RE: SPAM-LOW: [Histonet] Re: replacement Turtox Internal Ear, cochlea slides
Wow that is going to be a tuff one, back in the day I tried to cut Petris pyramid from the inter ear and was told it was the densest/hardest bone in the body and it sure seemed to be. Even decalcified we had to embed it in plastic and use permanent tungsten carbide D profile knives to cut any sections. I am not at the university anymore so can't access those slides or I would try to help you out. Regards, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. Ste.215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 www.ihctech.net www.ihcrg.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Atoska Gentry Sent: Thursday, July 01, 2010 10:24 AM To: histo...@pathology.swmed.edu Subject: SPAM-LOW: [Histonet] Re: replacement Turtox Internal Ear, cochlea slides Hello! please see below inquiry from a colleague. Thanks! Atoska Our histology class is looking for replacements for old Turtox (Guinea Pig) Internal Ear, cochlea slides and was wondering if anyone knew a company who supplied these slides or tissue blocks? I don't think it has to be guinea pig but I haven't been able to locate internal ear slides from any species. Any help would be appreciated. Thanks! ~~~ Michelle (Shelly) Aono Histology Technician IV CVM AU Staff Advisory Council Representative Auburn University CVM-APP 212 Greene Hall, Auburn, AL 36832 (334) 844-5594 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: SPAM-LOW: RE: [Histonet] Canine bone sectioning trouble
Oh my, I agree with Liz, your samples are way too thick and your process is way too short. Even for 3-4mm thickness bone I fix for 24-72 hours and then decal in 5% formic acid usually overnight on a platform shaker. I assume RDO decals a lot faster but not that fast, I do not use RDO because I think the formic acid which may be slower better preserves the antigens for IHC. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. Ste.215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 www.ihctech.net www.ihcrg.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Liz Chlipala Sent: Wednesday, June 30, 2010 5:36 PM To: Schneider,Lynda S; histonet@lists.utsouthwestern.edu Subject: SPAM-LOW: RE: [Histonet] Canine bone sectioning trouble Overall your fixation, decalcification and processing cycle is too short. The sample size is really big, why do you need it so thick? I would first of all let the samples fix longer 24 to 48 hours or I would trim them so that they are about 3-5 mm in thickness. Either way I like fixing bone samples for 24 to 48 hours. I'm not that accustomed to using RDO decal it is hydrochloric acid based and is quicker but even bone that size would take a while to decal. We normally use a formic acid based decal and a sample size that large my take up to 2 weeks to decal. Also to process a sample that size I would be at 4 to 6 hours a station. We frequently process porcine and goat knees in 2 x 3 blocks the samples are quite thick about a cm in thickness and we will process them at 4 to 6 hours a station. My recommendation would be to decal the larger cube of bone for a period of time until you can section through it. I would then cut a 3-4 mm thick piece off the cube and process it at 1 hour per station. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Schneider,Lynda S Sent: Tuesday, June 29, 2010 10:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Canine bone sectioning trouble Hello out there... We are having trouble sectioning canine bone. The samples are bone marrow cubes approximately 2cm thick and 1in wide. They were fixed for 15 hours in 10% NBF and decaled for 30 - 45 mins in RDO rapid decalcifier. They were faced in and then surface decaled again for about 30 mins. When sectioned, much of the marrow was missing and the bone was torn and shredded. We thought that maybe the samples had not been fixed sufficiently so refixed overnight in 10% NBF again. The samples were then reprocessed as they had been originally. The processing schedule used was: 70% EtOH 30mins, 80% EtOH 30mins, 95% EtOH x 2, 30mins each, 100% EtOH x 2, 30 mins each, xylene x 2, 45 mins each, paraffin x 4, 40 mins each. Again they were faced in and decaled for another 30 mins or so. This time as soon as the sections are placed on the water bath (38 degrees) they explode and/or come apart so severely sections can almost not be picked up. If sections are even obtainable they are of horrible quality. Does anyone have any suggestions? Thank you so much in advance! Lynda ___ 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
RE: SPAM-LOW: Re: [Histonet] Chilling Tray for Blocks
