RE: SPAM-LOW: Reponses to Merced and Richard Re: [Histonet] cryojane tape transfer system
I guess I did not realize what ongoing discussion my comment about the CryoJane tape transfer would spark! Thanks to the replies I have received in private and in public, I see that it indeed helps one make lovely bone sections. In the past 9 years that I have cryosectioned, I have done a range of soft tissues, but never bone. CryoJane had been pushed on me a time or two by a rep to use on my (non-bone) tissues. I discovered, however, that it was not necessary on my (non-bone) tissues. I was never informed of using it on bone (until now!), never even crossed my mind, since I'd never done bone. So, I'm sorry for anyone who may have been offended by my question about the use of the CryoJane tape transfer. I indeed figured I had to be missing something! :-) Regards, Merced --On Sunday, December 06, 2009 2:31 PM -0700 Patsy Ruegg pru...@ihctech.net wrote: Here, here Gayle, I have a picture on my website of a calcified horse carpal bone I cut using the Instrumedics tape transfer system, I bet no one would have been able to do that without using the tape. www.ihctech.net Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. Ste.215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 www.ihctech.net www.ihcrg.org -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of gayle callis Sent: Monday, November 23, 2009 10:21 AM To: 'Histonet' Cc: emmanuel.mi...@leica-microsystems.com Subject: SPAM-LOW: Reponses to Merced and Richard Re: [Histonet] cryojane tape transfer system You wrote: Unless I'm missing something, I don't understand why people use this tape? It seems like a marketing gimmick to me...ol' fashion' melting of sections onto slides works perfectly for us... ? Regards, Merced Merced, Yes, you are missing something. If you have ever tried to cryosection undecalcified bone or extremely difficult tissues that simply will not result in ol' fashion' melting onto a slide , then you would understand why people use this unique cryosectioning system. It is not some marketing gimmick but an unique instrument helping many laboratories obtain frozen sections that otherwise are scrunched up, shattered, and destroyed. I suggest you go to the Instrumedics website or www.alphelys.com for a superb slide show and learn how this instrument works before making assumptions about a technology that serves many of us more than well. A happy, informed user of the Cryojane Gayle M. Callis HTL/HT/MT(ASCP) Bozeman MT As for what Richard wrote: Hi Everybody, I was hoping to get some advice - I'm cryosectioning plant tissues and transferring sections to slides using the Cryojane system. However, i'm having problems in transferring the sections without them falling apart during the tape transfer. I'm fixing my tissue for 24 hours in ethanol:acetic acid (3:1), embedding in O.C.T, snap-freezing and then sectioning at between 2 and 14 microns. The sections seem to be ok but whenever i remove the adhesive tape from the slide a large part of the tissue is removed with it. As a result I lose the majority of my section. I've tried using both 1x and 1/2x slides (CFSA adhesive slides from instrumedics) but neither have given satisfactory results. Does anyone have any suggestions as to how i could reduce the loss of tissue? Any advice would be much appreciated. Thanks Richard Dear Richard, I could be the fixative you are using that causes the problem. If the fixative contains alcohol, the alcohol acts as antifreeze when you try to snap freeze a tissue, animal or plant. The alcohol may cause problems with how the pink tape sticks to the face of the plant tissue, and allows them to fall apart during the tape transfer. If you rinse away the fixative, then you should cryoprotect the fixed plant tissue with 30% sucrose before snap freezing. vbThis will remove the alcohol. If cryoprotection causes problems with the final staining results, then try unfixed plant tissue, snap freeze, Cryojane tape transfer the section and then fix the transferred plant section in your favorite fixative. You may have to optimize the time in fixative though. Other suggestions: Do a double UV flash, but wait for 15 to 20 seconds between flashes. You must allow the UV light source (capacitor) build up enough charge to work properly. This double flash seems to help polymerize the coating more thoroughly, and the section should transfer better. Also, the tape must be removed at an angle across the slide, very slowly, and inside the cryostat (I am sure you probably do this already.) Also, try the 4X slide if you still have problems with 1/2X and 1X slides. You might ask Leica to send you a few to try before investing in a whole box of these. Contact Emmanuel Mineo, Intrumedics Product Manager emmanuel.mi...@leica-microsystems.com for the 4X slides. Manny is a nice gentleman who has worked with
