[Histonet] Thermo Stainmate Max
Hi, Have any of you used the Thermo Stainmate Mix? It has 10 dishes holding 200ml of fluid? Am looking for opinions. We are considering getting this for our rush FNAs. Thanks. __ Lynn O'Donnell, CT (ASCP), MHA Technical Specialist, Cytology Danbury Hospital 203-739-7846 lynn.o'donn...@wchn.org ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: formalin substitutes. HELP
Hello Nieves, Here in the US Anatech LTD sells a gyloxal fixative called Prefer. I have attached a link to the website. Go to the MSDS menu and click on 'Prefer' to pull up the MSDS. http://www.anatechltdusa.com/ Below is an old post from the HistoNet that might be helpful. I suggest you contact Ada Feldman at Anatech, as she should be able to address your issues Regards, Brett Brett M. Connolly, Ph.D. Principal Scientist, Imaging Dept. Merck Co., Inc. PO Box 4, WP-44K West Point, PA 19486 brett_conno...@merck.com T- 215-652-2501 F- 215-993-6803 As the developers of the Prefer fixative we would like to address some of the issues. Interchangeability of glyoxal products: The manufacturer of each fixative should be able to provide the information necessary to work with their fixative in conjunction with any other fixative that a lab may be using. Just as an example, Hollandes is not compatible with NBF and requires special attention. As for Prefer we can say it is compatible with the majority of other fixatives, glyoxal or not. Limited time in the fixative: There is a slight reduction in staining intensity after several weeks, but increasing staining time corrects this. So tissues are not rendered unstainable. Transition period: This is true any time you are switching fixatives or processing methods. Eosinophila: Your statements here are true. Lysis of erythrocytes: True again. It is often seen as an advantage because diagnostics cells are easier to see. Breast cancer: It would seem that the improved nuclear morphology would make marked nuclear variation easier to determine. Prostate biopsies: Hope you can get a response from any of the glyoxal users with prostate biopsies. We would be interested in this answer also. Ada T. Feldman, MS, HT/HTL(ASCP) ANATECH LTD. 1020 Harts Lake Road Battle Creek, MI 49015 Phone: 800.262.8324 Fax: 269.964.8084 email: adafeld...@anatechltdusa.com website: www.anatechltdusa.com On Jul 14, 2006, at 4:13 PM, rsrichm...@aol.com wrote: The people at Anatech, makers of Prefer fixative, have published a review of glyoxal fixation that every pathologist and histotechnologist ought to read. This working surgical pathologist would like to add - and solicit - some comments on Histonet. Glyoxal Fixation and Its Relationship to Immunohistochemistry. Richard W. Dapson, Ada T. Feldman, and Dee Wolfe. Anatech Limited, Battle Creek MI. The Journal of Histotechnology June 2006;29:65-76. I don't want to change, but I think we all need to be prepared for the day when a manager walks into our laboratory, or a letter from a regulatory agency arrives in the mail, telling us that we have to get rid of formaldehyde right now. Probably glyoxal is the only acceptable substitute, and we all need to have a look at it. I have a number of questions. Interchangeability of glyoxal products: Prefer is described as a buffered solution of glyoxal with a pH of about 4. The formula is a trade secret. Competing glyoxal products probably also have trade-secret formulas. So can a lab change brands of buffered glyoxal without problems, or does it have to stay with a particular brand with its trade-secret buffering? - We've seen a similar problem with distilling aliphatic xylene substitutes: every one of them requires a separate distillation routine, at least on a spinning-band still. - I'll leave it to John Kiernan to comment on the appropriateness of trade- secret reagents in histopathology. Limited time in the fixative: Tissue can be left in neutral buffered formalin for quite a long time and still be stainable, but tissue stored in glyoxal becomes unstainable after about two weeks. Can glyoxal fixed tissue be transferred to 70% ethanol for more prolonged storage? - A very occasional surgical specimen requires additional blocks after a week - a bigger problem will be the