[Histonet] Paraffin sections or formalin fixed tissue for EM - looking for reference lab
Hi Carol, We can do that here; I am copying an internal e-mail I got from the pathologist that runs our EM. Hope this helps: Regards, Mike *** Mike, Can you follow up on this and give me some idea of projected number of cases? I would be interested in talking with Carol Wilson if you have her phone number. Thanks. Sheldon Sheldon Bastacky, MD Director, Nephropathology and Electron Microscopy Laboratory University of Pittsburgh Medical Center - Presbyterian University Hospital Department of Pathology (Room A610) 200 Lothrop Street Pittsburgh, PA 15213-2582 Phone: (412) 647-9612 Fax: (412) 647-3399 Pager: (412) 392-7205 E-mail: bastack...@upmc.edumailto:bastack...@upmc.edu https://pathconsult.upmc.com/ ** Michael A. Nalesnik, M.D. Professor of Pathology Division of Transplantation and Hepatic Pathology UPMC Montefiore Rm E738 phone 412-647-7645 fax 412-647-5237 Confidential UPMC Health System information. Any unauthorized or improper disclosure, copying, distribution, or use of the contents of this e-mail and attached document(s) is prohibited. The information contained in this e-mail and attached document(s) is intended only for the personal and confidential use of the recipient(s) named above. If you have received this communication in error, please notify the sender immediately by e-mail and delete the original e-mail and attached document(s). By communicating with UPMC staff through e-mail, you agree to comply with UPMC's e-mail terms of use found at http://www.upmc.comhttp://www.upmc.com/. Should you decide that you do not want to comply with these terms, it is your obligation to reply to those UPMC staff members with whom you are corresponding to indicate you do not agree to comply with these terms and cease further communication with UPMC by e-mail. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Paraffin sections or formalin fixed tissue for EM - looking for reference lab
Hi All, Can anyone recommend a reference lab that can use paraffin sections or formalin fixed tissue for EM? Thanks in advance for suggestions. Carol Carol Wilson, HT(ASCP) Associate Scientist III Team Leader/Histopathology Ricerca Biosciences, LLC ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Paraffin sections mouse brain
Hi everyone: While I have sectioned a lot of different stuff, I have not done mouse brain before and I am having considerable difficulty. The brains are fixed and were embedded by our pathology department on their automated system. When cutting, the paraffin cuts nicely but the brain tissue shreds as if the infiltration was insufficient. I have re-embedded one, just melted and new paraffin then vacuum oven for 2 hours. The results were the same. I don't know if it would help to re-embed going back to EtOH. Also, from my distant past I recall that when we were paraffin sectioning whole human larynges, that the face of the block was soaked by placing a wet gauze pad on it. I know this help but don't remember what the solution was on the pad. Any help would be appreciated. Thanks Michael J. Lyon, PhD Otolaryngology Research Labs SUNY Upstate Medical University 750 East Adams Street Syracuse, NY 13210 Voice: 315-464-7253 Fax: 315-464-5572 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Paraffin sections mouse brain
Michael, Do you know or can you get the program they used to process the brain tissue? Also are they whole or slices of brain (at what thciknees)? Pam Marcum - Original Message - From: Michael J. Lyon, Ph.D. ly...@upstate.edu To: Histonet histonet@lists.utsouthwestern.edu Sent: Thursday, January 17, 2013 10:43:02 AM Subject: [Histonet] Paraffin sections mouse brain Hi everyone: While I have sectioned a lot of different stuff, I have not done mouse brain before and I am having considerable difficulty. The brains are fixed and were embedded by our pathology department on their automated system. When cutting, the paraffin cuts nicely but the brain tissue shreds as if the infiltration was insufficient. I have re-embedded one, just melted and new paraffin then vacuum oven for 2 hours. The results were the same. I don't know if it would help to re-embed going back to EtOH. Also, from my distant past I recall that when we were paraffin sectioning whole human larynges, that the face of the block was soaked by placing a wet gauze pad on it. I know this help but don't remember what the solution was on the pad. Any help would be appreciated. Thanks Michael J. Lyon, PhD Otolaryngology Research Labs SUNY Upstate Medical University 750 East Adams Street Syracuse, NY 13210 Voice: 315-464-7253 Fax: 315-464-5572 ___ 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
AW: [Histonet] Paraffin sections for molecular assays
Hi! For our KRAS-testing we clean the microtome with LTK008, especially surfaces that come in contact with the sections. We use one-way-toothpicks, single packed, to pick the slide from the blade and give it directly into a sterile collection tube. 1-10 sections are collected depending on the dimensions of the tumorarea. In this way we don't have to deal with to-clean or not-to-clean water in the waterbath, brushes, forceps etc. Gudrun ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Paraffin sections for molecular assays
Dear Histonetters, Can you tell me what procedures you use to prevent DNA/RNA contamination of tissue sections when handling multiple samples. Do you just wipe down blade holder and blade (or use a new blade) between samples, or do you use more stringent cleaning? Thomas Crowell Diagnostic Development Novartis Molecular Diagnostics 45 Sidney Street Cambridge, MA 02139 USA Phone+1 617 8717460 Fax +1 N.A. thomas.crow...@novartis.commailto:thomas.crow...@novartis.com www.novartis.comhttp://www.novartis.com/ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Paraffin sections for molecular assays
