Re: [Histonet] Plastic coverslip Loosening
Thank you for your help. This is encouraging. Helen -Original Message- From: Glover, Kimberly [mailto:kimberly.glo...@polysciences.com] Sent: Tuesday, March 22, 2016 11:38 AM To: Helen Fedor <hfe...@jhmi.edu> Cc: histonet <histonet@lists.utsouthwestern.edu> Subject: RE: Plastic coverslip Loosening Helen, Submerge the slides with coverslip loosely attached in fresh xylene to reactivate the film resin. Allow to sit in xylene for approximately 1 minute. Afterwards remove slide and press down on a paper towel. This action will allow the film to reattach to the slide while pressing out any excess xylene from the slide. As for the film (containing tissue) that has completed detached from the slide, place the film against a clean slide and place in a slide basket. Submerge slide in fresh xylene and proceed with same procedure above. Sometimes the issue of loosening or lifting of film is seen over time when the coverslipped slides are placed in temperature/humidity conditions that are contrary to the manufacturer's recommendations. Check packaging for storage conditions recommended. I hope this helps. Kimberly A. Glover Life Sciences Product Manager Polysciences, Inc. 400 Valley Road Warrington, PA 18976 www.polysciences.com -Original Message- From: Helen Fedor [mailto:hfe...@jhmi.edu] Sent: Tuesday, March 22, 2016 11:06 AM To: Glover, Kimberly Subject: RE: Plastic coverslip Loosening The film. -Original Message- From: Glover, Kimberly [mailto:kimberly.glo...@polysciences.com] Sent: Tuesday, March 22, 2016 10:25 AM To: Helen Fedor <hfe...@jhmi.edu> Subject: RE: Plastic coverslip Loosening Are you using plastic coverslipping film or the plastic coverslips? Kimberly -Original Message- From: Helen Fedor via Histonet [mailto:histonet@lists.utsouthwestern.edu] Sent: Tuesday, March 22, 2016 9:45 AM To: HISTONET (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Plastic coverslip Loosening Hello, I did try and search the archives to see if there was any advice on this but didn't find an answer. Do you know of a good way to fix H slides that had plastic coverslips where the coverslip is lifting from the slide and in some cases coming completely off? In all cases the tissue is on the coverslip. Thanks in advance. Helen L. Fedor ___ 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] Plastic coverslip Loosening
Hello, I did try and search the archives to see if there was any advice on this but didn't find an answer. Do you know of a good way to fix H slides that had plastic coverslips where the coverslip is lifting from the slide and in some cases coming completely off? In all cases the tissue is on the coverslip. Thanks in advance. Helen L. Fedor ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] MTA-1 Microarrayer
Hello Bernice, we have tried out several and found that enercell works. CR2450 their number is 23-808. WE might have found them on amazon. Cannot remember. Helen L. Fedor Oncology Tissue Services, Manager Johns Hopkins University 411 N. Caroline St Room 310 Basement| Bond St Annex Building Baltimore, MD | 21231 410-614-1660 http://tmalab.jhmi.edu/ -Original Message- From: Bernice Frederick [mailto:b-freder...@northwestern.edu] Sent: Monday, May 04, 2015 10:53 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] MTA-1 Microarrayer Hello all, Anybody have any idea where to get the batteries for this arrayer? Regular 2450 batteries are too thick. Batteries plus was a no go. See some on Amazon but not sure if they are thin enough. Thanks, Bernice Bernice Frederick HTL (ASCP) Senior Research Tech Pathology Core Facility Robert. H. Lurie Cancer Center Northwestern University 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 b-freder...@northwestern.edumailto:b-freder...@northwestern.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] TMA Arrayers
Hello, We have the Pathology Devices Semi automated units. We are happy with them. Our decision to keep with the less automated units are largely due to our work flow. We make TMA's for customers. But the customer has to do all of the work in the design and data entry into out TMAJ Database. I think all of the automated units have data input features that are not compatible with this work flow. Good luck in choosing. Helen L. Fedor Oncology Tissue Services, Manager Johns Hopkins University 411 N. Caroline St Room 310 Basement| Bond St Annex Building Baltimore, MD | 21231 410-614-1660 http://tmalab.jhmi.edu/ -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sally Ann Drew Sent: Thursday, April 30, 2015 1:36 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] TMA Arrayers I was wondering how many people have semi-automated or automated tissue microarray equipment? I am especially interested in those who graduated from manual methods to the automated and how they reviewed and ranked the limited options out there. Thank you! _Sally Sally Ann Drew, MT(ASCP) UWSMPH-Dept. of Pathology TRIP Lab Manager Translational Science BioCore CSC, L5/181 608.265.1093 ___ 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] Vacuum processing of biopsies.
Hello all, I would be interested in pros and cons of using one of the Rapid vacuum processing units for processing of biopsy tissues. Is this technology improving the processing? Any specific problems that may be occurring. Thanks in advance. Helen L. Fedor Oncology Tissue Services, Manager Johns Hopkins University 411 N. Caroline St Room 310 Basement| Bond St Annex Building Baltimore, MD | 21231 410-614-1660 http://tmalab.jhmi.edu/ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Cryosectioning undecalcified bone
Hello, I believe that the products for the CryoJane tape transfer are still available from Fisher. Helen L. Fedor Oncology Tissue Services, Manager Johns Hopkins University 411 N. Caroline St Room 310 Basement| Bond St Annex Building Baltimore, MD | 21231 410-614-1660 http://tmalab.jhmi.edu/ -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: Thursday, February 05, 2015 1:08 PM To: Orla M Gallagher; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Cryosectioning undecalcified bone Orla There is an article in the Journal of Histotechnology from a while ago from John Tarpley that addressed methods for this. I have a pdf of it and I will send in another e-mail. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell l...@premierlab.com www.premierlab.com March 10, 2014 is Histotechnology Professionals Day Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Orla M Gallagher Sent: Thursday, February 05, 2015 10:56 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cryosectioning undecalcified bone Dear Histonetters, We would like to develop cryosectioning for undecalcified mouse and rat bones. Previous attempts to use the CryoJane system from Instrumedics with an old Bright cryostat and solid tungsten carbide blade a few years ago didn't result in successful reproducible sections - marrow without bone or bone without marrow on the slide, in spite of various freezing and prep. protocols used by my colleague. I suspect the old cryostat and blade weren't helping either. Have you any recommendations on the best cryostat to use to do this? We'd like to also use the cryostat for standard soft tissue sectioning. I've seen Leica mentioned in a few papers relating to CryoJane. There appears also to be a tape transfer method from Kawamoto's Section-Lab Co. Ltd. (i...@section-lab.jp) Does anyone use this method or know whether the consumables are still available for purchase, as the website seems dormant? Thanks for any advice, 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. http://www.shef.ac.uk/faculty/medicine-dentistry-health/2.9274/emailwording http://www.sheffield.ac.uk/humanmetabolism/greenimpact http://www.shef.ac.uk/faculty/medicine-dentistry-health/2.9274/emailwording http://www.sheffield.ac.uk/visitors/mapsandtravel ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Cassette Labeler
Hello, we recently purchased the Primera unit from Creative waste solutions. http://www.cwsincorp.com/ we like it. I would be happy to discuss it further if you like. Helen L. Fedor Oncology Tissue Services, Manager Johns Hopkins University 411 N. Caroline St Room 310 Basement| Bond St Annex Building Baltimore, MD | 21231 410-614-1660 http://tmalab.jhmi.edu/ -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Debbie Granato Sent: Monday, January 26, 2015 12:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cassette Labeler Our lab is currently looking into purchasing a cassette and slide labeler. We are a small lab and are looking for a stand- alone unit with the flexibility to be used with the computer or bar code scanner at a later date. I would appreciate any feedback or suggestions for any model that may fulfill our needs. Thank you, Debbie Granato HT(ASCP) ___ 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] Re: Histonet Digest, Vol 130, Issue 16
We had the PTL, which is a little more cumbersome. We are now using the BBP33, and are very happy with it. Helen L. Fedor Prostate Tissue Bank, Manager Oncology Tissue Services, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 410-614-1660 (Marburg) 443-287-7338 (Bond St.) http://tmalab.jhmi.edu/ http://prostatebiorepository.org/ -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Dorothy Hu Sent: Thursday, September 18, 2014 3:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Histonet Digest, Vol 130, Issue 16 Hi Dear Histoneters, Can you tell me which kind of Brady labeler you are using? I want to get one. Do you have catalog number? Many thanks. Dorothy MGH Endocrine histocore I use the Brady system and I like it a lot. I work in a small animal diagnostic lab and will have 60-80 blocks on most days. It saves me time and even better I don't have to read my writing in the morning. I use it for the cassettes and slides mainly but I label reagents I mix with it to. I find it very versatile. Roberta Horner HT/HTL Animal Diagnostic Lab Penn State University Subject: [Histonet] Slide and cassette printers? What are all of you using these days? What would be good for a small histo lab that has the potential to grow? (I am already aware of what Leica and Thermo have from the archives here, but I am still interested in your opinions, of course.) Has anyone tried the printers, labels and attachment machine from Brady? Thank you all so much, Kathleen Principal Lab Technician Histopathology Lab Office of Translational Sciences Rutgers, the State University of NJ 41 B Gordon Road Piscataway, NJ 08854 (848) 445-1443 FAX (732) 445-6905 ___ 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] Slide and cassette printers?