pathologist who doesn't trim his autopsies promptly. Transition period: A laboratory changing to glyoxal would have to keep IHC procedures for both fixatives working for some time. There would have to be some way to identify whether a block was fixed in formaldehyde or glyoxal. Eosinophilia: One ought to be able to distinguish eosinophils from neutrophils in tissue sections by nuclear morphology, without having to see granules. But quantitation of eosinophils - needed in an increasing number of GI biopsy situations - could be a problem. We might need an IHC for eosinophils in some of these settings. Lysis of erythrocytes: Not much of a problem, since we're used to it with acid fixatives anyway. Breast cancer: Elimination of nuclear bubble artifact in breast biopsy specimens may raise the apparent nuclear grade of tumors, and thus increase Nottingham (Elston-Ellis) scores. Prostate biopsies: I'd want to see some prostate biopsies - is somebody from OURLab in
Re: [Histonet] formalin substitutes. HELP
Nieves: NO formalin substitute will work in the same way as formalin and the solution is not to start testing other substitutes that will make your life miserable and your sections and staining procedures of sub-standard quality. The solution is: 1- to use LESS amounts of formalin (a 5:1 volume is more than enough); 2- have a well ventilated area; 3- do NOT prepare your own buffered formalin; buy pre-filled sample bottles; and 4- never handle formalin more than absolutely necessary. Under separate cover I am sending you 2 articles I wrote on this subject René J. From: Nieves Gomez ngo...@neiker.net To: histonet@lists.utsouthwestern.edu Sent: Friday, November 9, 2012 8:55 AM Subject: [Histonet] formalin substitutes. HELP Dear Histonetters, I'm new in the net. I work as Pathologist in a Vet Lab in Spain. Because formalin is toxic our Lab is for the practice of using alternative fixatives. I think the main viable alternative is glyoxal based formulas, but I have so many questions that Commercials don't know or don't want to answer me. For example, have a MSDS or is it accessible? Really is less hazardous than formalin or just is not checked? (the advantages and desadvantages of formalin are known for at least 100 years). Related to this, I think the glyoxal is suggested as a formalin substitute in an article in 1940's and now it is sold as a new product and most of the products are sold as green, no-toxic or non harmful. In my opinion, a fixative can not be non-toxic if you want it fixed tissues. Another question is the time needed to fix tissues or the ratio volume specimen/fixative. To the first point, I have read an article that mentions there is mould growth in specimens over time. Are we changing a chemical risk to a biological risk? In my lab we have a specifically workstation for the gross examination and sectioning of specimens, and we wear all the Personal Protective Equipment needed (formalin chemical filters, gloves, googles...) that minimizes the risk (the chemical risk not the biological risk). It is believed that formalin given time will kill any microorganisms (or spores) present in tissues, mycobacteria also.. what about these new products? Are they germicidal? I do not get to appreciate the morfological changes (nuclear changes, lysis of erithrocites, eosinophilia...) because they are well documented. My aim is to know if there is any Lab that works with any formalin substitute routinely to aks these questions. Please, help me. Thanks and have a nice weekend Nieves Gomez Veterinary Pathologist Animal Health Department NEIKER-Tecnalia Berreaga, 1. 48160 Derio. Bizkaia. Spain. ngo...@neiker.net ___ 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] Stain for osteoporotic bone in Spurr plastic