Hi, You will want to do more than just wiping it down. You should use something specifically designed to clean such contamination like RNAase away (Invitrogen if memory serves). You'll want to wipe down any surfaces that come in contact with the block, sections or your hands when sectioning. Don't forget your forceps probe or whatever you use and wear gloves. Just because I'm obsessive compulsive, I dip the block it's self into the stuff. It is best to float the sections on molecular biology grade water or de-ionized water that has been treated with DEPC. Molecular water is cheaper and don't stink like DEPC! Good luck, Amos On Tue, Oct 4, 2011 at 1:00 PM, histonet-requ...@lists.utsouthwestern.eduwrote: Message: 2 Date: Tue, 4 Oct 2011 16:44:17 + From: Crowell, Thomas thomas.crow...@novartis.com Subject: [Histonet] Paraffin sections for molecular assays To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Message-ID: bc3645844188c945830e826f2ff0977d014b5...@023-ch1mpn1-022.023d.mgd.msft.net Content-Type: text/plain; charset=us-ascii Dear Histonetters, Can you tell me what procedures you use to prevent DNA/RNA contamination of tissue sections when handling multiple samples. Do you just wipe down blade holder and blade (or use a new blade) between samples, or do you use more stringent cleaning? Thomas Crowell ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Paraffin Sections
Hi Histonetters, When I cut the paraffin sections in the microtome, I am not getting the whole sections intact, rather some portion in the middle of the sections are brittle. But the paraffin portion surrounds the tissue is smooth, nice and intact. Thanks for your suggestion and help. Peri pnagap...@cau.edu mailto:pnagap...@cau.edu mailto:pnagap...@cau.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Paraffin Sections
Peri, Sounds like poor infiltration to me. What kind of tissue are we talking about, and what processing program did you use? Kathleen Principal Lab Technician Neurotoxicology Labs Dept of Pharmacology Toxicology Rutgers, the State University of NJ 41 B Gordon Rd Piscataway, NJ 08854 Nagappan, Peri wrote: Hi Histonetters, When I cut the paraffin sections in the microtome, I am not getting the whole sections intact, rather some portion in the middle of the sections are brittle. But the paraffin portion surrounds the tissue is smooth, nice and intact. Thanks for your suggestion and help. Peri pnagap...@cau.edu mailto:pnagap...@cau.edu mailto:pnagap...@cau.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
Re: [Histonet] Paraffin Sections
You running particularly small specimens like derm? You may need to decrease time in alcohol. Adding a little ammonia water to your ice bath and soaking after facing the block may help too. --Original Message-- From: Kathleen Roberts Sender: histonet-boun...@lists.utsouthwestern.edu To: Nagappan, Peri Cc: Histonet Subject: Re: [Histonet] Paraffin Sections Sent: Aug 4, 2009 1:40 PM Peri, Sounds like poor infiltration to me. What kind of tissue are we talking about, and what processing program did you use? Kathleen Principal Lab Technician Neurotoxicology Labs Dept of Pharmacology Toxicology Rutgers, the State University of NJ 41 B Gordon Rd Piscataway, NJ 08854 Nagappan, Peri wrote: Hi Histonetters, When I cut the paraffin sections in the microtome, I am not getting the whole sections intact, rather some portion in the middle of the sections are brittle. But the paraffin portion surrounds the tissue is smooth, nice and intact. Thanks for your suggestion and help. Peri pnagap...@cau.edu mailto:pnagap...@cau.edu mailto:pnagap...@cau.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 Sent from my Verizon Wireless BlackBerry___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] paraffin sections
Hi James, While I know that others with more experience are going to reply and have very good insights to add, in my few years of experience I've stored the paraffin sections at room temp. for up to several months. Sections are stored only after baking them in a 60 C for 1 hour. This baking is done after the cut slides have air dried overnight. I have not had problems with the sections falling off. I know that optimum paraffin section storage is a topic of debate - ambient air vs. nitrogen vs. covered in paraffin; 4 C vs. room temp., as well as the length of storage. While I haven't seen any report of sections falling off due to how they are stored, maybe others on this list have. There may be other issues, like what type of slides or paraffin blend you use. In our lab we use Superfrost Adhesive Slides Platinum Line (Mercedes Medical) and Polyfin (paraffin blended with plasticizers). Also, epitope and tissue type may be a factor. I stain for VWF, Beta-catenin, and cytoskeletal markers such as Myosin Heavy Chain and Tropopin I, all on rodent skeletal muscle and heart tissues, and so far haven't had any problems that I can tell are due to how the sections are stored. Hope this helps in some way! Merced --On Wednesday, October 08, 2008 9:31 AM -0700 James Dooley [EMAIL PROTECTED] wrote: I am a beginning to do paraffin section. I need to know the following as I have heard many things and I have received my best advice here. As of this moment I was storing them at 4 degrees following sections. The problem I am having is many of my section have been coming off when I deparaffinize the tissue. I have been baking the tissue for 60 minutes prior to the deparaffiniztion step. 1. How to store them? 2. How long they can be stored? 3. Under what conditions should they be stored? 4. When melting the paraffin how long should this be done for and at what temperature prior to deparaffinization? Thank you, James ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Research Technician II 354 BRB (Lee Lab) / 140 Farber Hall (mail) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 In order to put yourself in someone else's shoes, you must first take off yours. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