We are also using the Brady labeler. It is versatile, easy to use, they have great tech support. We are not using the cassette label attaching machine. For the cassette printer we are using the Primera. The Tech support there is also great. Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Roberta Horner Sent: Wednesday, September 17, 2014 8:20 AM To: Kathleen Roberts; histonet Subject: RE: [Histonet] Slide and cassette printers? I use the Brady system and I like it a lot. I work in a small animal diagnostic lab and will have 60-80 blocks on most days. It saves me time and even better I don't have to read my writing in the morning. I use it for the cassettes and slides mainly but I label reagents I mix with it to. I find it very versatile. Roberta Horner HT/HTL Animal Diagnostic Lab Penn State University -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Kathleen Roberts Sent: Tuesday, September 16, 2014 5:05 PM To: histonet Subject: [Histonet] Slide and cassette printers? What are all of you using these days? What would be good for a small histo lab that has the potential to grow? (I am already aware of what Leica and Thermo have from the archives here, but I am still interested in your opinions, of course.) Has anyone tried the printers, labels and attachment machine from Brady? Thank you all so much, Kathleen Principal Lab Technician Histopathology Lab Office of Translational Sciences Rutgers, the State University of NJ 41 B Gordon Road Piscataway, NJ 08854 (848) 445-1443 FAX (732) 445-6905 ___ 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] RE: rolling sections
Hi, I think that it is not necessary to actually get them to roll. We just collect all of the sections and put them into the tube. Scrunched, not rolled. Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Roberta Horner Sent: Thursday, June 12, 2014 10:42 AM To: Histonet (histonet@lists.utsouthwestern.edu) Subject: [Histonet] rolling sections I have some researchers that want to do PCR. They want 10 - 10u sections in a micro-centrifuge tube. The only way to get the sections in the tube is for the sections to roll. How do you get sections to roll when you want them to roll? I've tried room temperature, on ice, brand new sharp blade, dull blade and I can still get some really nice ribbons. When I want a thick ribbon it will roll, darn that Murphy and his laws. Roberta Horner Animal Diagnostic Lab Penn State University ___ 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] Temporary position
Hello, We are in search of a temporary histo-tech or histologist to work in our OTS Core lab. Primarily we do research work to support the efforts of the Cancer Center Investigators. A majority of the work is on animal tissues. Gross, Process, Embed, section and do recuts, i.e., lining up blocks. The lab also makes TMA's and coring paraffin blocks for RNA and DNA extractions. Please contact me if you are interested, or know someone that may be interested in this opportunity. Thanks, Helen 410.614.1660 http://tmalab.jhmi.edu/ http://prostatebiorepository.org/ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] embedding cells in parrafin #2
Making cell tissue blocks Cells: . Harvest and fix in formalin* for at least 3hrs or overnight (10 fold more formalin) . HAVE A NICE VISIBLE SIZE PELLET (1 confluent T75 minimum or 1 T175) . Remove formalin* and wash 2x's in cold 1x PBS . Cover cells with some 1x PBS and refrigerate Materials for blocks: 1. Toothpick or 20ul pipette tip 2. Agarose (2% solution agarose with 1xPBS) ultra pure agarose cat#15510-027 from core 3. Cells in 1x PBS 4. 0.5ml microtubes (sterile) 5. Tissue cassettes (1 or 6 compartiment cassettes -Electron Microscopy Science 800-523-5874 cat.# 70078-W) 6. New razor blades 7. Clean cutting board 8. Container to submerge cassette into with enough 1x PBS to cover cassettes Method: . Suspend cells in 1xPBS - use just enough to pipette cells (1:1 dilution is MAX) . Make 2% agarose solution - keep in water bath (42C) so doesn't solidify . Make 200ul agarose plug in 0.5ml tube and wait 3 minutes to solidify . Add *120ul of cell solution to .5ml tube and then add *150ul of agarose and pipette up/down ~3 times to mixed well - CUT END OF PIPETTE TIP OFF JUST A LITTLE SO YOU DON'T DESTROY CELL IF NECESSARY . Add toothpick and let sit 5 minutes . Remove plug with toothpick and cut into sections (if necessary) with new blade on clean surface and add to cassette . If you have extra cell plugs then store then in 1x PBS in frig. * Varies according to how much cell suspension you have. ALWAYS add at least 30ul more of agarose in order for it to make a nice plug. IMPORTANT NOTES: . ALWAYS be sure to label your cassette with an experiment number because that's what the slides will be labeled with. . Be SURE to write on BOTH the top of the cassette and down the sides what cell type is in each compartment (if using a 6 compartment cassette) . Tell the histologist what you fixed your cells in - if fixed in formalin, ethanol, paraformaldehyde, etc. - this is vital for the processing/embedding process . ALWAYS put control cell types in the block - +/- . ALWAYS prepare your block in an asymmetric pattern THIS IS A MUST!! or you will not be able to identify which cell type is placed where on the slide Helen L. Fedor Prostate Tissue Bank, Manager Oncology Tissue Services, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 410.614.1660 http://tmalab.jhmi.edu/ http://prostatebiorepository.org/ -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Mike Tighe Sent: Monday, December 02, 2013 12:06 PM To: histonet@lists.utsouthwestern.edu (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Embedding cells in Paraffin Has anyone tried to embed cells grown in tissue culture? I am trying to put some tissue culture cells through same stress as tissue would go through. Fixation, dehydration, and heat. Any ideas? I could re-suspend in OCT and then fix for extended time with NBF but that doesn't quite seem fair. Thanks for any ideas! ___ 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] The Good Old Days...
You can tell it is Friday. :) Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cristi Rigazio Sent: Friday, September 20, 2013 1:40 PM To: Davis, Cassie Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] The Good Old Days... Hear hear! I agree and was just saying I love hearing the stories (although I am not young). Thanks all for sharing these memories and lessons! Sent from my iPhone On Sep 20, 2013, at 10:13 AM, Davis, Cassie cda...@che-east.org wrote: I enjoy hearing sincere reminiscing...Even though us kids don't know how good we have it, some of us enjoy having an old tech beside us on the bench. I find weeding through the sarcasm can be profitable and in doing so have learned so much. What the old techs did on a daily basis, we only did in the practice lab and when the automated instruments and pre-made solutions that we have come to rely on fail, experience is so very valuable. Only by their blood, sweat and tears have we benefitted however, we have so far to go, let's do it together. Cassandra Davis cda...@che-east.org 302-575-8095 Confidentiality Notice: This e-mail, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please delete this message, and reply to the sender regarding the error in a separate 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Re: Two Patient Identifiers on slides
Hello, I may not have read every e-mail on this discussion but would like to chime in here. I was just having this discussion with someone yesterday. At large research hospitals that have various goals in our mission statements, we are concerned about continuously improving the process to minimize any errors in patient care. But at the same time we also are using these same cases for research. Our IRB's allow us to use the Pathology Archives as a bank of tissues and slides. If the names of patients were on the slides and or blocks that would really make it impossible to do Specimen based research. We really couldn't use any of those materials for research. It has been a process here at Hopkins to try and keep both of these things moving forward, Improving patient care and maintaining access to specimens for research. Helen L. Fedor Prostate Tissue Bank, Manager Oncology Tissue Services, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 410.614.1660 http://tmalab.jhmi.edu/ http://prostatebiorepository.org/ -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bob Richmond Sent: Wednesday, July 31, 2013 7:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Two Patient Identifiers on slides Realizing we're talking about regulator compliance rather than good sense here - nonetheless let a pathologist observe that having the patient's name on the slide is a very important defense against mixing up cases at sign-out. Small labs still usually have no identifier on the slide other than a hand-scribbled accession number (two of the three small labs I'm working in at the moment don't - both CAP accredited), and it's easy for the pathologist to misread the label. I saw a mixup like this just a few days ago - it wasn't my case, but I should have caught the misreading when I reviewed the case, which involved a squamous carcinoma that wasn't. Good management, bad medicine. Bob Richmond Samurai Pathologist Maryville 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] Block labeling
Hello, we are using the labels from the Brady label maker with good success. They are extra sticky and you can find some that will stick to paraffin. We use ones that qualify for freezing since they need to be safe on the ice tray. http://www.bradycorp.com/ Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Patrick Laurie Sent: Monday, June 24, 2013 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Block labeling Hi Everyone, What is the best way to label blocks that are already embedded? We have a large control bank that we are trying to relabel and make search able, however, most marking pens don't write well on paraffin covered blocks, even when we scrape them off. Pencils work, not always the most legibly, so I was wondering if there is a clever solution that someone might have found. We've tried putting stickers or labels on the back, but that falls right off. Thanks in advance for any suggestions. -- Patrick Laurie(HT)ASCP QIHC Histology Manager Celligent Diagnostics, LLC 101 East W.T. Harris Blvd | Suite 1212 | Charlotte, NC 28262 Work: 704-970-3300 Cell: 704-266-0869 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Control Storage
Hello, We are storing our unstained slides at -20 in Ziploc bags. Our TMA blocks are being stored at 4 degrees. Helen L. Fedor Oncology Tissue Services, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 410.614.1660 http://tmalab.jhmi.edu/ http://prostatebiorepository.org/ -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: Wednesday, June 12, 2013 11:56 AM To: Herring, Colleen; Histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Control Storage We store our blocks at room temp, our IHC slides are stored in the fridge. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308 (303) 682-3949 office (303) 881-0763 cell (303) 682-9060 fax l...@premierlab.com Ship to address: Premier Laboratory, LLC 1567 Skyway Drive, Unit E Longmont, CO 80504 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Herring, Colleen [colleen_herr...@bshsi.org] Sent: Wednesday, June 12, 2013 9:48 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Control Storage Does anyone have any ideas or suggestions about the storage of blocks and precut slides for immunos? 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Out of my comfort zone...