Allison, I had a great von kossa protocol for this but it was on glycol methacrylate embedded bone, here it is, it uses the fluorescence property of eosin which will stain the osteoid with the calcified bone covered by the silver von kossa. GMA is not removed Rinse 5 micron sections in dih20, use clean glassware Make 2% silver nitrate, put on the slides and expose them to uv (a lamp or even in the window will do, for about 20 min. The calcified bone should turn black Rinse well in dih20, you can fix the silver with sodium thiosulfate but do it quickly as it does remove some silver, rinse well in dih20 Hematoxylin nuclear stain, we used Gills#3 for 5 min., wash in tap water, blue in ammonia water, wash well in tap water then to dih20 Make a 0.2% solution of aqueous eosin y, stain slides in this for 20 min., rinse in dih20 and airdry, coverslip with permount. You can measure the calcified bone and nuclei in the light microscope, then use a fluorescent scope with a fitc filter to see the osteoid, it will be green just on the surface of the calcified bone which is black from the silver. We got a total sample area measurement, calcified area as a percentage of total area, and osteoid area as a percentage of total area. We did other measurements as well such as number of osteoclasts (we used a Acid phosphotase (TRAP) enzyme histochemical stain for this) and number of osteoblast (can use alkaline phosphotase enzyme histochemical stain for these or we just counted them from the VK slide with the heme nuclear stain by morphology, they will be lining the surface of the bone, very flat and run together.) Hope this helps. Goldner TC is also used to measure osteoid but this worked best for us. Patsy Ruegg, HT(ASCP)QIHC Ruegg IHC Consulting, LLC 40864 Arkansas Ave Bennett, CO 80102 Phone: 303-644-4538 Fax: 720-859-4110 pru...@ihctech.net -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jack Ratliff Sent: Thursday, November 08, 2012 4:52 PM To: Malandra, Allison E; Histonet Subject: RE: [Histonet] Stain for osteoporotic bone in Spurr plastic Allison, I am not exactly certain if this protocol will work for Spurr's embedded specimens, but here is the protocol that I use for deplasticized sections of methyl methacrylate embedded undemineralized bone (non-decalcified): Xylenes - warmed @ 50 C for 30 min w/ dip dunk @ 15 min interval Xylenes - warmed @ 50 C for 30 min w/ dip dunk @ 15 min interval Xylenes - warmed @ 50 C for 30 min w/ dip dunk @ 15 min interval 100% EtOH - room temp for 5 min 95% EtOH - room temp for 5 min 70% EtOH - room temp for 5 min DI H2O - room temp for 5 min Silver Nitrate (20 g) + DI H2O (400 ml) - room temp (in dark protected from light) for 5 min DI H2O rinse - room temp (protected from light) for 1 min DI H2O rinse - room temp (protected from light) for 1 min DI H2O rinse - room temp (protected from light) for 1 min Sodium Carbonate (22.5 G) + DI H2O (337.5 ml) + 37% Formaldehyde (112.5 ml) - room temp (protected from light) for 2 min DI H2O rinse - room temp for 1 min DI H2O rinse - room temp for 1 min Sodium Thiosulfate (40 g) + Potassium Ferricyanide (2 g) + DI H2O (420 ml) Solution - room temp for 30 sec(solution stable for 30-60 min after addition of potassium ferricyanide) Running Tap Water Rinse - 10 min Counterstain w/ MacNeal's Tetrachrome (12 g) + DI H2O (600 ml) - room temp for 6-8 min(stir continuously w/ heat @ 60C for 48 hours covered, then filter with paper towel) DI H2O rinse - room temp for 1 min DI H2O rinse - room temp for 1 min DI H2O rinse - room temp for 1 min 70% EtOH - room temp for 1 min 100% EtOH - room temp for 1 min Xylenes - room temp for 3 min Xylenes - room temp for 3 min Coverslip w/ Eukitt Hope this helps! Please feel free to contact me if you have any questions or concerns. Best Regards, Jack Jack L RatliffOwner, Ratliff Histology Consultants, LLCChairman, Hard Tissue Committee - National Society for Histotechnology (317) 281-1975jratl...@ratliffhistology.com LinkedIn Profile: http://www.linkedin.com/profile/edit?trk=hb_tab_pro_top From: allison-malan...@uiowa.edu To: histonet@lists.utsouthwestern.edu Date: Thu, 8 Nov 2012 22:52:42 + Subject: [Histonet] Stain for osteoporotic bone in Spurr plastic Hi there - We are trying to stain osteoporotic bone that was embedded using Spurr plastic. We need to be able to differentiate osteoid from mineralized bone. We have tried Gio's trichrome and Goldner's trichrome with no success, both surface staining and on deplasticized slides. We are thinking of trying Von Kossa, does anyone have a recipe/protocol for this? Or other types of stains that you have had success with? Thank you! Allison Malandra, DVM University of Iowa, Bone Healing Research Lab Notice: This UI Health Care e-mail (including attachments) is covered by the Electronic Communications Privacy