[Histonet] Cutting GI biopsies with multiple levels
Histonet, One of our pathologists is looking at a method for cutting GI biopsies that differs in complexity from the way we currently do them. Currently we cut just 2 levels at 4 microns thick, 40 microns apart and only showing 2 sections on each level. In the new plan, if there were a maximum of 5 small pieces per block, the idea is that we would cut 10 serial sections at 4 microns thickness, split the ribbons in half on the waterbath, and then put 5 sections vertically on 1 side of the slide and the other 5 sections vertically on the other side of the slide. We would then face in 40 microns and repeat the process for a second slide for a total of 2 slides, 20 sections of tissue, 120 microns between the first and the last section. I am happy to do this if our pathologists believe that it would improve their diagnosis. Since we do about 400 biopsies like this a month, I would like to look at our workload to see how to change our staffing with this method. I would like to get an idea of the difference in time between the 2 methods. Does anyone else do a similar method and would be willing to share their experiences? I am primarily interested in the amount of time it would take an experienced tech to cut a biopsy like this. We have tried it out, but our techs are far from experienced with this method. Thanks for your time. -- Patrick Laurie HT(ASCP)QIHC Histology Supervisor CellNetix Pathology Laboratories 1124 Columbia Street, Suite 200 Seattle, WA 98104 plau...@cellnetix.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Histology Jobs
Dear Histonet Subscribers, I am a one of the founders of a Healthcare Recruiting firm that specializes in placing Lab Professionals. We work exclusively on permanent positions and have clients across the country. We are completely free of charge to candidates and are currently working on numerous Histology positions. Our clients often assist with relocation expenses. Below is a list of some of the Histology opportunities we are currently working on: 1) New York City - Histotechnologist -3rd shift 2) New York City - Histology Supervisor - 1st Shift 3) Oklahoma - Histotechnologist - 1st shift 4) Georgia (SE part of the state) - Histology Supervisor - 1st shift 5) Georgia (SE part of the state) - Histotechnologist - 2 positions open 6) California (Bay Area) - Pathology Manager - 1st shift 7) California (Los Angeles) - Histology Supervisor - 1st shift 8) California (San Diego) - Sr. Histotechnologist - 1st shift 9) Massachusetts (Boston) - Histotechnologist - 1st shift Contact me if you're interested in learning more about any of these opportunities or if you'd like like me to tailor a specific search for you. We work on positions at all levels and cover the entire US. Please also send me an updated resume and let me know what the best way to reach you is. To view some additional opportunities please visit our website at www.ka-recruiting.com. Sincerely, KC Carpenter K.A. Recruiting, Inc. 10 Post Office Square, 8th Floor South Boston, MA 02109 P: (617) 692-2949 F: (617) 507-8009 k...@ka-recruiting.com www.ka-recruiting.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Cutting GI biopsies with multiple levels
It takes a while to get experienced, but it doubles, sometimes triples the time. Joyce -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Pat Laurie Sent: Wednesday, December 09, 2009 13:34 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cutting GI biopsies with multiple levels Histonet, One of our pathologists is looking at a method for cutting GI biopsies that differs in complexity from the way we currently do them. Currently we cut just 2 levels at 4 microns thick, 40 microns apart and only showing 2 sections on each level. In the new plan, if there were a maximum of 5 small pieces per block, the idea is that we would cut 10 serial sections at 4 microns thickness, split the ribbons in half on the waterbath, and then put 5 sections vertically on 1 side of the slide and the other 5 sections vertically on the other side of the slide. We would then face in 40 microns and repeat the process for a second slide for a total of 2 slides, 20 sections of tissue, 120 microns between the first