We have been using store bought gallons of distilled water in our water baths. This water has been boiled so enzyme activity should be absent. Helen 410.614.1660 http://tmalab.jhmi.edu/ http://prostatebiorepository.org/ Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sue Hunter Sent: Wednesday, April 03, 2013 10:55 AM To: Sarah Dysart; histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Out of my comfort zone... Hello Just wanted to add one more thing - we actually use a dedicated pyrex dish (maybe 6x10 inches) for our water bath for RNA sections. We use warm tap water, but you can put it in the microwave for a short bit if it needs to be warmer. You can spray the dish with RNAse away and wipe before filling with water. Sue -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sarah Dysart Sent: Tuesday, April 02, 2013 5:36 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Out of my comfort zone... So...I have been asked to do some micro-dissection on some slides and then do downstream RT/PCR on them. My molecular knowledge doesn't go much out of the world of IHC so...here is my question... Has anyone ever substituted Citrate Buffer pH6 (or whatever HIER solution you are using) for proteinase K for use in RNA isolation and then later PCR? Does this work? The main question is will the HIER step take off the formalin linkage from the nucleic acids, or just the protein? One last thing is what else goes into these solutions other than Citrate Buffer and Tween? I haven't made it up in forever, I have just been ordering it from companies...I know...lazy... Thanks Sarah Goebel-Dysart, BA, HT(ASCP), QIHC (ASCP) Histotechnologist Mirna Therapeutics 2150 Woodward Street Suite 100 Austin, Texas 78744 (512)901-0900 ext. 6912 ___ 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: [Histonet] Validation of a new tissue processor
You will also need to validate all of the IHC stains and special stains that you do on these tissues. You can take samples from each tissue type and run them in parallel on both machines. Then test all of your special stains and antibodies that you have in your inventory. :) Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Wednesday, November 28, 2012 4:47 PM To: Howe, Helena; 'histonet@lists.utsouthwestern.edu' Subject: Re: [Histonet] Validation of a new tissue processor Using at least 25 different types of tissues, prepare parallel almost identical pairs of slices for each tissue and process them simultaneously in your new tissue processor and in the old one. Prepare HE sections and give them to the pathologists concealing the used tissue processor. The pathologists should decide if there are differences between the pairs of sections. The pathologist's opinion should be registered in the QC file and that will be your validation. René J. From: Howe, Helena helena.h...@promedica.org To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu Sent: Wednesday, November 28, 2012 2:57 PM Subject: [Histonet] Validation of a new tissue processor Does anyone know what the CAP is requiring to validate a new tissue processor? Thank You Helena Howe ProMedica Laboratories _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ EMAIL CONFIDENTIALITY NOTICE This Email message, and any attachments, may contain confidential patient health information that is legally protected. This information is intended only for the use of the individual or entity named above. The authorized recipient of this information is prohibited from disclosing this information to any other party unless required to do so by law or regulation and is required to destroy the information after its stated need has been fulfilled. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution, or action taken in reliance on the contents of this message is strictly prohibited. If you have received this information in error, please notify the sender immediately by replying to this message and delete the message from your system. ___ 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: [Histonet] Need better results from older archival slides?
Agreed! -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Monday, November 19, 2012 11:16 AM To: Rob Day; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Need better results from older archival slides? Rob: If you are really seeking for a confirming study, publish your data and probably there will be a confirmation study. I went to the website you indicated and it seems to me that your original posting is just a subtle and delicate advertisement. Just my opinion otherwise, why don't you tell us all what is that you do to obtain your rejuvenating results? René J. From: Rob Day r...@foliobio.com To: histonet@lists.utsouthwestern.edu Sent: Monday, November 19, 2012 11:03 AM Subject: [Histonet] Need better results from older archival slides? Hello, Having problems getting good Immuno staining and in-situ hybridization results with older slides? I am writing to see if histonet can suggest any scientists who may be interested in helping us with a study related to a slide rejuvenation process we are developing. We think we have found a way to significantly improve the antigenicity of proteins in older archival FFPE preserved slides. Initial tests seem to indicate that our process can allow slides of 5-10 years age, possibly older, to respond to IHC analysis as if they were cut that day. The process may have additional uses and benefits too, including better preservation of cut sections, enhanced ISH results, stabilization of labile proteins and standardization / optimization of IHC staining protocols. We are excited about this development, but we are seeking some independent labs to help us verify our findings. Please feel free to call me to discuss, or pass on this email to any investigators you may know of who are exploring ways to make better use of older archival slides. Folio Biosciences is a provider of biosamples and a commercial research lab based in Columbus Ohio. See: http://www.foliobio.com/. Regards, Rob Day. Rob Day Business Development Folio Biosciences 1476 Manning Pkwy, Powell, Ohio 43065 Direct Line: (614) 407-4547 | Main Office Phone: (614) 846-2809 | Fax: (877) 591-1815 http://foliobio.com www.linkedin.com/in/robdaybiotech This message, including any attachments, is confidential and may be privileged or may contain health information protected by state and federal law. Information and opinions expressed in this message and/or attachments are those of the author and are not necessarily those of the company. If you are not the intended recipient, please notify the sender and delete this message from your system. Any use of this information by individuals other than the intended recipient is strictly prohibited. ___ 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] Bliss Imaging system
Hello, We have a Bliss imaging system that we are no longer using. I has a great Zeiss axiophot microscope with high quality lenses. I believe that Olympus is now the company that has taken over the Bacus Labs Imaging system. Please contact me if you have a desire to acquire this system from us or know of anyone who might be interested in it.. Thanks for your consideration, Helen L. Fedor Prostate Tissue Bank, Manager TMA Core Lab, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 410.614.1660 http://tmalab.jhmi.edu/ http://prostatebiorepository.org/ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Beecher
http://www.pathologydevices.com/TMArrayer.htm You can contact Ron Gebing at Pathology Devices. Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bernice Frederick Sent: Wednesday, July 11, 2012 12:14 PM To: Fellow HistoNetters Subject: [Histonet] Beecher All ,I know this came up before,but who took over Beecher? We need to order some more punchers. Thanks, Bernice Bernice Frederick HTL (ASCP) Senior Research Tech Pathology Core Facility ECOGPCO-RL Robert. H. Lurie Cancer Center Northwestern University 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 b-freder...@northwestern.edumailto:b-freder...@northwestern.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] Billing 88342
That is so true.! -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Paula Pierce Sent: Tuesday, July 03, 2012 1:36 PM To: Jay Lundgren; Victor A. Tobias Cc: HISTONET Subject: Re: [Histonet] Billing 88342 Where is the like button? Thumbs up! Paula K. Pierce, HTL(ASCP)HT President Excalibur Pathology, Inc. 8901 S. Santa Fe, Suite G Oklahoma City, OK 73139 405-759-3953 Lab 405-759-7513 Fax www.excaliburpathology.com From: Jay Lundgren jaylundg...@gmail.com To: Victor A. Tobias vtob...@uw.edu Cc: HISTONET histonet@lists.utsouthwestern.edu Sent: Tuesday, July 3, 2012 12:05 PM Subject: Re: [Histonet] Billing 88342 I am neither a lawyer nor a health care administrator, but, in my experience, the Pathologist picks the (hopefully) most diagnostic blocks from the multiblock cases and submits them for IHC. If you do the requested IHC on, say, 4 blocks out of 30, you charge x4 for the technical fee. After all, you are using 4 times the supplies (buffer, antibody, etc.). Before you hit the cash paying patient with a bill, their primary care provider should warn them what it's going to cost. I have seen a good Pathologist only select one block for IHC when the clinician previously informed him that the patient had no insurance and was paying out of pocket. I think it's interesting that people control *their own* health care costs when no insurance company or the government is involved. Sincerely, Jay A. Lundgren, M.S., HTL (ASCP) On Tue, Jul 3, 2012 at 11:43 AM, Victor A. Tobias vtob...@uw.edu wrote: Looking for other opinions from those who do consult/referral work. If a client sends in a request for a single antibody done on multiple blocks on a single specimen, do you bill the client for each tech component ? The client will do the interpretation. What happens in the above scenario if the request is to bill the patient? Knowing you get reimbursed for one, do you eat the other charges are make the client select the one block? We have run numbers on potential lost revenue and the number is significant. Victor Victor Tobias HT(ASCP) Clinical Applications Analyst Harborview Medical Center Dept of Pathology Room NJB244 Seattle, WA 98104 vtob...@u.washington.edumailto:vtob...@u.washington.edu 206-744-2735 206-744-8240 Fax = Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. ___ 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
