Re: [Histonet] bone saw for cutting slabs
Hi Merissa, Exakt technologies makes a wonderful saw designed specifically for exactly what you are trying to do with a hack saw. It is a bit pricy though. Contact Linda Durbin at 405-848-5800 for a quote. Alternatively, you can use a wet saw designed for cutting stained glass. Check out the one by Gryphon: Not too expensive cuts bone well if you take your time. Other vendors like Mar-Med make alternative blades for this saw as well. Good luck, ~Sean McBride Scientific Specialist Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 571-989-BONE (m) 412-268-8275 (fax) smcbr...@andrew.cmu.edu On Aug 19, 2014, at 12:45 PM, M.O. wrote: Histoland! Happy Tuesday! I just wanted to get your feedback on cutting slabs from human femora for histopathological analysis. At them moment we are just using a hack saw to cut 7mm slabs from femora. We notice some marks on the cartilage from sawing, so when we cut the tissue down after decalcification for histological preparation, we cut the thickness down to 4mm and remove the damaged tissue. Would using some sort of bone saw damage the tissue even more or would it be comparable to using a hack saw? Is there a saw that you recommend that is precise and easy to handle that doesn't damage tissue greatly? Thank you so much for your help! Sincerely, Merissa ___ 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] change of email address
Hi Linda, I'm changing jobs, so I need to change my histonet email address. The new email address is: seanmcbride...@me.com. Thanks for all of your help, Sean McBride___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Alizarin Red on undecalcified bone
Hi Mesru, 1. Cryosectioning followed by Alizarin red staining may work. I'm not well-versed in this technique, so I will let other experts in this technique guide you. 2. Alizarin red functions by binding to calcium ions. Acids or chelating agents used in de-mineralization processes may very well cleave the alizarin-calcuim bond, resulting in loss of the differentiating stain. If time, materials and specimens are of a crucial nature, I would not recommend experimenting with this technique. 3. For years, I have worked with pmma embedded mineralized bone tissues with much success. There are a variety of stains to assist you in differentiation of various bone related structures. If you are not well versed with these techniques, there are labs, myself included, which can give you some pointers or contract to do the work for you. Best regards, Sean Sean McBride Senior Scientific Specialist Research Histology Services Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 571-989-BONE (m) 412-268-8275 (o) 412-268-8275 (fax) smcbr...@andrew.cmu.edu Dear Histology Experts, I need your help. I am trying to stain undecalcified bone (adult mouse limb) with Alizarin Red S. I was thinking about two options: 1. Fixed frozen sections with Cryojane tape transfer system then staining the sections with Alizarin Red. 2. Alcohol fixed whole limb (whole mount preparation) stained with Alizarin Red then decalcified and processed for paraffin embedding/sectioning. I am not sure if the dye will survive decalcification and paraffin processing procedures. 3. I am trying to avoid doing plastic embedding unless there is no other option. Any insights are highly appreciated. Regards, Mesru ___ 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] question about refrigeration
Hi Peggy, In my opinion, it depends upon the objective of the study. Cell autolysis begins shortly after death as a result of lack of oxygen supply as well as nutrients. The membranes of the lysosomes break down, and the acid hydrolases begin to degrade the cellular If you're merely looking to do a macro analysis, such as the quantitation of bone, you should be fine. However, if you're hoping to get results at the cellular level, either with traditional stains or IHC which relies on intact protein structure, you should immerse the specimens immediately after retrieval into formaldehyde solution. Good luck. Best regards, Sean Sean McBride AFIRM Project Leader Senior Scientific Specialist Research Histology Services Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 571-989-BONE (m) 412-268-8275 (o) 412-268-8275 (fax) smcbr...@andrew.cmu.edu On Feb 28, 2014, at 9:12 AM, DiCarlo, Margaret wrote: Histonetters, I was wondering how long a bone specimen (portion of pt's tibia in this case) can be in a refrigerator before it will decompose and not produce a good quality section? The case was completed at 3:30 pm, refrigerated and then put into formalin at 8:15 am the next day? I am pushing to put specimen into formalin immediately after surgery. Thank you. Peggy DiCarlo HT (ASCP) Ortho Bone Lab Buffalo General Hospital Buffalo, NY 14203 716-859-1293 The Keeping You Informed section of Kaleida Health`s website features a wealth of information, stories and pictures about our valued workforce and the tremendous momentum our organization is experiencing. Check us out at: www.kaleidahealth.org/kyi CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to 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, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please call Kaleida Healths Technology Assistance Center at (716) 859-. ___ 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] Xylene Resistant Labels
