordered the appropriate routine cassettes with the slots for their large tissue but they refuse to use them and the quality of the resulting samples is sub-optimal at best. Our patholgoist has volunteered to speak to them since most of Histology's requests fall on deaf ears. Are there any articles and/or opinions on this issue? I have been in the field for quite a few years and this is the first time I have encountered resistance like this. Most of the grossing is done by a PA who actually does a nice job with the actual grossing, it's just using this screened cassette that we are unable to change. Thanks for any opinions here. Eileen Lonergan HT(ASCP) Eileen Lonergan (207) 749-9617 cell (207) 324-6468 home ------------------------------ Message: 4 Date: Mon, 27 Feb 2006 07:30:14 -0600 From: "Drew Sally A." Subject: RE: [Histonet] Polioma virus - antibody To: "Histonet" Message-ID: Content-Type: text/plain; charset="US-ASCII" We use the BK/SV40-T antibody from Calbiochem on our FFPE renal biopies and it works well. Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Saturday, February 25, 2006 1:55 AM To: Histonetliste (Histonetliste) Subject: [Histonet] Polioma virus - antibody Please, can somebody recommend an antibody against Polioma virus, that works on FFPE tissue? We have the Bechmark XT, Ventana, and want to use it on renal biopsies. Thanks in advance Gudrun Lang Linz, Austria _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Mon, 27 Feb 2006 14:31:01 +0100 From: "Barbara Bublava" Subject: Re: [Histonet] staining collagen To: "Marc Tjwa" , Message-ID: <015d01c63ba2$0ebff640$47a9abc1@GERICHTS9XOZZ8> Content-Type: text/plain; charset="iso-8859-1" hi Marc, pikro-aniline blue: add 0,5g Anilinblue or methylenblue to 500ml saturated aquous solution of picric acid. everything else is done like van Gieson Result: collagen, basement membranes and reticulin blue; cytoplasm yellow John Kiernan?s book says reticulin and basement membranes are not stained if you use indigocarmine (0,5 g in 200ml) collagen should be blue-green in this case greetings Barbara ----- Original Message ----- From: "Marc Tjwa" To: "Barbara Bublava" Sent: Sunday, February 26, 2006 4:32 PM Subject: Re: [Histonet] staining collagen Dear Barbara I am also interested in the protocol that you have suggested. Would you please be so kind to send me the protocol by mail? Tnx in advance YS Marc Tjwa >You could use anilineblue instead of acid fuchsin. makes better contrast >and is described in John Kiernans book > >tell me if you can?t find a protocol > >greetings >Barbara, Vienna >----- Original Message ----- >From: "Lobke De Bels" >To: >Sent: Friday, February 24, 2006 8:58 AM >Subject: [Histonet] staining collagen > > > >Von Gieson gives a good contrast for the human eye, but the computer can't >make >a difference between yellow/orange and red. >If I only use acid fushine (or aniline) everything stains pink(blue). > >I have to find a way to stain the connective tissue so the computer can >clearly >make a difference between connective tissue and the other tissue. > >The ideal should be a method that only stains collagen. > > >Many thanks in advance for any help > > > >Lobke De Bels >Department of Morphology >Faculty of Veterinary Medicine >Ghent University >Salisburylaan 133 >9820 Merelbeke >Belgium >lobke.debels@ugent.be > -- ============================================================ Marc Tjwa, MD Center for Transgene Technology and Gene Therapy (CTG) Flanders Interuniversity Institute for Biotechnology (VIB) University of Leuven (KUL) Campus Gasthuisberg, Herestraat 49 B-3000, Leuven, Belgium Tel: +32-16-345775; Fax: +32-16-345990 E-mail: Marc.Tjwa@med.kuleuven.be ============================================================ Disclaimer: http://www.kuleuven.be/cwis/email_disclaimer.htm ------------------------------ Message: 6 Date: Mon, 27 Feb 2006 14:08:26 -0000 From: "Edwards, R.E." Subject: [Histonet] beta-oestrogen receptor To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Looking for an antibody to beta-oestrogen receptor that works on formalin-fixed paraffin processed tissues of mice, have tried the NOVACASTRA antibody, but no luck to date. Thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER. U.K.... ------------------------------ Message: 7 Date: Mon, 27 Feb 2006 06:45:30 -0800 (PST) From: Kim Merriam Subject: [Histonet] cryosette To: Histonet Message-ID: <20060227144530.24877.qmail@web50313.