and the last section. I am happy to do this if our pathologists believe that it would improve their diagnosis. Since we do about 400 biopsies like this a month, I would like to look at our workload to see how to change our staffing with this method. I would like to get an idea of the difference in time between the 2 methods. Does anyone else do a similar method and would be willing to share their experiences? I am primarily interested in the amount of time it would take an experienced tech to cut a biopsy like this. We have tried it out, but our techs are far from experienced with this method. Thanks for your time. -- Patrick Laurie HT(ASCP)QIHC Histology Supervisor CellNetix Pathology Laboratories 1124 Columbia Street, Suite 200 Seattle, WA 98104 plau...@cellnetix.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This email, 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 reply to the sender that you have received the message in error, then delete this message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] was Overstaining - Mayers HE
On the subject of protocols being passed down from person to person, lab to lab, etc., a freshly minted PhD came to me once looking for some NaH with which to make buffer. I explained that there was no such chemical as NaH but she insisted: here is the protocol. A simple typographical error had left out the O in NaOH. Because she had a full time tech at her beck and call throughout her PhD training she had never learned how to make simple solutions. Sad but true. Geoff John Kiernan wrote: This is a typical example of the informal protocols that get passed on from generation to generation of graduate students, postdocs and technicians in research labs at universities. The original was probably written by someone who knew how to do HE staining, but on differently fixed tissues, and certainly on thinner sections. It apears to be for individual slides, because 10 seconds in each of the two 95% and 100% ethanols would be effective only with vigorous agitation in a large excess of fluid. The tissue is almost unfixed, unless Soak in 0.4% paraformaldehyde means leave it ovenight or longer in 4% formaldehyde. Researchers otherwise educated to the highest levels in such difficult disciplines as molecular biology and neuroscience regularly write phrases like 4% paraformaldehyde, thereby advertising their profound ignorance about fixation, which is the procedure that has the greatest effect on the appearance of anything dead that's examined with a microscope, especially if stains or histochemical methods are to be used. (I apologise for the length of the preceding sentence, but not for its punctuation, which is correct in British but not in American English usage. Check it out with Lynne Truss!) The sucrose cycle step, with no times or instructions about floating and sinking, is probably local jargon from a lab where small animals' brains are minimally fixed and cryoprotected before cutting thick (50-100um) frozen sections, to be stained free-floating. That's not an HE job! You are working with a thin skeletal muscle (rat's gastrocnemius). If your Mayer's haemalum is a bought solution, it is intended for use in hospital labs, on paraffin sections about 5um thick. In a research setting you may need to make changes. Haemalum (Mayer's or anyone else's, correctly used) should stain cell nuclei blue and very little else. An important part of HE staining is looking at the wet section with a microscope to check for adequate and selective nuclear coloration. In skeletal muscle the nuclei are small, so the haemalum-stained section is very pale blue to the unaided eye. With alcoholic eosin (as in your method) it's not so easy to control the staining with microscopic control, but it will probably be OK if the section is light pink. Some people like their eosin darker; it's largely a matter of taste unless you need to distinguish between different eosinophilic components on the basis of hue. John Kiernan Anatomy, UWO London, Canada = = = -Original Message - From: Josephine Garcia j...@u.washington.edu Date: Monday, December 7, 2009 11:43 Subject: [Histonet] Overstaining - Mayers HE To: histonet@lists.utsouthwestern.edu Hi all, My (frozen-section, fixed) slides are coming out much too dark (overstainedpurple) and I'm not sure why. They are 15-20 micrometer slices of rat gastrocnemius muscle. Can someone please look over our current protocol and tell me what I'm doing wrong? Thanks! Here it is: 1. Perfuse animal with 4% paraformaldehyde fixative. 