Hi Schaundra, I have used the StainerBondz labels from Brady for quite a while, and am quite pleased with them. Good luck! Sean McBride Senior Scientific Specialist Bone Tissue Engineering Center Research Histology Services Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 571-989-BONE (m) 412-268-8275 (o) 412-268-8275 (fax) smcbr...@andrew.cmu.edu On Feb 19, 2014, at 6:06 PM, Schaundra Walton wrote: Hey Histonetters! We are looking to make some cost effective efficiency changes in some of our outlying labs. We have one small lab currently hand writing slides and then printing paper labels and relabeling slides after staining. Does anyone know of xylene resistant labels that could be printed on a desktop printer or protective covers for paper labels? We'd like to eliminate the double labeling system for efficiency reasons. Any suggestions are welcome. Thanks, Schaundra Walton BS, HTL(ASCP) Histology Supervisor PathGroup Nashville, TN ___ 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] Bone samples long-term storage in 10% formalin or 4% paraformaldehyde
Dear Orla, Post fixation, we have stored our bone specimens in 1x PBS while having them sent out for MicroCT analysis. We have also stored them in 1x PBS at 4°C post fixation when necessary until further processing without having adverse affects on our staining. However, we chose conventional staining over IHC for our results. Best regards, ~Sean McBride Scientific Specialist Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-915-1683 (m) 412-268-8275 (fax) smcbr...@andrew.cmu.edu On Dec 9, 2013, at 8:44 AM, Orla M Gallagher wrote: Thanks to everyone for your comments. I may not have been clear in my question - our researchers don't wish to decalcify these formalin-fixed bones yet, but rather to store them for more than a couple of weeks, in case they need to carry out MicroCT followed by histology later. I'm aware that the formalin or paraformaldehyde will degrade over time, but I just wondered if anyone has a protocol for storage without decalcification? I guess transfer to 70% ethanol is an option but this is also not ideal for longterm storage, and would need to be removed before decal in EDTA. All the best, Orla On 6 December 2013 16:12, Wineman, Terra terra.wine...@novusint.com wrote: I would suggest a different protocol if the tissue will not be processed for a while. I would say a week in 10%NBF and then transfer the bones to an EDTA decal solution. The bones will decal slowly without the affects of the formic acid. I am in research and this is what we do with our bones. Terra Wineman, HTL (ASCP)CM Research Biologist 636-926-7476 phone terra.wine...@novusint.com -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto: histonet-boun...@lists.utsouthwestern.edu] On Behalf Of pru...@ihctech.net Sent: Thursday, December 05, 2013 2:50 PM To: gu.l...@gmx.at; 'Orla M Gallagher' Cc: histonet@lists.utsouthwestern.edu Subject: RE: AW: [Histonet] Bone samples long-term storage in 10% formalin or 4% paraformaldehyde i would think u are correct in advising formic acid decal and then processing into paraffin for the best protection of the trap enzyme, immunoreactivity, etc. A couple of weeks in formalin should be fine. Paraformaldehyde show be the same as formalin. I do know a way to restore the enzyme activity for TRAP that may have been lost so if u need that let me know. - Original Message - Subject: AW: [Histonet] Bone samples long-term storage in 10% formalin or 4% paraformaldehyde From: Gudrun Lang gu.l...@gmx.at Date: 12/5/13 11:42 am To: 'Orla M Gallagher' o.m.gallag...@sheffield.ac.uk Cc: histonet@lists.utsouthwestern.edu Paraformaldehyd is formaldehyd in solid form. Formalin is the aequous solution of formaldehyd. So the main characteristics are the same. Gudrun Lang -Urspruuml;ngliche Nachricht- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von Orla M Gallagher Gesendet: Donnerstag, 05. Dezember 2013 19:31 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Bone samples long-term storage in 10% formalin or 4% paraformaldehyde Dear Histonetters, What is your opinion on storing bone samples long-term (more than a couple of weeks) in 10% formalin? As I was taught, best practice has always been to fix only as long as necessary, depending on the size of the sample, then decalcify and process to wax, and I always stress this to everyone I advise. However, research colleagues sometimes wish to do histology on bone samples that have been stored for months ..or even years! As the formalin pH becomes more acidic, there is formalin pigment