RE: [Histonet] TMA blocks
Hello Colleen, We use standard size molds (2.5cm X 3.5cm) that are a litte deeper (7mm) in our core to make the recipient blocks. 2.5cm X 3.5cm. and our TMA with the 1.0mm punch have 13 rows and 16 columns = 208 cores. You can contact me if you want more information. Helen L. Fedor Tissue Microarray Lab, Manager Prostate Biorepository, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 410.614.1660 http://tmalab.jhmi.edu/ -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Colleen Forster Sent: Monday, February 20, 2012 5:02 PM To: Histonet Subject: [Histonet] TMA blocks Hey Histonetters! We have the first generation of the Beecher machine, manual version. We would like to do a super TMA that would hold 180+1.0mm punches. When you do a single TMA the tissue area is 1x1 and out largest mold is 3x3. Has anyone ever trimmed the TMA's down and embedded 2 or 3 of them into one big block? Thanks in advance!! Colleen Forster Bionet Histology Laboratory U of MN, AHC ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Casual Temp Position
Hello, We have a position available for a casual temp employee. This is a reference lab that primarily focuses on animal specimens. We will consider someone who has 2 years or more cutting experience. Please forward your resume to me. thanks Helen 410.614.1660 http://tmalab.jhmi.edu/ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Casual Temp Position
Hello, We have a position available for a casual temp employee. This is a reference lab located in Baltimore MD, that primarily focuses on animal specimens. We will consider someone who has 2 years or more cutting experience. Please forward your resume to me. thanks Helen 410.614.1660 http://tmalab.jhmi.edu/ Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Helen Fedor Sent: Monday, January 09, 2012 1:37 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Casual Temp Position Hello, We have a position available for a casual temp employee. This is a reference lab that primarily focuses on animal specimens. We will consider someone who has 2 years or more cutting experience. Please forward your resume to me. thanks Helen 410.614.1660 http://tmalab.jhmi.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
[Histonet] RE: to cut or not to cut mouse brains
Hello Terry, These are very difficult. The colder the better. (-27). keep the slides in the cryostat. Cut the section. Place section on a cold slide, and keep the slide in the croystat during the following procedure. Hold slide in left hand and brush in right. Place finger under one edge of the section and it will start to thaw, anchoring it to the slide. Gently brush out folds as you continue to advance the thaw line with your fingers under the slide. It takes a bit of practice, and not really perfect. But it works. Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of McLaughlin, Terry Sent: Thursday, November 17, 2011 2:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] to cut or not to cut mouse brains Hello All, I am having trouble cutting frozen mouse brains and was wondering if someone can offer some help. The mouse was perfused in 4% PFA , the brain removed and kept in PFA for 22 hrs. We placed it in 30% sucrose for 20 hrs, until the brain sank in this solution. It was frozen back by using a culture plate floating in liquid nitrogen with OCT . The problem I am having is many folds, and air bubbles under the tissue. What I have done so far: Cut at temp of -25, then tried - 22 and -17. Cut at 10um, then tried 16-25um. Tried making the knife colder by adding dry ice to it for a few seconds. Added dry ice to sample for a couple seconds. The sample cuts fine, it appears to happen when picking up on a slide. I tried picking up by starting at one side or end and letting tissue spread onto slide. Did not work. Also tried gently laying slide on top of sample and removing gently-still folds and air bubbles. Any help would be greatly appreciated as we have 6 more to do. Signed, Help! Terry Kokas ___ 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] to cut or not to cut mouse brains
When the blocks are not croyprotected you can cut them at warmer temperatures. But when they are cryoprotected with the 30% sucrose, they are soft at -18, that is why you need the colder temperature. they will compress at the warmer temperatures and give you a lot of wrinkles. Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Grantham, Andrea L - (algranth) Sent: Thursday, November 17, 2011 2:57 PM To: McLaughlin, Terry Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] to cut or not to cut mouse brains I'm interested in this answer because this has happened to me - but not always. I usually cut brains at warmer temps. like -17 or -18 as opposed to -25 and they are usually handled in the same manner before being brought to the lab frozen in OCT. Andi On Nov 17, 2011, at 12:24 PM, McLaughlin, Terry wrote: Hello All, I am having trouble cutting frozen mouse brains and was wondering if someone can offer some help. The mouse was perfused in 4% PFA , the brain removed and kept in PFA for 22 hrs. We placed it in 30% sucrose for 20 hrs, until the brain sank in this solution. It was frozen back by using a culture plate floating in liquid nitrogen with OCT . The problem I am having is many folds, and air bubbles under the tissue. What I have done so far: Cut at temp of -25, then tried - 22 and -17. Cut at 10um, then tried 16-25um. Tried making the knife colder by adding dry ice to it for a few seconds. Added dry ice to sample for a couple seconds. The sample cuts fine, it appears to happen when picking up on a slide. I tried picking up by starting at one side or end and letting tissue spread onto slide. Did not work. Also tried gently laying slide on top of sample and removing gently-still folds and air bubbles. Any help would be greatly appreciated as we have 6 more to do. Signed, Help! Terry Kokas ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: need help with TMA
Hello, Do 30 minutes at 37 degrees on a glass slide. take from oven and apply gentle pressure to center of block and slide complex, cool to RT and repeat 1 or two times. Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Patricia F Lott Sent: Friday, October 28, 2011 1:13 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] need help with TMA We have recently started doing tissue microarrays. The problem we have is that some of the small cores drop out after the first few sections. I think my problem is in the step where we are supposed to melt the donor and recipient blocks together. We tried 5 minutes at 37 degrees, and then minutes at 60 degrees x3, with cooling to room temp in between. Any suggestions? Thanks, Patty Lott, UAB ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Paraffin sections for molecular assays
Hello, For DNA, we use a new blande and we wipe down microtome, handles, front of blade holder with 95% ethanol on a gauze sponge. For RNA, use a new blade and wipe down all with RNAzap including the water bath and rinse with DEPC water. Use clean water in bath, distilled will work or DEPC water. Depending on the stringency of the assay. You can sometimes get away with a little less. But some of the assays that are being run can cost up to $1,000.00. So the time spent cleaning up is justified. Make sure you are wearing gloves for both RNA and DNA, during the whole process, even when handling the slides .(including the process of labeling slides). I usually lay the slides out on pape rtowels on the lab bench while labeling them. Helen Fedor Johns Hopkins University TMA and Prostate Spore Lab Manager JHU SOM Uro/Pathology 600 N. Wolfe Street, Marburg Room 406 Baltimore MD, 21287-7065 410-614-1660 / Fax 410-614-3695 Pager 410-283-3419 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] on behalf of Crowell, Thomas [thomas.crow...@novartis.com] Sent: Tuesday, October 04, 2011 12:44 PM To: histonet@lists.utsouthwestern.edu Subject: [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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Control Slides
Hello, We have been storing our slides in small zip loc bags at -20 and have
data suggesting that slides stored in this fashion are reasonably well
preserved after 3- 5 years. This was done with TMA slides. Was not done for
special stain slides but for IHC.
Helen
-Original Message-
From: histonet-boun...@lists.utsouthwestern.edu
[mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of
pru...@ihctech.net
Sent: Tuesday, March 15, 2011 8:56 AM
To: Harrison,Sandra C.
Cc: histonet@lists.utsouthwestern.edu; Amador, Amanda
Subject: RE: [Histonet] Control Slides
I do not melt dry my cut control slides until i need to use them,=d
i do store them in boxes at 4c, i have recently run into this problem
=th blocks, so now i seal all my blocks with paraffin and store
them in th�ridge as well.
Patsy
Original Message
Subject: RE:=istonet] Control Slides
From: Harrison, Sandra C. [1]sandra.harris...@va.gov
Date: Mon,=rch 14, 2011 1:40 pm
To: Lee Peggy Wenk [2]lpw...@sbcglobal.net, Amador, Amanda
l=[3]aama...@ameripath.com,=;[4]histo...@lists.utso uthwestern.edu
We dry the control slide at the same time tha=e dry the tissue
being
stained, since controls are supposed to be han=ed in the same
manner as
the test tissue. That being said, I guess if = were truly following
that rule, we would cut the control on the same �y we cut the test
sample. :-)
Other people say that, after they dry the slide, they dip the slide
in paraffin, to cover the
tissu=
so
that the air doesn't touch the tissue during the months before=e
slide
is
used in a stain.