2. Soak in 0.4% paraformaldehyde 3. Sucrose cycle (5% rinse, 10%, 20%, 30% soak) 4. Embed in OCT, Frozen sections (15-20 micrometers) 5. Let dry for 15-30 min 6. Stain as follows: - Distilled H2O (quick dip) - Mayer's Hematoxylin - 1min (originally we were dipping for 5- 10 minutes. I slowly reduced the time to 2min, then 1min, then 30s... still overstained!)- Running lukewarm tap water - drain and continuously fill - 15min or until water runs clear - Distilled H2O (quick dip) - 80% EtOH - 1-2min - Eosin - 2 min - 95% EtOH I - 10sec - 95% EtOH II - 10sec - 100% EtOH I - 10sec - 100% EtOH II - 10sec - Xylene I - 2min - Xylene II - 2min 7. Coverslip and let dry overnight ___ 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 -- -- ** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583 mcaul...@umdnj.edu ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [HISTONET] fixation and storage of tissue for TEM
Fix in buffered glutaraldehyde for 2 hours, overnight is acceptable. After multiple rinses, store in buffer in the refrigerator. Change the buffer once a week, there are things that will grow in cacodylate buffer. You may complete osmication if you want, then rinse and store in buffer. I would not store the tissue indefinitely in anything. Do NOT store the tissue in 70% ethanol, either before or after osmium, ethanol has been shown to extract cytoplasm from fixed tissues. The texts John Kiernan recommends are excellent. Geoff Nicholas David Evans wrote: Dear all, I would like to fix and preserve mouse skin tissue for processing for transmission electron microscopy (TEM) at a later date. I was wondering whether I can store tissue following fixation overnight in 2.5% gluteraldehyde in 0.1M sodium cacodylate buffer and, if so, whether it can be stored indefinitely in this buffer (or whether I need to treat with OsO4 and store at 70% ethanol)? Thanks and best wishes Nick ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- -- ** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583 mcaul...@umdnj.edu ** ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Help! Losing sections from Superfrost Plus Slides
Hi all, We have suddenly started losing tissue sections from our Superfrost Plus Slides. Our students have been cutting fixed, frozen, cryosections (20um) and thaw-mounting these onto Superfrost Plus slides. We have suddenly started losing lots of sections from these slides (again!!). The tissue is small - ie cross sections of frog aorta and longitudinal sections of nerves, so any lose of adhesion results in total loss of the tissue. The odd thing is that she is not losing every section on every slide, but half to 3/4 of the sections are falling off within the first rinse. The sections are from the same tissue block on the same slide while some falls and others don't. We are at a lose as to what we can do to rescue these sections. Does anyone know if there is any way to coat the slides in some solution with the tissue on them to help improve the adhesion without losing the ability to do immunofluorescence? Any further advice on cutting / drying protocols are welcomed. This keeps happening and the inconsistency of it has us so frustrated with this that we are thinking of going back to subbing our own slides. Thanks for your help! Angelina -- ~~o~~o~~o~~o~~o~~o~~o~~o~~o~~o Angelina Y. Fong, Ph.D. Department of Zoology Biological Sciences Building 6270 University Boulevard University of British Columbia Vancouver, BC, V6T 1Z4 Canada Ph: (604) 822-5799 Fax: (604) 822-2416 Email: f...@zoology.ubc.ca ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Help! Losing sections from Superfrost Plus Slides
Angelina, I have seen this post a few times before and sometimes the problem was the drying technique or making sure your water bath is clean and free of any debris. I also remember hearing about someone pre-treating their slides with a Trilogy (EDTA) buffer in a pressure cooker. Good Luck! *Matthew Semovoski* Sales Manager Gorilla Scientific Corporation 1-866-435-4977 www.gorillascientific.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Cutting GI biopsies with multiple levels