and the immunoreactivity and TRAP enzyme activity is diminished or destroyed during long fixation, is there any way of minimising this e.g. has anyone tried regularly replacing the old formalin with fresh buffered formalin, or storing formalin-fixed bones in any other medium? I'm also interested in how best to fix in 4% paraformaldehyde and whether the problems are the same with long-term storage. Thanks for your comments. All the best, Orla -- ** Ms. Orla Gallagher Bone Analysis Laboratory Mellanby Centre for Bone Research Department of Human Metabolism D Floor Medical School University of Sheffield Beech Hill Road Sheffield S10 2RX UK Website: http://mellanbycentre.dept.shef.ac.uk Tel: 0044114-2713337 (office) 0044114-2713174 (lab) E-Mail: o.m.gallag...@sheffield.ac.uk *STOP*: Do you really need to print this e-mail? *BE GREEN:* Keep it on the screen. *Times Higher Education University of the Year* Data protection and confidentiality: The information contained in this message or any appended documents may be privileged and confidential and is intended
Re: [Histonet] Mouse bone blocks/sections, bone marrow smears
Hi Jian, I specialize in orthopedic histology, and I also live in Pittsburgh. I would be glad to discuss the details of your project and see if I can be of any assistance to you. Please contact me at your convenience. Best regards, ~Sean McBride Scientific Specialist Bone Tissue Engineering Center Carnegie Mellon Research Institute 700 Technology Drive Pittsburgh, PA 15219-3124 571-989-BONE (m) 412-268-8275 (fax) seanmcbride...@me.com On Mar 21, 2013, at 7:26 PM, Yu, Jian wrote: Dear all, We like to make paraffin embedded bone of mouse tibia and cut longitudinal sections for HE sections. Does anyone have a easy-to-follow protocol to prepare (fix and decal) the bone before processing? We routinely fix soft tissue (ie. mouse GI tract and liver) and send off to our pathology core for processing and embedding. Any tips on cutting the blocks is helpful too. These sections appear brittle. We are also interested in a practical protocol for making mouse BM smears and staining to determine cellularity changes after radiation. We just started working with the bone. Any advice will be very helpful and greatly appreciated. Thanks, Jian *** Jian Yu, Ph.D. University of Pittsburgh Cancer Institute Email: y...@upmc.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] Undecalcified bone IHC
René Jeffery, I don't see sectioning as that big of a challenge. I often thin section mineralized bone sections ~7 microns with my Leica microtome. Although I have a great interest in developing an IHC technique for mineralized bone, I have never had a spare moment to carry out any developmental work. I am concerned more about the effects of the solvents used in hard tissue histology on the antigens/epitopes of interest. Does anyone use IHC staining on PMMA mineralized bone specimens on a regular basis? Best regards, ~Sean McBride Scientific Specialist Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-915-1683 (m) 412-268-8275 (fax) smcbr...@andrew.cmu.edu On Mar 12, 2012, at 11:06 AM, Rene J Buesa wrote: Undecalcified? How are you going to section it? If you can section it, just use any IHC protocol for regular sections. Good luck! René J. --- On Mon, 3/12/12, Jeffery Howery jeffery.how...@jcl.com wrote: From: Jeffery Howery jeffery.how...@jcl.com Subject: [Histonet] Undecalcified bone IHC To: histonet@lists.utsouthwestern.edu Date: Monday, March 12, 2012, 10:59 AM Does anyone have a protocol for Undecalcified bone for IHC? ___ 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] Need SOP for Sanderson's for Thin Section
Hi folks, I have used Sanderson's stain for my thick, ground bone specimens successfully for years, but have never tried it with thin section specimens. I have a thin section study that I need to stain with Sanderson's, but am unsure of staining times and temperatures for these de-plasticized slides. Does anyone have a thin section SOP that they would be willing to share? Thanks! ~Sean McBride Scientific Specialist Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-915-1683 (m) 412-268-8275 (fax) smcbr...@andrew.cmu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] embedding centers