Yep that is what I do, I use the plastic containers I get salad or other stuff in I would throw away, fill it and freeze it, I add some water to it and put the blocks on the frozen tray and they get cold and hydrated at the same time. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. Ste.215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 www.ihctech.net www.ihcrg.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kim Merriam Sent: Thursday, July 01, 2010 6:38 AM To: Sherwood, Margaret; Jay Lundgren; Laurie Colbert Cc: histonet@lists.utsouthwestern.edu Subject: SPAM-LOW: Re: [Histonet] Chilling Tray for Blocks we use something like this: https://www.vwrsp.com/catalog/product/index.cgi?catalog_number=414004-256in E=1highlight=414004-256 we keep several sizes in the freezer, depending on how many blocks we need to cut. The nice thing is that it is cheap and you can hydrate your blocks at the same time you are chilling them. Kim Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA From: Sherwood, Margaret msherw...@partners.org To: Jay Lundgren jaylundg...@gmail.com; Laurie Colbert laurie.colb...@huntingtonhospital.com Cc: histonet@lists.utsouthwestern.edu Sent: Wed, June 30, 2010 4:35:53 PM Subject: RE: [Histonet] Chilling Tray for Blocks We actually use a styrofoam box (used for shipping) and it stays cold longer with very little melting. We keep a couple in the -20 freezer. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jay Lundgren Sent: Wednesday, June 30, 2010 4:01 PM To: Laurie Colbert Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Chilling Tray for Blocks Fill a Tupperware container with water, pop it in the freezer, and tomorrow you will have a portable block chiller, with the blocks at a perfect level for your needs, for 1/100th the price. Not trying to be a smarta$$, but Jay A. Lundgren, M.S., HTL (ASCP) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information in this e-mail is intended only for the person to whom it is addressed. If you believe this e-mail was sent to you in error and the e-mail contains patient information, please contact the Partners Compliance HelpLine at http://www.partners.org/complianceline . If the e-mail was sent to you in error but does not contain patient information, please contact the sender and properly dispose of the e-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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: SPAM-LOW: Re: [Histonet] Unstained Slides
I do not bake my unstained slides for IHC controls and store them at 4dc. I bake them just before use. They do last longer this way but Richard is right, there is no telling because it all depends on the fixation, target protein stability and storage conditions. It is always a good idea to cut fresh sections if the stored unstained sections do not perform as expected. Regards, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. Ste.215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 www.ihctech.net www.ihcrg.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Richard Cartun Sent: Monday, June 28, 2010 10:55 AM To: histonet@lists.utsouthwestern.edu; rick.garnh...@memorialhealthsystem.com Subject: SPAM-LOW: Re: [Histonet] Unstained Slides We bake our unstains at 60 degrees C. for 30 minutes prior to filing at RT. There is no set rule for stability. It all depends on fixation, the nature of the protein target, and storage conditions. I've seen absolutely spectacular immunoreactivity on unstained slides stored at RT for 20 years (and longer) and I've seen reduced immunoreactivity in unstained slides stored for as little as 4 weeks. We will always attempt to stain unstained slides when available; however, if the lesion or tumor is negative, and there is no internal control, you better cut fresh sections. Richard Richard W. Cartun, MS, PhD Director, Histology Immunopathology Director, Biospecimen Collection Programs Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 Office (860) 545-2204 Fax rick.garnh...@memorialhealthsystem.com 6/28/2010 11:54 AM Histoland, How is everyone storing/filing unstained slide. And how long are they good for to use for immunohistochemistry. Rick Garnhart HT(ASCP) Memorial Health System Histology Supervisor 1400 E. Boulder St. Colorado Springs, CO 80909 Cell: 719-365-8357 Ph: 719-365-6926 Fax: 719-365-6373 rick.garnh...@memorialhealthsystem.com Mission: To provide the highest quality health care Vision: To create an outstanding health system where patients heal and people thrive Values: Compassion - Integrity - Quality - Respect - Teamwork www.memorialhealthsystem.com The information contained in or attached to this electronic message is privileged and confidential, intended only for the use of the individual(s) named above. If the reader of this message is not the intended recipient, or the employee or agent responsible to deliver it 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 inform the sender immediately and remove any record of this 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