-Original Message-
From: [5]histone=boun...@lists.utsouthwestern.edu
[[6]mailto:histonet-bounces@lists.utsouthwestern.e=] On Behalf Of
Lee
Peggy Wenk
Sent: Thursday, March 10, 21 6:39 PM
To: Amador, Amanda; [7]histonet@lists.utsouthwestern.edu
Subject: Re: [Histone= Control Slides
I've noticed that our spirochete control slides do=t stain as
intense
starting about 3 months after they are sectioned, =d that they stop
staining by about 6 months. (This is with a silver s=in.)
I've talked with other histotechs, and they say they've seen =e
same
phenomenon with AFB and Gram controls. (We use ours up too qui=ly -
we
never have 6-12 months old control slides for these, so I can=
attest
to
this. )
Bancroft's book talks about this phenome=n with amyloid, and
suggests
that
oxidation of the tissue (protein= due to exposure to air may be the
cause.
I'm guessing the it prob�ly applies to the precut microorganism
control
slides, too.
We only cut enough slides for 6 months, and date them. Other people
say
they
put their cut control slides in a slide box with a lid after dryi=,
and
then place the box in the refrig, as cold slows down the che=cal
change,
and the lid keeps the moving air off the slide. Other =ople say
that,
after
they dry the slide, they dip the slide in par�fin, to cover the
tissue,
so
that the air doesn't touch the tissue=ring the months before the
slide
is
used in a stain. Just 3 sugge=ions to stop this problem.
Peggy A. Wenk, HTL(ASCP)SLS
Beaumont=spital
Royal Oak, MI 48073
---=-
From: Amador, Amanda [8]aama...@ameripath.com
Sent: Wednesday, March 09, 2010:22 PM
To: [9]=sto...@lists.utsouthwestern.edu
Subject: [Histonet] Control Sl=es
Is there guidelines for how long special stains controls a= good
for
once
they are cut? We have spirochetes for our Stei=r that is from
2007
and
we are having issues.
=anda Amador, HT(ASCP)CM
AmeriPath | Histology Group Lead/Trainer=560 N Shadeland Ave,
Suite
A |
Indianapolis, IN 46219 | phon�17.275.8052 |
[10]aamador@=eripath.commailto:[11]aam�o...@ameripath.com; |
[12]www.AmeriPath.comhttp://www.ameripath.c=/;
___Histonet mailing list
[13]Histonet@lists.utsouthwestern.edu
[14]http://lists.utso=hwestern.edu/mailman/listinfo/histonet
__=___
Histonet mailing list
[15]Histonet@lists.utsouthwestern.edu[16]htt
p://lists.utsouthwestern.edu/mailman/listinfo/histonet
_=
Histonet mailing list
[17]Histonet@lists.utsouthwestern=du
[18]http://lists.utsouthwestern.edu/mailman/listinfo/histonet
References
1. file://localhost/tmp/3Dma 2. file://localhost/tmp/3Dmailto 3.
3Dmailto:aama...@ameripath.com;
4. 3Dmailto:histonet@lists.utsouthwestern.edu;
5. 3Dmailto:histonet-boun...@lists.utsouthwestern.edu;
6. 3Dmailto:histonet-bounc 7. 3Dmailto:histonet@lists.utsouth 8.
3Dmailto:aamador@ame 9.
[Histonet] RE: Beecher Instruments
Hello, There is a company that I just ran a few months ago. http://www.pathologydevices.com/TMArrayer.htm they have a manual arrayer that is running about 30,000.00. A little more than the Beecher Arrayer but it does offer some improvements. They are also able to sell the Needles that will fit into the Beecher instrument. I have spoken to Ron Gebing at Pathology Devices and he is responsive and reachable. I am very optimistic, He has many units in use in the U.S. and the world. Helen L. Fedor Tissue Microarray Lab, Manager Prostate Spore Lab, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 410.614.1660 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Barone, Carol Sent: Friday, March 04, 2011 3:56 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Beecher Instruments Importance: High Histonetters...It has been a longtime concern of mine that the only company offering TMA instrumentation was Beecher Instruments.and the wait list for those, was totally unacceptable. It came to my attention today, from a reliable source at a university that Beecher has gone out of business...( after buying Chemicon's arrayer...and taking away the only real alternative). I have heard from the same reliable source they may be purchased by a foreign investor...my prayer is that perhaps that could be Leica! Does anyone have more info on this? I wonder what this means for those who already have one.at least I am on the wait list! We have tried several manual systems from molds to punchesand there seems no real competition from these, which are difficult to control or expensive to use. What is the word out theresuggestions for us wait-ors. CB ___ 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] RE: Beecher Instruments
We too have tried the Quick Ray, and I think it is fine if you are not in the business. But if you only make a few arrays at a time then it is really wonderful I think for making small control block TMA's it is perfect. You can cut down the size of the recipient blocks to be able to make several out of each of these. The resultant blocks cut like butter. You don't lose a single section and you can cut them easily at 3 microns. Helen L. Fedor Tissue Microarray Lab, Manager Prostate Spore Lab, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 410.614.1660 From: Mark Tarango [mailto:marktara...@gmail.com] Sent: Friday, March 04, 2011 4:20 PM To: Helen Fedor Cc: Barone, Carol; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: Beecher Instruments There is also the Sakuro Quick-Ray System. I don't know if you've tried that already, but I've used it in the past and liked it. http://www.sakura-americas.com/products/tisstek-quickray.html The list price is $3950 for the whole kit and the recipient blocks are $68 each regardless of size (1 mm, 2 mm, 3 mm, or 5 mm). Mark On Fri, Mar 4, 2011 at 1:09 PM, Helen Fedor hfe...@jhmi.edumailto:hfe...@jhmi.edu wrote: Hello, There is a company that I just ran a few months ago. http://www.pathologydevices.com/TMArrayer.htm they have a manual arrayer that is running about 30,000.00. A little more than the Beecher Arrayer but it does offer some improvements. They are also able to sell the Needles that will fit into the Beecher instrument. I have spoken to Ron Gebing at Pathology Devices and he is responsive and reachable. I am very optimistic, He has many units in use in the U.S. and the world. Helen L. Fedor Tissue Microarray Lab, Manager Prostate Spore Lab, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 410.614.1660 -Original Message- From: histonet-boun...@lists.utsouthwestern.edumailto:histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edumailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Barone, Carol Sent: Friday, March 04, 2011 3:56 PM To: histonet@lists.utsouthwestern.edumailto:histonet@lists.utsouthwestern.edu Subject: [Histonet] Beecher Instruments Importance: High Histonetters...It has been a longtime concern of mine that the only company offering TMA instrumentation was Beecher Instruments.and the wait list for those, was totally unacceptable. It came to my attention today, from a reliable source at a university that Beecher has gone out of business...( after buying Chemicon's arrayer...and taking away the only real alternative). I have heard from the same reliable source they may be purchased by a foreign investor...my prayer is that perhaps that could be Leica! Does anyone have more info on this? I wonder what this means for those who already have one.at least I am on the wait list! We have tried several manual systems from molds to punchesand there seems no real competition from these, which are difficult to control or expensive to use. What is the word out theresuggestions for us wait-ors. CB ___ Histonet mailing list Histonet@lists.utsouthwestern.edumailto:Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edumailto: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] Section position on slides
Since automation is becoming more and more a part of all Histology labs the demands of placement of the tissue on the slides varies for different instruments. Stainers, coverslippers and now with slide scanning as well. So I do not believe that there is a silver bullet answer. Helen L. Fedor Tissue Microarray Lab, Manager Prostate Spore Lab, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 410.614.1660 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bartlett, Jeanine (CDC/OID/NCEZID) Sent: Wednesday, February 16, 2011 12:21 PM To: Tanya Ewing-Finchem; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Section position on slides I always try to center the section equally from sides and top-bottom as possible. This means measure from the bottom of the frosted edge as the top. The Artisan special stains system has a clip that attaches around the slide to allow reagents to pool onto the sections and incubate. If the section is too close to the sides then these areas do not stain adequately. Sometimes if the tissue section itself is very large this is unavoidable. With automated coverslippers you must also consider placement of tissue to allow for proper coverage. If I am cutting multiple unstained slides for subsequent testing I try to orient the tissue the same on each slide to facilitate the reading of these slides by the pathologist's Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartl...@cdc.hhs.gov -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Tanya Ewing-Finchem Sent: Wednesday, February 16, 2011 12:09 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Section position on slides I am trying to put together a training document around microtomy and sectioning and am finding it hard to find information around the placement of the actual sections on the slides. These are the objectives I am looking to answer. Is this information found in any publications? 1) Tissue / Section Placement: Are there published guidelines / documentation on precisely where you should place tissue sections on a 25mm x 75mm glass slide? Perhaps more importantly, where you should NOT place tissue (ie. x mm from the edge of the glass slide)? 2) Diagnosable Slide Staining Area: With automation becoming more widely used in IHC, are there published guidelines / documentation on the usable or diagnosable staining area on a 25mm x 75mm glass slide? For instance, would you define that as the area under a traditional coverslip? Would this be defined as the entire slide below the label? Or is this some distance from all the edges of the slide? With some automated systems, it is near impossible to get edge to edge staining. Is this acceptable? Thanks for any ideas. ___ 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] Charged slides
Hi, A question has come up regarding the different methods used to put a charge on the slide. We recently ordered some plus slides and the boxes they are packed in say silanized slides, but they say plus on the slide . We don't want to use these for our clients if they are not going to be getting the same results as the former plus slides that we were using. I was under the delusion that Plus slides somehow are magically charged without any coating process taking place. So does anyone know exactly how a plus slide gets its charge? Do they all get dipped in APES, or Polylysine? Thanks for your help. -- Helen ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] RE: Edge affect on TMA?