Double ditto. Sakura TEC ~Sean McBride Scientific Specialist Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-915-1683 (m) 412-268-8275 (fax) smcbr...@andrew.cmu.edu On Sep 13, 2011, at 9:40 AM, Sherwood, Margaret wrote: Ditto. Sakura's Tissue-Tek Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (EDR 214) Massachusetts General Hospital 50 Blossom Street Boston, MA 02114-2696 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-1206 (fax) msherw...@partners.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Weaver, Stephanie Sent: Tuesday, September 13, 2011 9:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] embedding centers Hello histonetters! I know this has been asked before, but there's not much in the recent archives. I'd like to see what everyone thinks is now the best paraffin embedding center. They all seem very similar now, and I don't see any one instrument that looks much different from the others. My primary concern is reliability and long working life, but of course I would also like an instrument that is user friendly, ergonomic and affordable. Please let me know if you have a very good experience with any embedding center or especially if anyone has had a particularly bad experience and let me know any features that you find spectacular or useless. Thanks for the advice! Stephanie Weaver Texas Veterinary Medical Diagnostic Laboratory ___ 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
Re: [Histonet] blades
Dorothy, I put ours in a 15 mL centrifuge tube with a cap sit it on the base of the microtome for the next use, that way, no one gets cut the blade is able to be used to the fullest of it's potential. :-) Best regards, ~Sean McBride Scientific Specialist Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-915-1683 (m) 412-268-8275 (fax) smcbr...@andrew.cmu.edu On Jun 24, 2011, at 4:53 PM, Webb, Dorothy L wrote: Trying to clean up some things hanging out there in our lab and wondering what everyone does with a blade that has been used minimally and tech done for the day with the microtome. Where do you store that blade for use tomorrow or do you toss and not worry about the cost involved? I do not like them sitting on top of the microtome. Any good ideas?? Thanks, as always! 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. HealthPartners R001.0 ___ 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] Retirement
Hey Sally, I can attest to what you are saying. :-) I spent two summers as an undergraduate research scientist at the Inhalation Toxicology Research Institute in Albuquerque, and I had an absolutely wonderful experience. The rugged countryside is quite beautiful, and truly enchanting. I would often ride my motorcycle across the various regions of the state, enjoying all the variety that the countryside had to offer. I have many fond memories of those days, so thanks for sharing :-) ~Sean On Jun 17, 2011, at 2:08 PM, Breeden, Sara wrote: It seems that my reference to RETIREMENT has gotten everyone thinking about it. Heh...heh.. It has been suggested that I reconnoiter in advance of Those of You Who Won't Be Retiring Before I Do (February 29, 2012, if the creek don't rise...). I would be happy to perform that hazardous duty but I need more of those $5.00 donations coming in for whatever it was that I posted last week (I hope my gray cells will rejuvenate when I retire). I won't need the money for travel because I think I'm right where I need to be. Have any of you thought about New Mexico??? Just within the past year, it has occurred to me many times why this is such a good choice for retirement. We do not have hurricanes, we do not have tornados (okay, maybe rarely), we are not prone to earthquakes, the weather is jolly darn good 90% of the time (spring is out - way too windy) and we don't have more than a couple or four inches of snow in the winter. We don't start our furnace/heater until November and it's only in use until maybe early April. The air conditioner was just put to use two weeks ago and we won't need it past the first part of September. Low cost of living, lots of homes (reasonably priced - info upon request) for sale and the number of things one can do in New Mexico are practically endless. We have everything but a beach (and if California keeps shaking, we might have beachfront property - not that I'd wish that on California...). We have skiing, a big lake (fondly called Elephant Butt [Butte]) for water activities, stream, river and lake fishing out the kazoo, mountains to climb, white sands in which to wallow, beautiful sunsets and terrible drivers. Oops - that one slipped out! The margarita (and Bud Light) are the State Drinks (if one is so inclined) and this is the Land of Manana (read it like Spanish). Manana is way much better than I needed that right this very minute and no excuses! Shaded patios, cool evening breezes and gorgeous cool summer mornings (at least until 7:00 a.m.). Besides, I need a replacement beginning March 1, 2012. Brand new lab, tech-designed, bright and LEED, tons of space, a separate storage room for blocks and slides AND a volatile storage room with two acid cabinets and two xylene/alcohol cabinets and a salary (that's another subject, I do work for a State...). Can't have everything, but this is darned close. And I do not work for the Chamber of Commerce! Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services 1101 Camino de Salud NE Albuquerque, NM 87102 505-383-9278 (Histology Lab) ___ 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] Grossing Station Recommendation