An experienced histotach can cut regular blocks at a rate of 24/hour. What you describe will reduce the productivity to 8 to 10 blocks/hour. René J. --- On Wed, 12/9/09, Weems, Joyce jwe...@sjha.org wrote: From: Weems, Joyce jwe...@sjha.org Subject: RE: [Histonet] Cutting GI biopsies with multiple levels To: Pat Laurie foreig...@gmail.com, histonet@lists.utsouthwestern.edu Date: Wednesday, December 9, 2009, 1:55 PM It takes a while to get experienced, but it doubles, sometimes triples the time. Joyce -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Pat Laurie Sent: Wednesday, December 09, 2009 13:34 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cutting GI biopsies with multiple levels Histonet, One of our pathologists is looking at a method for cutting GI biopsies that differs in complexity from the way we currently do them. Currently we cut just 2 levels at 4 microns thick, 40 microns apart and only showing 2 sections on each level. In the new plan, if there were a maximum of 5 small pieces per block, the idea is that we would cut 10 serial sections at 4 microns thickness, split the ribbons in half on the waterbath, and then put 5 sections vertically on 1 side of the slide and the other 5 sections vertically on the other side of the slide. We would then face in 40 microns and repeat the process for a second slide for a total of 2 slides, 20 sections of tissue, 120 microns between the first and the last section. I am happy to do this if our pathologists believe that it would improve their diagnosis. Since we do about 400 biopsies like this a month, I would like to look at our workload to see how to change our staffing with this method. I would like to get an idea of the difference in time between the 2 methods. Does anyone else do a similar method and would be willing to share their experiences? I am primarily interested in the amount of time it would take an experienced tech to cut a biopsy like this. We have tried it out, but our techs are far from experienced with this method. Thanks for your time. -- Patrick Laurie HT(ASCP)QIHC Histology Supervisor CellNetix Pathology Laboratories 1124 Columbia Street, Suite 200 Seattle, WA 98104 plau...@cellnetix.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This email, 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 reply to the sender that you have received the message in error, then delete this message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Help! Losing sections from Superfrost Plus Slides
Prepare a 1% aq. sol. of Elmer's Glue and coat the slides with it. Oven dry and use the slides as usual. René J. --- On Wed, 12/9/09, Angelina Fong f...@zoology.ubc.ca wrote: From: Angelina Fong f...@zoology.ubc.ca Subject: [Histonet] Help! Losing sections from Superfrost Plus Slides To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Date: Wednesday, December 9, 2009, 2:40 PM Hi all, We have suddenly started losing tissue sections from our Superfrost Plus Slides. Our students have been cutting fixed, frozen, cryosections (20um) and thaw-mounting these onto Superfrost Plus slides. We have suddenly started losing lots of sections from these slides (again!!). The tissue is small - ie cross sections of frog aorta and longitudinal sections of nerves, so any lose of adhesion results in total loss of the tissue. The odd thing is that she is not losing every section on every slide, but half to 3/4 of the sections are falling off within the first rinse. The sections are from the same tissue block on the same slide while some falls and others don't. We are at a lose as to what we can do to rescue these sections. Does anyone know if there is any way to coat the slides in some solution with the tissue on them to help improve the adhesion without losing the ability to do immunofluorescence? Any further advice on cutting / drying protocols are welcomed. This keeps happening and the inconsistency of it has us so frustrated with this that we are thinking of going back to subbing our own slides. Thanks for your help! Angelina -- ~~o~~o~~o~~o~~o~~o~~o~~o~~o~~o Angelina Y. Fong, Ph.D. Department of Zoology Biological Sciences Building 6270 University Boulevard University of British Columbia Vancouver, BC, V6T 1Z4 Canada Ph: (604) 822-5799 Fax: (604) 822-2416 Email: f...@zoology.ubc.ca ___ 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] Help! Losing sections from Superfrost Plus Slides