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello all, I received some OCT blocks from an outside vendor and they are embedded in something called a "cryosette". They are really cool. Did a google search for them and nothing came up (how often does that happen?). Anyway - does anyone know who makes these and where I can buy them? Thanks, Kim Kim Merriam Novartis Cambridge, MA --------------------------------- Yahoo! Mail Bring photos to life! New PhotoMail makes sharing a breeze. ------------------------------ Message: 8 Date: Mon, 27 Feb 2006 15:18:39 -0000 From: "Alan Bright" Subject: RE: [Histonet] cryosette To: "Kim Merriam" Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Dear Kim, We manufacture a range, what sizes are you interested in. Best Regards Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: Kim Merriam [mailto:kmerriam2003@yahoo.com] Sent: 27 February 2006 14:46 To: Histonet Subject: [Histonet] cryosette Hello all, I received some OCT blocks from an outside vendor and they are embedded in something called a "cryosette". They are really cool. Did a google search for them and nothing came up (how often does that happen?). Anyway - does anyone know who makes these and where I can buy them? Thanks, Kim Kim Merriam Novartis Cambridge, MA --------------------------------- Yahoo! Mail Bring photos to life! New PhotoMail makes sharing a breeze. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Mon, 27 Feb 2006 10:59:42 -0500 From: "John A. Kiernan" Subject: Re: [Histonet] staining collagen To: Barbara Bublava Cc: histonet@lists.utsouthwestern.edu, Marc Tjwa Message-ID: <440321EE.4B966435@uwo.ca> Content-Type: text/plain; charset=iso-8859-1 Methyl blue, NOT methylene blue! Methyl blue is one of the components of aniline blue, which is often a mixture of related anionic triphenylmethane dyes. (Methylene blue is a cationic thiazine dye with very different staining capabilities.) The picro-aniline blue methods pick up thin collagen fibres that are missed by van Gieson. The staining pattern is closely similar to picro-sirius red F3B, but aniline blue does not enhance the birefringence of type 1 collagen. John Kiernan London, Canada. ---------------------------------------- Barbara Bublava wrote: > > hi Marc, > > pikro-aniline blue: > > add 0,5g Anilinblue or methylenblue to 500ml saturated aquous solution of > picric acid. > everything else is done like van Gieson > Result: collagen, basement membranes and reticulin blue; cytoplasm yellow > > John Kiernan?s book says reticulin and basement membranes are not stained if > you use indigocarmine (0,5 g in 200ml) collagen should be blue-green in this > case > > greetings > Barbara > > ----- Original Message ----- > From: "Marc Tjwa" > To: "Barbara Bublava" > Sent: Sunday, February 26, 2006 4:32 PM > Subject: Re: [Histonet] staining collagen > > Dear Barbara > > I am also interested in the protocol that you > have suggested. Would you please be so kind to > send me the protocol by mail? > > Tnx in advance > > YS > > Marc Tjwa > > >You could use anilineblue instead of acid fuchsin. makes better contrast > >and is described in John Kiernans book > > > >tell me if you can?t find a protocol > > > >greetings > >Barbara, Vienna > >----- Original Message ----- > >From: "Lobke De Bels" > >To: > >Sent: Friday, February 24, 2006 8:58 AM > >Subject: [Histonet] staining collagen > > > > > > > >Von Gieson gives a good contrast for the human eye, but the computer can't > >make > >a difference between yellow/orange and red. > >If I only use acid fushine (or aniline) everything stains pink(blue). > > > >I have to find a way to stain the connective tissue so the computer can > >clearly > >make a difference between connective tissue and the other tissue. > > > >The ideal should be a method that only stains collagen. > > > > > >Many thanks in advance for any help > > > > > > > >Lobke De Bels > >Department of Morphology > >Faculty of Veterinary Medicine > >Ghent University > >Salisburylaan 133 > >9820 Merelbeke > >Belgium > >lobke.debels@ugent.be > > > > -- > ============================================================ > Marc Tjwa, MD > Center for Transgene Technology and Gene Therapy (CTG) > Flanders Interuniversity Institute for Biotechnology (VIB) > University of Leuven (KUL) > Campus Gasthuisberg, Herestraat 49 > B-3000, Leuven, Belgium > Tel: +32-16-345775; Fax: +32-16-345990 > E-mail: Marc.Tjwa@med.kuleuven.be > ============================================================ ------------------------------ Message: 10 Date: Mon, 27 Feb 2006 11:25:02 -0500 From: "Bonner, Janet" Subject: RE: [Histonet] Appropriate cassette types To: eileen.lonergan@verizon.net, histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=iso-8859-1 Perhaps if you copy out your letter and send it with the next batch of poorly processed tissue....... ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of eileen.lonergan@verizon.net Sent: Mon 2/27/2006 8:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Appropriate cassette types Fellow Histonetters, We are a large reference lab that processes various medical facilities tissues. We have a new client that insists on using the small screened biopsy cassettes for every tissue type, including breast, uterus, parotid gland etc. We have ordered the appropriate routine cassettes with the slots for their large tissue but they refuse to use them and the quality of the resulting samples is sub-optimal at best. Our patholgoist has volunteered to speak to them since most of Histology's requests fall on deaf ears. Are there any articles and/or opinions on this issue? I have been in the field for quite a