Colleagues, We are looking to replace our current pathology grossing hood with a new station, so I am looking for recommendations. Thanks in advance for all of your wonderful advice. Best regards, ~Sean McBride Scientific Specialist Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-915-1683 (m) 412-268-8275 (fax) smcbr...@andrew.cmu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] band saws
Hi Hazel, In my opinion, you have a few options for saws and blades but it mostly depends upon your budget and where along the process you intend to use the saw (grossing, trimming pmma blocks, cutting sections for ground work, etc.) EXAKT corporation makes an excellent band saw that uses diamond blades, and Well corporation makes a very reputable diamond wire saw. Both of these pieces of equipment are rather expensive, but are capable of precision work that less expensive models cannot match. For less precise work, I have found that Gryphon Corporation (Model C-40) and Mar-Med (Item#80) make a wonderful little diamond band saw that is very useful. A variety of diamond and tooth blade designs are available, and the cost is quite reasonable. I would be more than glad to give you a few pointers off line to share with you my knowledge of what works for me. Best regards, ~Sean McBride Scientific Specialist Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-915-1683 (m) 412-268-8275 (fax) smcbr...@andrew.cmu.edu On Mar 7, 2011, at 2:10 PM, Horn, Hazel V wrote: I am looking for a band saw to cut our bone tumors. What do I need to be looking for? Power? Size? Other suggestions? Thanks! Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Autopsy/Histology/Transcription Arkansas Children's Hospital 1 Children's WaySlot 820 Little Rock, AR 72202 phone 501.364.4240 fax501.364.3155 visit us on the web at:www.archildrens.org ** 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
[Histonet] Request for Coverslip Removal SOP
Hi colleagues, I need to remove a glass coverslip from a mounted HE biopsy slide with an implant in order to run some biomaterial surface characterization studies. Does anyone have a SOP that they would be willing to share for removing glass coverslips without damaging the specimen? Thanks in advance for all of the great advice that I always get from the histonet. Best regards, ~Sean McBride Scientific Specialist Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-915-1683 (m) 412-268-8275 (fax) smcbr...@andrew.cmu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Correcting van Gieson over-stain
Hi folks, We have a set of slides that we stained in van Gieson's solution, and unfortunately, they turned out too dark. I tried immersion in 100% ethanol for ~2 minutes, and that helped a little, but they are still too dark. Does anyone have any experience or suggestions as to what solution to use to better differentiate the staining? Thanks in advance for all of your suggestions! ~Sean ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Cassette Marking
Nita, We use HistoTec pens by Newcomer Supply ~Sean On Oct 19, 2010, at 2:22 PM, Nita Searcy wrote: ** High Priority ** If you HAVE to manually mark cassettes - what are you using? Cassette pens ? Pencils ? What is the rest of the world doing? Anything else on the market? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Division Manager, Anatomic Pathology 2401 S. 31st. Street 254-724-2438 Temple, Texas, 76502 nsea...@swmail.sw.org 254-724-2438 Nita Searcy.vcf___ 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] Rabbit Brain Tissue Processing Schedule
Hello everyone, While I am well versed in protocols to process bone from a variety of species, my experience in processing soft tissue is less extensive. Currently, I do have some soft tissue protocols which I inherited from my predecessor, and with which I have generally had success, but I am know that there exists more tissue specific protocols than I am currently using. More specifically, I would like to find a protocol to fix, infiltrate and embed rabbit brains in paraffin. Would anyone have a protocol that they would not mind sharing? Thanks again to all of the folks on histonet who share their vast knowledge with those of us who are still learning. Best regards to all, ~Sean Sean McBride Senior Researcher Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-915-1683 (m) 412-268-9807 (fax) smcbr...@andrew.cmu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] (no subject)
Hello everyone, I have an old Leica RM 2065 microtome that is in need of repair (the clutch stopped working), but according to Leica, the machine is no longer supported by the company. Does anyone have any suggestions for a company or technician who might be able to repair the machine? Thanks in advance, ~Sean ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] decalcification