I would suggest you check the lot#. You may have received a bad lot and need to get them replaced by the vendor. Hi all, We have suddenly started losing tissue sections from our Superfrost Plus Slides. Our students have been cutting fixed, frozen, cryosections (20um) and thaw-mounting these onto Superfrost Plus slides. We have suddenly started losing lots of sections from these slides (again!!). The tissue is small - ie cross sections of frog aorta and longitudinal sections of nerves, so any lose of adhesion results in total loss of the tissue. The odd thing is that she is not losing every section on every slide, but half to 3/4 of the sections are falling off within the first rinse. The sections are from the same tissue block on the same slide while some falls and others don't. We are at a lose as to what we can do to rescue these sections. Does anyone know if there is any way to coat the slides in some solution with the tissue on them to help improve the adhesion without losing the ability to do immunofluorescence? Any further advice on cutting / drying protocols are welcomed. This keeps happening and the inconsistency of it has us so frustrated with this that we are thinking of going back to subbing our own slides. Thanks for your help! Angelina -- ~~o~~o~~o~~o~~o~~o~~o~~o~~o~~o Angelina Y. Fong, Ph.D. Department of Zoology Biological Sciences Building 6270 University Boulevard University of British Columbia Vancouver, BC, V6T 1Z4 Canada Ph: (604) 822-5799 Fax: (604) 822-2416 Email: f...@zoology.ubc.ca ___ 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] maximum block cutting
Hi Histonet, I know we have discussed the average blocks histotechs cut per hour, but does anyone know if there is a maximum blocks that can be cut per hour? Or per day? We have regulations for cytotechs who are reading slides for maximum slides they can read per hour and are wondering if there is any similar regulation for histotechs. Thank you all for you help, Katelin Katelin Lester, HTL (ASCP) MedSurg Pathology Associates, Inc. (503) 443-2157 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: losing prefixed frozen sections from Plus charge slides
You wrote: We have suddenly started losing tissue sections from our Superfrost Plus Slides. Our students have been cutting fixed, frozen, cryosections (20um) and thaw-mounting these onto Superfrost Plus slides. We have suddenly started losing lots of sections from these slides (again!!). The tissue is small - ie cross sections of frog aorta and longitudinal sections of nerves, so any lose of adhesion results in total loss of the tissue. The odd thing is that she is not losing every section on every slide, but half to 3/4 of the sections are falling off within the first rinse. The sections are from the same tissue block on the same slide while some falls and others don't. We are at a loss as to what we can do to rescue these sections. Does anyone know if there is any way to coat the slides in some solution with the tissue on them to help improve the adhesion without losing the ability to do immunofluorescence? Any further advice on cutting / drying protocols are welcomed. This keeps happening and the inconsistency of it has us so frustrated with this that we are thinking of going back to subbing our own slides. Suggestions: 1. It could be a bad lot of slides, however, fixed frozen sections are notorious for falling off slides, even Plus charge. You did not say what fixative you used either? 2. I did not see where you cryoprotect your fixed tissue prior to snap freezing, and this may help, not only to reduce large ice water crystal damage, but also makes sectioning less of a crunchy affair. 3. Dry sections after sectioning in front of a fan at RT, and dry longer. Do NOT store just cut frozens in the cryostat (you didn't say how you handled the section immediately after picking up on the Plus Charge slides. Begin drying at RT immediately after sectioning. 4. Do not touch the surface of slides with fingers, sometime oils from skin transfer to slide surface. 5. It is a waste to money to coat expensive Plus Charge slides and coatings ruin or negate the Plus charge. If you carefully read the slide box insert, you will see this. Do not coat Plus charge slides with anything. If you want to coat slides with a gelatin (protein) subbing solution or Elmers glue (derived from milk products), use regular, clean microscope slides,, coat, air dry and store slides. . 6. Not knowing what or how you rinse, if the rinse is harsh, too fast, it will wash fixed sections off a slide. 7. 20 um is thick, although people do have success with careful and longer drying, depending on the staining method you want to perform. How you thaw this thick section onto a slide may be important. A young man taught us a clever way to do that by mounting section on a cold slide (cryostat chamber temperature, then thawing it an angle for a more gradual melting to surface. (don't put your finger under slide below section so it melts all at once onto the surface). He started at one edge of the section so it thawed from one side to another. He had FLAT sections and no bubbles under a thick fixed section. If your get any unevenness, the section may come up. Picking up a thick section from the blade holder plate may be part of the problem so try his method just for fun and possible success. Dry section well! 8. Fixing the tissue before doing frozen sections compromises the proteins, and once cross linked, tend to not like to stay on Plus charge slides, a common complaint seen on Histonet. Hence, subbing your own slides might be the answer if you had success before. 