Here is a hint to keep your TMA from falling apart. After construction put the block face down on a glass slide a put in a 37degree C oven for 15-30 minutes (the paraffin should not be allowed to melt). Then take the block/slide out and gently compress with even pressure. (uneven pressure will cause the block to become misshapen). Cool the block at room temperature. You can repeat this process several times to help the cores adhere to the recipient block. The larger the core size, the more severe this problem is. One of the reasons that we choose not to use the 2.0 mm core is that this problem is more severe with this size core. Hope that will help you. Helen L. Fedor Tissue Microarray Lab, Manager Prostate Spore Lab, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 410.614.1660 From: IRENA SREBOTNIK KIRBIS [mailto:irena.kir...@hotmail.com] Sent: Wednesday, December 22, 2010 1:54 AM To: Helen Fedor; sette...@umdnj.edu; ahut...@dh.org; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Edge affect on TMA? how to prevent spots coming off during sectioning TM's? we do sometimes facing problems sectioning TM's, they're just falling apart? thanks for any hints Irena From: hfe...@jhmi.edu To: sette...@umdnj.edu; ahut...@dh.org; histonet@lists.utsouthwestern.edu Date: Tue, 21 Dec 2010 16:13:31 -0500 CC: Subject: [Histonet] RE: Edge affect on TMA? Hello, We do not usually see that. If we do its very rare because I can't think of any examples off hand. The usual problems are the spots came off or shifted/smushed, not an edge effect. Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Settembre, Dana Sent: Tuesday, December 21, 2010 3:00 PM To: 'Hutton, Allison'; histonet@lists.utsouthwestern.edu Subject: [Histonet] Edge affect on TMA? Does anyone experience edge affect on TMA? Dana Settembre ___ 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] RE: Edge affect on TMA?
Hello, We do not usually see that. If we do its very rare because I can't think of any examples off hand. The usual problems are the spots came off or shifted/smushed, not an edge effect. Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Settembre, Dana Sent: Tuesday, December 21, 2010 3:00 PM To: 'Hutton, Allison'; histonet@lists.utsouthwestern.edu Subject: [Histonet] Edge affect on TMA? Does anyone experience edge affect on TMA? Dana Settembre ___ 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] Pathology Grossing Manual
http://www.amazon.com/Surgical-Pathology-Dissection-Illustrated-ebook/dp/B000PY3QPM here is the one that we use. Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of tracz...@aol.com Sent: Wednesday, December 08, 2010 6:34 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Pathology Grossing Manual Amazon.com lists Manual of Surgical Pathology Gross Room Procedures by Juan Rosai. Of course it's not available. Does anyone know anything about this book? Is it worth tracking down? I need to update expand an existing lab manual and thought it would be helpful. Any ideas would be greatly appreciated. Dorothy MTA Histology LLC PO Box 602 Point Pleasant, NJ 08742 T:732-899-2912 ___ 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] unstained paraffin tissue slides storage--why cold?
The study that we did showed that the staining on freshly cut slides from a block stored at room temperature was in fact not as good as slides that were sectioned 5 years earlier and stored at -20. Therefore the blocks should be stored in the cold as well. The tissue is degrading in the block and on the slides and the cold does slow down the process. The fixation in lots of the tissue is not optimal. Fixing for 48 hours will definitely be better than the current fixation practices going on in most clinical labs, if what you want is the best tissue preservation possible. But most of us do not have that option. Helen L. Fedor Tissue Microarray Lab, Manager Prostate Spore Lab, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of sgoe...@xbiotech.com Sent: Thursday, November 04, 2010 11:21 AM To: Emily Sours Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] unstained paraffin tissue slides storage--why cold? -70 or -80 seems a little extreme to me, that's why I always just leave them in a normal freezer (-20). I think the main point of doing this from what I understand is so that the antigens stay viable. I know over time they can degrade and so your stain won't work with some antibodies. The weirdest part to me has always been that you don't have to store the blocks this way. So I think that was your question, if the blocks aren't stored in a freezer why store the slides? Won't the antigens in the blocks start to degrade as well? This is a question I would like to know the answer to as well... Sarah Goebel, B.A., HT (ASCP) Histotechnician XBiotech USA Inc. 8201 East Riverside Dr. Bldg 4 Suite 100 Austin, Texas 78744 (512)386-2907 Original Message Subject: Re: [Histonet] unstained paraffin tissue slides storage--why cold? From: Emily Sours talulahg...@gmail.com Date: Thu, November 04, 2010 7:07 am To: histonet@lists.utsouthwestern.edu Can I ask what the point of storing paraffin sections in freezing cold storage? They are wax sections, which never see any type of cold, so I don't understand the point of this. I do understand putting them at 4 degrees to prevent mold, but -80 seems excessive. We have kept our slides at room temperature for years and years, but these slides do not have an albumin coat (which I can see getting moldy), just a chemical coating. Fixing for paraffin and paraffin infiltration seems to keep antigens safe without refrigeration because it's so intense, but that's just conjecture on my part. Emily -- Outside of a dog, a book is man's best friend. Inside of a dog it's too dark to read. --Groucho Marx ___ 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: [Histonet] unstained paraffin tissue slides storage--why cold?
Hello, We have not published the paper yet . It is in progress and hope to get it out of the pile soon. Helen From: dusko trajkovic [mailto:dunat...@sbcglobal.net] Sent: Thursday, November 04, 2010 1:45 PM To: Morken, Tim; Helen Fedor Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] unstained paraffin tissue slides storage--why cold? I did an experiment about 10 or 12 years ago, where I cut the sections and stained them at various intervals: Day1, Day 15, 1 month, 3 months, 6 months, 1 year. Before staining: Set of slides were allowed to sit at room temp (RT). Set of slides were stored at 4C. Set of slides were vacuum sealed and stored at RT. Set of slides were vacuum sealed and stored at 4C. Slides stored at 4C consistently stained well, with a slight variance in staining after 1 year. Did not matter weather they were vacuum sealed or not. Slides left out at RT did not fare so well. Staining variability was noticed after 1 month. When the slides were stained after one year, signal was almost eliminated. Variability and loss of antigenicity was observed with the vacuum sealed slides as well. I did this experiment for just one antibody which was being extensively used at the time for one of our projects. It could be that other antibodies fare mach better under RT conditions, but why take a chance? We keep all of our control slides at 4C. Blocks are kept at RT. Thanks Dusko Trajkovic From: Morken, Tim timothy.mor...@ucsfmedctr.org To: Helen Fedor hfe...@jhmi.edu Cc: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Sent: Thu, November 4, 2010 9:09:16 AM Subject: RE: [Histonet] unstained paraffin tissue slides storage--why cold? Helen, did you write a paper from that study? Tim Morken Supervisor, Histology, IPOX UCSF Medical Center San Francisco, CA, USA -Original Message- From: histonet-boun...@lists.utsouthwestern.edumailto:histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edumailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Helen Fedor Sent: Thursday, November 04, 2010 8:56 AM To: sgoe...@xbiotech.commailto:sgoe...@xbiotech.com; Emily Sours Cc: histonet@lists.utsouthwestern.edumailto:histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] unstained paraffin tissue slides storage--why cold? The study that we did showed that the staining on freshly cut slides from a block stored at room temperature was in fact not as good as slides that were sectioned 5 years earlier and stored at -20. Therefore the blocks should be stored in the cold as well. The tissue is degrading in the block and on the slides and the cold does slow down the process. The fixation in lots of the tissue is not optimal. Fixing for 48 hours will definitely be better than the current fixation practices going on in most clinical labs, if what you want is the best tissue preservation possible. But most of us do not have that option. Helen L. Fedor Tissue Microarray Lab, Manager Prostate Spore Lab, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 -Original Message- From: histonet-boun...@lists.utsouthwestern.edumailto:histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edumailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of sgoe...@xbiotech.commailto:sgoe...@xbiotech.com Sent: Thursday, November 04, 2010 11:21 AM To: Emily Sours Cc: histonet@lists.utsouthwestern.edumailto:histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] unstained paraffin tissue slides storage--why cold? -70 or -80 seems a little extreme to me, that's why I always just leave them in a normal freezer (-20). I think the main point of doing this from what I understand is so that the antigens stay viable. I know over time they can degrade and so your stain won't work with some antibodies. The weirdest part to me has always been that you don't have to store the blocks this way. So I think that was your question, if the blocks aren't stored in a freezer why store the slides? Won't the antigens in the blocks start to degrade as well? This is a question I would like to know the answer to as well... Sarah Goebel, B.A., HT (ASCP) Histotechnician XBiotech USA Inc. 8201 East Riverside Dr. Bldg 4 Suite 100 Austin, Texas 78744 (512)386-2907 Original Message Subject: Re: [Histonet] unstained paraffin tissue slides storage--why cold? From: Emily Sours talulahg...@gmail.commailto:talulahg...@gmail.com Date: Thu, November 04, 2010 7:07 am To: histonet@lists.utsouthwestern.edumailto:histonet@lists.utsouthwestern.edu Can I ask what the point of storing paraffin sections in freezing cold storage? They are wax sections, which never see any type of cold, so I don't understand the point of this. I do understand putting them at 4 degrees to prevent mold, but -80 seems excessive. We have kept