Hi Dorothy, I'm not sure what size bone specimens with which you are working, but I generally work with femurs calvaria from rabbits, canine other small rodents. I fix my specimens longer in 10% NBF using an automated vacuum infiltration processor. Following fixation, if decal is required, I will decal in buffered formic acid solution and use a calcium reagent set to titrate to the decalcification endpoint. If you would like any specifics, contact me offline. Best regards, ~Sean McBride Senior Researcher Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-915-1683 (m) 412-268-9807 (fax) smcbr...@andrew.cmu.edu --- On Mon, 8/2/10, Webb, Dorothy L dorothy.l.w...@healthpartners.com wrote: From: Webb, Dorothy L dorothy.l.w...@healthpartners.com Subject: [Histonet] decalcification To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu Date: Monday, August 2, 2010, 10:14 AM I would appreciate any feedback on what all are using in your decalcification process. We get a lot of large bones in and the past 2-3 months have noticed a huge problem in our microtomy process with these samples. We have been grossing the bones in and leaving the sample in the cassette in 10% formalin for 24 hours befoere placing in decal for up to 8 hours and still having the inner portion of the sample look underprocessed and crunchy! Any suggestions would be appreciated! Dorothy Webb, HT Regions Hospital Technical Supervisor 651-254-2962 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. HealthPartners R001.0 ___ 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] Request for Owner's Manual: Leica SM2500E
Colleagues, I have a Leica SM2500E sliding sledge microtome for which I am searching for a copy of the owner's manual. If anyone has one that they could share with me, I would be most indebted. Best regards, ~Sean McBride Senior Researcher Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-901-7540 (m) 412-268-8641 (fax) smcbr...@cs.cmu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Request for Staining Advice: Rabbit Calvaria underlying Cerebral Hemishpere
Colleagues, I have been charged with the task of evaluating an hydroxyapatite coated biomaterial implanted in the calvaria of a rabbit model. The calvaria and cerebral hemisphere were excised en bloc, and the objective of the experiment is to assess not only new bone formation and osseointegration but also any pathological effects on the dura mater, arachnoid, pia mater and underlying cerebral hemisphere. I have assessed both brain and bone individually in the past, but never en bloc. The specimens have been embedded in pmma and mounted for thick ground section (~30 35 microns) evaluation. Does anyone have any suggestions for staining techniques that will show both bone brain morphology in an en bloc specimen? Kind regards, ~Sean McBride Senior Researcher Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-901-7540 (m) 412-268-8641 (fax) smcbr...@cs.cmu.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Cassettes and Processing and Fixation ~ Oh My!
Laura, 1. Specimen size dictates to cassette size for our lab. 2. Yes, we use sectioned racks organize our specimens in sequential order in case the marker is accidentally solvated during the processing. 3. Again, specimen size and tissue type dictate to our fixation schedule, but we never formalin fixate a specimen for only 5 minutes. I prefer to error on the side of caution. ~Sean McBride Senior Researcher Bone Tissue Engineering Center Carnegie Mellon Research Institute Suite 4311 700 Technology Drive Pittsburgh, PA 15219-3124 412-268-8275 (o) 412-901-7540 (m) 412-268-8641 (fax) smcbr...@cs.cmu.edu On 3/9/09 10:43 AM, Jones, Laura lpjo...@srhs-pa.org wrote: Hi all. We are having a discussion here about everything in my subject line, but to be more specific: 1. Do you all use different types of cassettes for different sizes of tissues? We have screen cassettes for prostate biopsies, and biopsy cassettes for skins, and regular flow through type for larger-than-they-should-be pieces that have grid marks on them. 2. When you load your cassettes on the processor, do you use the organized basket that spaces them out or the random basket? If you use the random method, how tightly do you feel the cassettes can be packed? Isn't it true that if they are packed too tightly, the fixation of the tissue will be compromised? And, how does everyone use agitation/stirring on the processor? We have always used it, but are wondering how others are doing things. 3. Finally, for a run of combination tissues as described above, what would be your recommended time in formalins? We know all Pathologists are in a hurry for slides, but is a 5 minute station ever acceptable? This is all a result of weeks of discussion about possible changes we could make here, and varying things we have learned over the years... we'd just like to hear what everyone else is doing. For instance: would it be simpler to use biopsy cassettes for everything? Should we consider using the random basket instead of the organized one? How far back could we cut our processing times to expedite things? We'd really appreciate the input of all you experts. Thanks in advance! ___ 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