9. Make sure your blade is sharp so you obtain the flattest section possible. When sectioning, and you did not elaborate - mount first section on slide, lay slide outside cryostat, then pick up the next section next to the first section - we generally put first section closest to label, when picking up, and work down to bottom (away from label) for the rest of the sections if you are performing the mounting from the blade holder plate. Good luck, and be sure to check Histonet Archives for more answers. Gayle M. Callis HTL/HT/MT(ASCP) Bozeman MT ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] arizona state license
Does anyone know if Arizona requires a state license for histotechnicians/technologists? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] arizona state license
No we dont. At least not yet. However certification is strongly preferred around here. -Original Message- Date: Wednesday, December 09, 2009 4:39:04 pm To: 'histonet' Histonet@lists.utsouthwestern.edu From: dcoj...@tampabay.rr.com Subject: [Histonet] arizona state license Does anyone know if Arizona requires a state license for histotechnicians/technologists? ___ 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] Pathology Opportunity
Genitourinary / Urogenital Pathology Opportunity (Relocation Assistance Available) Location: Dallas Texas Fulltime / Permanent Position Generous relocation assistance available! Our client is one of healthcare’s leading clinical diagnostics organizations. Grow your career, as they expand their uropath practice. Your diagnostic expertise and professionalism will be rewarded in this position. Whether you are an established Genitourinary / Urogenital pathologist, or currently in a fellowship, you owe it to yourself to explore this opportunity. Keywords: Genitourinary, GU Pathology, Urogenital, Uropath Overview: Provide diagnostics expertise and clinical analysis on specimens. Specifically, this individual will be responsible for diagnosing reproductive and urinary system disorders and diseases utilizing clinical pathological diagnosis. Requirements: Significant experience in Genitourinary / Urogenital pathology is required, and leadership experience is preferred. Knowledge of diagnostic equipment and software applications. Medical Doctor or equivalent medical professional degree Board Certified or eligible in AP or AP/CP by the American Board of Pathology Fellowship training in Genitourinary / Urogenital pathology. Hold or eligible to obtain unrestricted medical licensure in Texas, Arizona, Massachusetts, and other states Strong scientific skills in surgical pathology Effective at collaboration, documentation presentation of findings, both written and oral. Duties Responsibilities: Conduct routine pathologic examination of biopsies Evaluate Genitourinary / Urogenital pathology specimens and render insightful and comprehensive diagnoses to enable clinicians to best treat their patients Consult with peer Pathologists on cases as requested or needed Participate in Quality Assurance programs, review cases, and participate in intradepartmental conferences to insure and improve accuracy of diagnoses and quality of care May be involved in research activities Reply To: Thomas Patrick Chuna, patricki...@sbcglobal.net Recruiter specializing in Biotech Professionals Thomas Patrick Chuna - Owner Patrick International Specialist In Scientific Information Management Opportunities Email: patricki...@sbcglobal.net Website: http://www.patrickinternational.net LinkedIn: http://www.linkedin.com/in/patrickinternational ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] maximum block cutting