RE: [Histonet] unstained paraffin tissue slides storage
Hello, We have been storing our slides in very small Ziploc bags at -20 and find that this method works fairly well. We have done a study(Berez et.al in process) and slides stored in this fashion for 5 years stain better than freshly cut slides from the same blocks that have been stored at room temperature. Helen L. Fedor Tissue Microarray Lab, Manager Prostate Spore Lab, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 410.614.1660 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of sgoe...@xbiotech.com Sent: Wednesday, November 03, 2010 4:26 PM To: Pop Elena Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] unstained paraffin tissue slides storage Plain slide boxes is ok. I think you can store them for up to a year in a fridg. (4 degrees), but I usually pu them in a freezer (-20). Sarah Goebel, B.A., HT (ASCP) Histotechnician XBiotech USA Inc. 8201 East Riverside Dr. Bldg 4 Suite 100 Austin, Texas 78744 (512)386-2907 Original Message Subject: [Histonet] unstained paraffin tissue slides storage From: Pop Elena med_l...@yahoo.com Date: Wed, November 03, 2010 1:21 pm To: histonet@lists.utsouthwestern.edu Hello, I found here a few disscussions regarding the storage of tissue slides but I did not find a clear answer to the questions I have. I would really appreciate an answer from anybody that has experience with this. I need to store for long term a bunch of unstained tissue slides for the purpose of doing immunostaining even in a few years from now on. Unfortunatelly they were stored for about 3 years at room temperature. What it is usually recomended: to store them at -20 degrees Celsius? If yes, is it OK to store them in the regular 100 slides boxes? And when you need to start an immunostaining just take them out of the freezer and let them at room temp for a while before starting the stain or what procedure do you use? I heard some labs keep them in nitrogen gas containers. Do you have any info about this? Any imput is appreciated! 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Block release
Hello, Our institution does release blocks, the requirement is that a patient consent is signed and we have an MTA (Material Transfer Agreement) For the study and appropriate IRB's in place for the study. It is a lot to set up, but after this is done for one study any following studies are easier to handle. Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Bernice Frederick Sent: Monday, October 18, 2010 11:04 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Block release Everyone, I have been asked to post a query as to what your institution does in terms of releasing blocks for oncology clinical trials. Please respond as it is important to us as we receive a lot of those blocks here at Northwestern (policies, procedures, alternative submissions etc) If you are in Australia, Ireland, Canada, Puerto Rico or Peru please also answer (though I sort of know already) as we do get blocks from you also. Thanks Bernice Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility ECOGPCO-RL 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 ___ 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] Cutting Microarrays
Hello, We have trouble with the larger size punches as well. In order to get the cores to stick to the size try warming the block in a oven at 37 degrees for 15 minutes, put the block face down on a clean slide. (providing the melting temperature of the paraffin is about 57 degrees). After 15 minutes remove the block/slide complex and gently press down on the block, then allow the block to cool. Repeat the process 3 or 4 times. We have also had some success by sliding the block face over the heating unit of the paraffin embedder. But this only works for a few sections and then you get the same problem back. Best Regards, Helen L. Fedor Tissue Microarray Lab, Manager Prostate Tissue Spore Lab, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 410.614.1660 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of louise renton Sent: Monday, September 20, 2010 5:51 AM To: Rae Staskiewicz; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Cutting Microarrays hi, this sounds like the tissue was too cool when embedded in the wax - so it has not made a nice homogenous block that cuts as one. Can u re-embed? On Mon, Sep 20, 2010 at 10:54 AM, Rae Staskiewicz raest...@grics.netwrote: Good Morning All, I am having some issues when cutting a microarray containing 4mm punches. The punches are rolling and separating from the paraffin during cutting. Anyone have any tips or ideas?? Rae Staskiewicz MMCI ___ 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 +27 11 717 2298 (tel fax) 073 5574456 (emergencies only) There are nights when the wolves are silent and only the moon howls. George Carlin No trees were killed in the sending of this message. However, many electrons were terribly inconvenienced. ___ 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] Cutting Microarrays
Hello, We have trouble with the larger size punches as well. In order to get the cores to stick to the size try warming the block in a oven at 37 degrees for 15 minutes, put the block face down on a clean slide. (providing the melting temperature of the paraffin is about 57 degrees). After 15 minutes remove the block/slide complex and gently press down on the block, then allow the block to cool. Repeat the process 3 or 4 times. We have also had some success by sliding the block face over the heating unit of the paraffin embedder. But this only works for a few sections and then you get the same problem back. Best Regards, Helen L. Fedor Tissue Microarray Lab, Manager Prostate Tissue Spore Lab, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 410.614.1660 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of louise renton Sent: Monday, September 20, 2010 5:51 AM To: Rae Staskiewicz; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Cutting Microarrays hi, this sounds like the tissue was too cool when embedded in the wax - so it has not made a nice homogenous block that cuts as one. Can u re-embed? On Mon, Sep 20, 2010 at 10:54 AM, Rae Staskiewicz raest...@grics.netwrote: Good Morning All, I am having some issues when cutting a microarray containing 4mm punches. The punches are rolling and separating from the paraffin during cutting. Anyone have any tips or ideas?? Rae Staskiewicz MMCI ___ 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 +27 11 717 2298 (tel fax) 073 5574456 (emergencies only) There are nights when the wolves are silent and only the moon howls. George Carlin No trees were killed in the sending of this message. However, many electrons were terribly inconvenienced. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Chilling trays
You can try disecting trays. I just googled discting tray, looks like listing price is 17.00. they are the perfect size and shape for freezing. and keep blocks organized. Helen Fedor Johns Hopkins University TMA and Prostate Spore Lab Manager JHU SOM Uro/Pathology 600 N. Wolfe Street, Marburg Room 406 Baltimore MD, 21287-7065 410-614-1660 / Fax 410-614-3695 Pager 410-283-3419 From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Breeden, Sara [sbree...@nmda.nmsu.edu] Sent: Thursday, July 01, 2010 10:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Chilling trays I went to eBay and found a set of three (total $12.00) stainless steel food prep trays, roughly 9x5x2 (LxWxH). I fill them with DI water and freeze them. I also inherited a larger stainless steel container that's 12.5x10.5x4.5 - same deal, DI water and freeze. I face blocks, put them on the ice and add a little DI water and soak for 10-15 minutes - blocks cut like a dream! They stay frozen for a l - o - n - g time! Try a local restaurant supply. Sally Breeden, HT(ASCP) Veterinary Diagnostic Services New Mexico Department of Agriculture 700 Camino de Salud NE Albuquerque, NM 87108 505-841-2576 ___ 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] RE: Prolonged FFPE slide and block storage
Hello, We have done a study and found that storage in small zip-loc bags at -20 is an inexpensive and convenient way to store slides. In fact the slides stored for 5 years at -20 stain better than freshly cut sections from the same blocks. This was also published recently by others. Dr Rimm at Yale does have the storage under nitrogen gas system that is available commercially. Helen L. Fedor Tissue Microarray Lab, Manager Prostate Spore Lab, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 410.614.1660 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Saturday, May 15, 2010 9:36 AM To: SamuelPerry; histonet@lists.utsouthwestern.edu; TimMorken Subject: Re: [Histonet] RE: Prolonged FFPE slide and block storage Inexpensive long term anti oxidizing storage? Immerse them in mineral oil! René J. --- On Fri, 5/14/10, Morken, Tim timothy.mor...@ucsfmedctr.org wrote: From: Morken, Tim timothy.mor...@ucsfmedctr.org Subject: [Histonet] RE: Prolonged FFPE slide and block storage To: Perry, Samuel samuel_pe...@dfci.harvard.edu, histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Date: Friday, May 14, 2010, 5:05 PM I have a paper from Yale several years ago about their tissue array facility. They found that storage in nitrogen helped a lot. I will try to find it, but you may be able to google it faster. Tim Morken Supervisor, Histology / IPOX UCSF Medical Center San Francisco, CA -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Perry, Samuel Sent: Friday, May 14, 2010 1:16 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Prolonged FFPE slide and block storage Hi All, We have a growing need for storing slides. They need to be stored in a way that they won't become oxidized and loose their antigenicity, since we use them for IHC. Any suggestions for inexpensive methods for prolonged storage of FFPE slides and blocks is greatly appreciated. Thanks! Sam Perry Research Technician Dana-Farber Cancer Institute Boston, MA 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Cover glass
Hello, Our lab does IHC staining in house and have had zero problems with our Fisher brand cover slips until recently. We've tried both Fisher and Corning cover slips and what appears to be dust or imperfections in the glass are found on both brands. This is creating false positives for us, because when the positive staining is low it is difficult to impossible to tell the imperfections from the staining itself. Any advice will be welcome. -- Helen ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Akemi - Urgent!