[Histonet] decal affecting IHC
Does decalcifying tissue (Calex II) affect the antibody reaction for IHC in bony tissue? Diana ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] decal affecting IHC
I do not know what is in Calex II but high concentration HCl can have a negative affect. We use 5% formic acid without issue. Jack Jack L Ratliff Owner/Histologist, Ratliff Histology Consultants, LLC Chairman, Hard Tissue Committee - National Society for Histotechnology On Nov 9, 2012, at 11:34 AM, Diana McCaig dmcc...@ckha.on.ca wrote: Does decalcifying tissue (Calex II) affect the antibody reaction for IHC in bony tissue? Diana ___ 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] decal affecting IHC
IHC reactions can be affected by decalcification especially if: 1- the tissue is not perfectly fixed before going into decalcification 2- decalcification is done at temperature above room temp. 3- decal solution is too acid, specially made with inorganic acids. If at all possible decalcification should be done with a chelating agent, like EDTA René J. From: Diana McCaig dmcc...@ckha.on.ca To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu Sent: Friday, November 9, 2012 12:34 PM Subject: [Histonet] decal affecting IHC Does decalcifying tissue (Calex II) affect the antibody reaction for IHC in bony tissue? Diana ___ 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] decal affecting IHC
Diana I agree with Jack, formic acid is the way to go in my opinion, we use 10% formic acid routinely but have pushed it up to 20% on occasion when time was an issue. HCL can work if its controlled really well but you don't have much wiggle room, you can over decal quite quickly, formic acid is a bit more forgiving. As for Rene's comment we have seen the exact opposite with respects to the EDTA decal part. Granted we do not run a lot of IHC on EDTA decaled samples but on several occasions and with several antibodies we were able to obtain good IHC staining in formic acid decaled samples but those antibodies did not work well in EDTA decaled samples. These were not direct comparisons of the decalcification method on the same study, but on samples from different studies so other factors could have affected the results. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, CO 80308-1592 (303) 682-3949 office (303) 682-9060 fax (303) 881-0763 cell www.premierlab.com Ship to address: 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 Rene J Buesa Sent: Friday, November 09, 2012 10:47 AM To: Diana McCaig; 'histonet@lists.utsouthwestern.edu' Subject: Re: [Histonet] decal affecting IHC IHC reactions can be affected by decalcification especially if: 1- the tissue is not perfectly fixed before going into decalcification 2- decalcification is done at temperature above room temp. 3- decal solution is too acid, specially made with inorganic acids. If at all possible decalcification should be done with a chelating agent, like EDTA René J. From: Diana McCaig dmcc...@ckha.on.ca To: 'histonet@lists.utsouthwestern.edu' histonet@lists.utsouthwestern.edu Sent: Friday, November 9, 2012 12:34 PM Subject: [Histonet] decal affecting IHC Does decalcifying tissue (Calex II) affect the antibody reaction for IHC in bony tissue? Diana ___ 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] Becky Orr
I'm looking to contact Rebecca (Becky) Orr. Does anyone know her current email address? The one I have bounced back. Thanks, Andrea ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Microtome knives
Greetings I need some advice regarding microtome knives. I am not histo tech, I did all my sectioning in a plant research lab, but now I find myself needing to learn more about histo type methods. We have microtomes, AO 820's, and we have a bunch of donated knives. I need advice about whether it would be better to find a knife sharpener and use the microtome knives we have, or check into getting a disposable knife holder. When I was sectioning, we just used a simple razor blade holder. Now I see references to high profile and low profile blades and holders, and I don't know the difference. Anyone willing to help me out? Thanks Jon Jonathan Krupp Delta College 5151 Pacific Ave. Box 212 Stockton, CA 95207 209-954-5284 jkr...@deltacollege.edu Find us on Facebook @ Electron Microscopy at SJ Delta College ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Microtome knives