few years and this is the first time I have encountered resistance like this. Most of the grossing is done by a PA who actually does a nice job with the actual grossing, it's just using this screened cassette that we are unable to change. Thanks for any opinions here. Eileen Lonergan HT(ASCP) Eileen Lonergan (207) 749-9617 cell (207) 324-6468 home _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Mon, 27 Feb 2006 08:30:48 -0800 (PST) === message truncated === --------------------------------- Yahoo! Mail Bring photos to life! New PhotoMail makes sharing a breeze. From galinadeyneko <@t> yahoo.com Tue Feb 28 10:49:12 2006 From: galinadeyneko <@t> yahoo.com (Galina Deyneko) Date: Tue Feb 28 10:49:19 2006 Subject: [Histonet] collagen staining Message-ID: <20060228164912.16508.qmail@web33102.mail.mud.yahoo.com> Dear Dr. Kiernan and Barbara, and colleagues, I am also very interested in collagen staining using blue dyes for better contrast for image analysis. For now we use picro-sirius red overnight without counterstaining on mouse and rabbit hearts and arteries.. Could you send me detailed protocol, please.I mean: time of incubation, wash, differentiation ect. As well could you give me a name of Dr. Kiernan book and where I can buy it. Thank you at advance. Galina Deyneko Novartis Cambridge MA 617-871-7613 --------------------------------- Yahoo! Mail Bring photos to life! New PhotoMail makes sharing a breeze. From decorah <@t> rarc.wisc.edu Tue Feb 28 12:40:29 2006 From: decorah <@t> rarc.wisc.edu (Maureen Decorah) Date: Tue Feb 28 12:40:36 2006 Subject: [Histonet] spinal cords Message-ID: > >I have an ongoing study with sheep using a hyperbaric chamber. I >remove the brain and spinal cords but they haven't decided how they >want to process them yet. They are doing NMR on some so we can't >cut them yet. I am looking for a way to bag them individually and >hold them without deterioration. They are presently in a large tub >(in mass) with periodic formalin changes. > >Any suggestions? > >A >-- > >******************************************************************************* >Dr.Annette Gendron-Fitzpatrick Rm. 380 Enzyme Institute >Dir. Pathology and Laboratory Services 1710 University Ave. >Research Animal Resource Center Madison, WI 53705-4098 >University of Wisconsin (608) >262-1239, gendron@rarc.wisc.edu >******************************************************************************* > -- @@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@ Maureen Decorah (608) 262-0933 1710 University Avenue (608) 265-2698 Fax 385 Enzyme Institute Madison, WI 53726-4087 decorah@rarc.wisc.edu @@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@ From rsrichmond <@t> aol.com Tue Feb 28 12:58:12 2006 From: rsrichmond <@t> aol.com (rsrichmond@aol.com) Date: Tue Feb 28 12:58:30 2006 Subject: [Histonet] Revealing lymph nodes In-Reply-To: <200602281301.7c144048ff1143@rly-yi03.mx.aol.com> References: <200602281301.7c144048ff1143@rly-yi03.mx.aol.com> Message-ID: <8C80AB00ED763D6-116C-E2F6@FWM-M15.sysops.aol.com> I think that a clearing fixative (such as the examples already given; there are also commercial products) is essential for finding the small lymph nodes that are often positive for colon cancer in colon resection specimens. Remember that just one tiny positive lymph node upstages the tumor, and makes chemotherapy mandatory - so there's a real patient care decision involved here. In my travels as a locum tenens pathologist I noted that about half of pathologists were using clearing fixatives, and about half were not. As far as I know there have been no official pronouncements on the subject. Clearing fixatives are less necessary in axillary dissection material for breast cancer - the specimensare smaller and the lymph nodes - and metastases - are usually much bigger. The mesenteries from a colon cancer resection are voluminous and hard to search through. Bob Richmond Gastonia NC From TJJ <@t> Stowers-Institute.org Tue Feb 28 13:05:19 2006 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Tue Feb 28 13:05:42 2006 Subject: [Histonet] PECAM antibody staining in chick Message-ID: Has anybody gotten PECAM/CD31 antibody to successfully stain in chick tissue? If so, please provide the antibody and immunostaining conditions that worked for you. Thanks very much! Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From angela.mcnabola.b <@t> bayer.com Tue Feb 28 13:38:27 2006 From: angela.mcnabola.b <@t> bayer.com (Angela McNabola) Date: Tue Feb 28 13:37:35 2006 Subject: [Histonet] BLNK (B cell Linker protein) Message-ID: Hi all, Is anybody out there doing IHC using this antibody? I've tried a few and have had some success on human tissues, but need to try to get it work on mouse tissues as well. Any suggestions would be greatly appreciated. thanks, Angela Angela McNabola MS, HT(ASCP)SLS, QIHC Scientist Bayer Healthcare 400 Morgan Lane West Haven, CT 06516 203-812-5001 fax# 203-812-5820 angela.mcnabola.b@bayer.com _______________________________________________________________________________________________ The information contained in this e-mail is for the exclusive use of the intended recipient(s) and may be confidential, proprietary, and/or legally privileged. Inadvertent disclosure of this message does not constitute a waiver of any privilege. If you receive this message in error, please do not directly or indirectly use, print, copy, forward, or disclose any part of this message. Please also delete this e-mail and all copies and notify the sender. Thank you. For alternate languages please go to http://bayerdisclaimer.bayerweb.com _______________________________________________________________________________________________ From amylee779 <@t> yahoo.com Tue Feb 28 14:25:01 2006 From: amylee779 <@t> yahoo.com (Amy Lee) Date: Tue Feb 28 14:25:06 2006 Subject: [Histonet] pathologist Message-ID: <20060228202501.56900.qmail@web37414.mail.mud.yahoo.com> Hi, I work in a small lab that there is no pathologist in california bay area. Most of time we work on small animal tissue. We need a good pathologist to review our IHC or special stained slides. Could anybody recommend a good one? It's really appreciated if you can include pricing information. Local pathologist is prefered, but distance will be no problem if you highly recommend s/he. Thanks in advance, Amy --------------------------------- Yahoo! Mail Use Photomail to share photos without annoying attachments. From Lchausse <@t> nmh.org Tue Feb 28 15:11:11 2006 From: Lchausse <@t> nmh.org (Chaussey, Leslie) Date: Tue Feb 28 15:12:46 2006 Subject: [Histonet] Automated Special Stainers Message-ID: I'm doing a study for school on automation in the AP lab and was trying to get a comprehensive listing of automated special stain instruments on the market as well as pros & cons for each. If those of you who use automated special stainers could provide the vendor & a few pros & cons, I'd appreciate it. Thanks. ----------------------------------------- This message and any included attachments are intended only for the addressee. The information contained in this message is confidential and may constitute proprietary or non-public information under international, federal, or state laws. 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. From omnivore98 <@t> yahoo.com Tue Feb 28 15:53:52 2006 From: omnivore98 <@t> yahoo.com (Heather Palmer-Renko) Date: Tue Feb 28 15:53:55 2006 Subject: [Histonet] Polioma virus - antibody Message-ID: <20060228215352.70504.qmail@web31315.mail.mud.yahoo.com> In regards to a known diagnostic antibody specific to the polioma virus I don't believe one exist and I would imagine it would be an ASR if so, in addition I know that there has been some research on polioma and they made up their own poly-clonal cocktail. I am sure you already know this but, your first step would be doing a FISH for renal carcinoma to rule that out-I am sure you do. I am VERY familiar with the Ventana XT platform and know that if you could find a concentrate somewhere it could work-have you tried zymed? I will try to look through some literature tonight see what I can find. Good luck! Heather Renko, B.S., HT(ASCP) --------------------------------- Brings words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. From pruegg <@t> ihctech.net Tue Feb 28 16:16:37 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Tue Feb 28 16:16:36 2006 Subject: [Histonet] pathologist In-Reply-To: <20060228202501.56900.qmail@web37414.mail.mud.yahoo.com> Message-ID: <200602282216.k1SMGTU2000294@chip.viawest.net> Amy, I have my own business in Colorado and offer pathology evaluation at $75 per hour, I consult with a board certified pathologist to provide the report. Services and costs related included as an attachment here. I do a lot of pathology consult for IHC for the companies developing the antibody reagents. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Lee Sent: Tuesday, February 28, 2006 1:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] pathologist Hi, I work in a small lab that there is no pathologist in california bay area. Most of time we work on small animal tissue. We need a good pathologist to review our IHC or special stained slides. Could anybody recommend a good one? It's really appreciated if you can include pricing information. Local pathologist is prefered, but distance will be no problem if you highly recommend s/he. Thanks in advance, Amy --------------------------------- Yahoo! Mail Use Photomail to share photos without annoying attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kimrichey <@t> cableone.net Tue Feb 28 20:14:44 2006 From: kimrichey <@t> cableone.net (kimrichey@cableone.net) Date: Tue Feb 28 19:33:36 2006 Subject: [Histonet] seeking Histology Technician position Message-ID: <1141179284_81905@S2.cableone.net> I will be relocating to the Dallas, TX area in May and am very interested in a Histology Technician position. If anyone knows of available positions, helpful contacts, and/or what certifications I need to acquire, I would greatly appreciate it. Thanks, Kim Richey