I also receive one of the scam emails. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Breeden, Sara Sent: Thursday, November 12, 2009 9:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Akemi - Urgent! Akemi Allison-Tacha - email me immediately, please. I think you are being scammed and I don't want to email you directly. Sally Breeden, HT(ASCP) NM Dept. of Agriculture Veterinary Diagnostic Services PO Box 4700 Albuquerque, NM 87106 505-841-2576 ___ 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] withdrawal
To subscribe or unsubscribe to the list or change your subscription to the daily digest mode etc. you need to go to: http://lists.utsouthwestern.edu/mailman/listinfo/histonet or email: histonet-reque...@lists.utsouthwestern.edu with the word help (without quotation marks) in the subject line or body of the message. Instructions will be returned to you by email. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Roger Moretz Sent: Wednesday, October 21, 2009 5:38 PM To: ji...@bme.ogi.edu; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] withdrawal The correct request is to unsubscribe but since you forgot the original information you received when you first subscribed or have failed to read any of the multitude of recent messages about unsubscribing, I'll guess you'll just have to suffer along with the rest of us in getting too many messages. Roger Moretz, Ph.D. - Original Message From: ji...@bme.ogi.edu ji...@bme.ogi.edu To: histonet@lists.utsouthwestern.edu Sent: Wed, October 21, 2009 4:26:58 PM Subject: [Histonet] withdrawal Hi Please help me to withdrawal from the histonet. I am frustrated to see a huge amount email each day. Thanks yali ___ 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] RE: re: control slide storage
Hello, We have a project that has worked on this topic using TMA's and have found that storing unstained, and unbaked slides at -20 enhances the preservation of antigenicity of the samples. It is not yet published but is getting written up. TMA slides that were stored for 5 years at -20 in very small Ziploc bags were compared to freshly cut TMA slides of the same block. Surprisingly some of the freezer slides tissue stained better then the freshly cut slides. So we need to start to store our tissue blocks in the cold as well. -- Helen L. Fedor Tissue Microarray Lab, Manager Prostate Spore Lab, Manager Johns Hopkins University 600 N. Wolfe St, | Marburg Room 406 Baltimore, MD | 21287-7065 410.614.1660 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Renko, Heather D. Sent: Thursday, June 25, 2009 4:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] re: control slide storage Anyone have any published literature for reference on storage control; refrigerator vs. room temp vs. freezer. I have a friend asking for this and would also would love to get this information for my own reference I searched the archives but, came up with only one publication. Thank you in advance. Heather D. Renko, Histology Supervisor OSF Saint Anthony Medical Center 5666 East State Street Rockford, Illinois 61108 Direct: 815-395-5410 heather.d.re...@osfhealthcare.org == The information in this message is confidential and may be legally privileged. Access to this message by anyone other than the addressee is not authorized. If you are not the intended recipient, or an agent of the intended recipient, any disclosure, copying, or distribution of the message or any action or omission taken by you in reliance on it, is prohibited and may be unlawful. If you have received this message in error, please contact the sender immediately 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Full Mount Immunos
I believe that Brain Research Labs has large + slides. Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Metzger, Kenneth Sent: Thursday, March 05, 2009 10:07 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Full Mount Immunos Does anyone know of any companies producing charged full mount slides? I have a pathologist that wants immunos done on full mount prostates and the tissue keeps falling off. I have looked, but cannot find any charged full mount slides. Any help would be appreciated!! Kenneth G Metzger HTL(ASCP) ARUP Labs Histology Supervisor 500 Chipeta Way Salt Lake City, Utah 84108-1221 kenneth.metz...@aruplab.com (801) 583-2787 x 3101 - -- The information transmitted by this e-mail and any included attachments are from ARUP Laboratories and are intended only for the recipient. The information contained in this message is confidential and may constitute inside or non-public information under international, federal, or state securities laws, or protected health information and is intended only for the use of the recipient. Unauthorized forwarding, printing, copying, distributing, or use of such information is strictly prohibited and may be unlawful. If you are not the intended recipient, please promptly delete this e-mail and notify the sender of the delivery error or you may call ARUP Laboratories Compliance Hot Line in Salt Lake City, Utah USA at (+1 (800) 522-2787 ext. 2100 ___ 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] Pathos animal tissue processing
Hello, Another question, Is anyone using the Pathos system to process animal tissues? I would really appreciate any comments good/ and or bad on this topic. Best regards, Helen From: Helen Fedor Sent: Wednesday, February 18, 2009 5:33 PM To: 'histonet@lists.utsouthwestern.edu' Subject: Pathos Processing tissue problems Hello, Our Surgical Pathology Department has been using the Pathos processor. I work in research and we have begun to have issues with the tissue falling off of the slide after pretreatment for IHC. Has anyone else noticed this happening? The tissue is much harder in the block than the standard method of processing. Is this normal? Does that have anything to do with the tissue falling off the slides? Thanks in advance. Helen L. Fedor Comprehensive Tissue Services Center Manager Prostate Spore Lab Manager 600 N. Wolfe St, Marburg Room 406 Baltimore, MD 21287-7065 410-614-1664 email:hfe...@jhmi.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Pathos Processing tissue problems
Hello, Our Surgical Pathology Department has been using the Pathos processor. I work in research and we have begun to have issues with the tissue falling off of the slide after pretreatment for IHC. Has anyone else noticed this happening? The tissue is much harder in the block than the standard method of processing. Is this normal? Does that have anything to do with the tissue falling off the slides? Thanks in advance. Helen L. Fedor Comprehensive Tissue Services Center Manager Prostate Spore Lab Manager 600 N. Wolfe St, Marburg Room 406 Baltimore, MD 21287-7065 410-614-1664 email:hfe...@jhmi.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Cryostat safety question
Hello, I had previously sent this but only to the person who asked the question, I think it this works beautifully so decided to resend to the list. we have a great way to remove the specimens for the chucks. We have a 500cc plastic container with a lid that we have cut an X into the center. We put warm water in the container and then just put the chuck stem into the X. Within 10 seconds the block can be removed. Still frozen. We just keep the plastic container at the bench and keep reusing the same one. The water level does need to come all the way up to the lid, But the chuck never gets wet. We do the sealing of the tissue with a tiny amount of OCT to the surface of the still frozen block, while it is still in the cryostat as well. Helen -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Ingles Claire Sent: Wednesday, January 07, 2009 7:38 PM To: Histonet Cc: mari.ann.mailh...@leica-microsystems.com Subject: RE: [Histonet] Cryostat safety question Andrea: I work in a Mohs clinic where all we cut is frozen skin sections. Needless to say, we don't have 50 chucks laying around... In the morning before clinic starts we put a layer of freezing medium on chucks and put them in the cryostat to freeze. When we get specimens, we add another drop or so to the already frozen 'button' and immediately embed the tissue in it. We usually add another small drop on top after it has begun to freeze, to cover the specimen completely. Cut as normal when frozen. After done cutting all you have to do is use a forceps or other blunt object and pop the bit with the specimen in it away from the 'button' and return the chuck to the cryostat and it can be reused the rest of the day. The specimen is therefore still frozen for storage, and it has a quicker TAT. Plus you won't need nearly so many chucks, as they can be recycled almost as soon as you are done cutting. I usually keep 6-8 'buttons' in my cryostat, and our clinic can process up to 50 separate specimens a day. A word of caution. If your work area is humid sometimes a thin layer of frost can form on the surface of the 'button' and when you attempt to take sections the bit with the tissue will pop off the 'button'. All you need to do is add another drop of medium to the button and 'glue' the two back together. If you are going a while between cutting sessions, I usually store my 'buttons' upside(mountant side) down on one of the cryostat surfaces. It doesn't seem to develop the frost layer. Useful if you have tiny specimens. Hope my verbose explanation is helpful. Feel free to e-mail if you have any questions or are confused about my explanation. Claire Ingles Madison WI From: histonet-boun...@lists.utsouthwestern.edu on behalf of Andrea Hooper Sent: Wed 1/7/2009 5:40 PM To: Histonet Cc: mari.ann.mailh...@leica-microsystems.com Subject: [Histonet] Cryostat safety question The discussion on microtome safety begs me to ask a cryostat question We have a Leica CM3050 cryostat and love it! How are people (and perhaps only those in research do this) removing their tissue from the chucks for future use? We often just section a few slides worth then put the block at -80 deg C for future studies. Needless to say, it's the most dangerous part of our day. So what are your suggestions for removing tissue from a chuck (and melting it isn't really a viable option)? Thanks in advance, 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 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