There are no industry or regulatory standards that dictate a production maximum for blocks cut or slides produced hourly or daily. It is impossible to compare labs and set productivity standards becauseof the lack of standardized processes and protocols in Histology. I suggest that techs participating in a critical task, such as microtomy, for more than two hours consecutively, cannot consistently perform that task at an optimal productivity rate and error free. We should concenttrate our efforts on standardization of process and error reduction before considering setting production limits. William DeSalvo, B.S., HTL(ASCP) Date: Wed, 9 Dec 2009 14:07:37 -0800 From: histol...@medsurgpath.com To: histonet@lists.utsouthwestern.edu Subject: [Histonet] maximum block cutting Hi Histonet, I know we have discussed the average blocks histotechs cut per hour, but does anyone know if there is a maximum blocks that can be cut per hour? Or per day? We have regulations for cytotechs who are reading slides for maximum slides they can read per hour and are wondering if there is any similar regulation for histotechs. Thank you all for you help, Katelin Katelin Lester, HTL (ASCP) MedSurg Pathology Associates, Inc. (503) 443-2157 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _ Windows Live Hotmail gives you a free,exclusive gift. http://www.microsoft.com/windows/windowslive/hotmail_bl1/hotmail_bl1.aspx?ocid=PID23879::T:WLMTAGL:ON:WL:en-ww:WM_IMHM_7:092009___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Help! Losing sections from Superfrost Plus Slides
Hi Angelina, We were having the same problem with frozen mouse aortas. I was cutting at 5ums onto superfrost slides and was losing about half of my sections. I ended up swapping to superfrost plus ultra, plus I bake the slides in the histology oven at 60 degress celcius for 10 minutes and now I nearly never lose a section - the only time I do is when I have done a lazy job of cutting and have collected a section with a fold in it. Good luck! Message: 9 Date: Wed, 09 Dec 2009 11:40:50 -0800 From: Angelina Fong f...@zoology.ubc.ca Subject: [Histonet] Help! Losing sections from Superfrost Plus Slides To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Message-ID: 4b1ffd42.2040...@zoology.ubc.ca Content-Type: text/plain; charset=ISO-8859-1; format=flowed Hi all, We have suddenly started losing tissue sections from our Superfrost Plus Slides. Our students have been cutting fixed, frozen, cryosections (20um) and thaw-mounting these onto Superfrost Plus slides. We have suddenly started losing lots of sections from these slides (again!!). The tissue is small - ie cross sections of frog aorta and longitudinal sections of nerves, so any lose of adhesion results in total loss of the tissue. The odd thing is that she is not losing every section on every slide, but half to 3/4 of the sections are falling off within the first rinse. The sections are from the same tissue block on the same slide while some falls and others don't. We are at a lose as to what we can do to rescue these sections. Does anyone know if there is any way to coat the slides in some solution with the tissue on them to help improve the adhesion without losing the ability to do immunofluorescence? Any further advice on cutting / drying protocols are welcomed. This keeps happening and the inconsistency of it has us so frustrated with this that we are thinking of going back to subbing our own slides. Thanks for your help! Angelina -- ~~o~~o~~o~~o~~o~~o~~o~~o~~o~~o Angelina Y. Fong, Ph.D. Department of Zoology Biological Sciences Building 6270 University Boulevard University of British Columbia Vancouver, BC, V6T 1Z4 Canada Ph: (604) 822-5799 Fax: (604) 822-2416 Email: f...@zoology.ubc.ca -- Message: 10 Date: Wed, 9 Dec 2009 15:08:57 -0500 From: HistoLab histosea...@gmail.com Subject: [Histonet] Help! Losing sections from Superfrost Plus Slides To: histonet@lists.utsouthwestern.edu Message-ID: 521c6d260912091208n421bf467g3240121cd26f0...@mail.gmail.com Content-Type: text/plain; charset=ISO-8859-1 Angelina, I have seen this post a few times before and sometimes the problem was the drying technique or making sure your water bath is clean and free of any debris. I also remember hearing about someone pre-treating their slides with a Trilogy (EDTA) buffer in a pressure cooker. Good Luck! *Matthew Semovoski* Sales Manager Gorilla Scientific Corporation 1-866-435-4977 www.gorillascientific.com _ Get more out of Hotmail Check out the latest features today http://windowslive.ninemsn.com.au/hotmail/article/878466/your-hotmail-is-about-to-get-even-better___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Leitz 1512
Several of you have recently written wanting the instructions for the Leitz 1512. I had temporarily misplaced these because my lab moved for the second time in the past year. I have now located these. Please re-connect with me so I can fax or mail you these instructions. Thank you. Sharon Osborn ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