Because of your donated knives you will have to buy a knives sharpener which are costly and not very easy to find. Your best option is to buy a high profile disposable blades holder (that will be cheaper), buy disposable blades and avoid all the frustrations and waste of time sharpening knives. René J. From: Jon Krupp jkr...@deltacollege.edu To: Histonet@lists.utsouthwestern.edu Sent: Friday, November 9, 2012 1:49 PM Subject: [Histonet] Microtome knives Greetings I need some advice regarding microtome knives. I am not histo tech, I did all my sectioning in a plant research lab, but now I find myself needing to learn more about histo type methods. We have microtomes, AO 820's, and we have a bunch of donated knives. I need advice about whether it would be better to find a knife sharpener and use the microtome knives we have, or check into getting a disposable knife holder. When I was sectioning, we just used a simple razor blade holder. Now I see references to high profile and low profile blades and holders, and I don't know the difference. Anyone willing to help me out? Thanks Jon Jonathan Krupp Delta College 5151 Pacific Ave. Box 212 Stockton, CA 95207 209-954-5284 jkr...@deltacollege.edu Find us on Facebook @ Electron Microscopy at SJ Delta College ___ 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] Microtome knives
Hi Jon, Depends on your cash flow. You could get a used sharpener somewhere off the web somewhere such as http://www.labx.com/v2/adsearch/resultsnew.cfm?sw=sharpenermr=25te=cat , or http://www.medwow.com/used-microtome-knife-sharpener-equipment/63.med but sharpening knives is a pain IMO and steel knives present more of a safety hazard. I would recommend a sharpener that uses the glass honing plates. You would also need the coarse and fine abrasives. Personally, I would opt for a low profile disposable blade holder that fits your 820. Low and high profile refer to the size (height) of the blade. We use low profile for paraffin block sectioning and high profile for cryostat sectioning. Brett Brett M. Connolly, Ph.D. Principal Scientist, Imaging Dept. Merck Co., Inc. PO Box 4, WP-44K West Point, PA 19486 brett_conno...@merck.com T- 215-652-2501 F- 215-993-6803 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Jon Krupp Sent: Friday, November 09, 2012 1:50 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Microtome knives Greetings I need some advice regarding microtome knives. I am not histo tech, I did all my sectioning in a plant research lab, but now I find myself needing to learn more about histo type methods. We have microtomes, AO 820's, and we have a bunch of donated knives. I need advice about whether it would be better to find a knife sharpener and use the microtome knives we have, or check into getting a disposable knife holder. When I was sectioning, we just used a simple razor blade holder. Now I see references to high profile and low profile blades and holders, and I don't know the difference. Anyone willing to help me out? Thanks Jon Jonathan Krupp Delta College 5151 Pacific Ave. Box 212 Stockton, CA 95207 209-954-5284 jkr...@deltacollege.edu Find us on Facebook @ Electron Microscopy at SJ Delta College ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Notice: This e-mail message, together with any attachments, contains information of Merck Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates Direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Dermatology KOH testing
Does anyone perform KOH tests? If so, could you please email me and give me information on your process. Thanks, Trini ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] test
Just testing-new subscriber ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Diane Tokugawa/CA/KAIPERM is out of the office.
I will be out of the office starting 11/09/2012 and will not return until 11/15/2012. Note: For Cytology issues, please call Robin (day) at 8-421-5040, Wanda (day) 8-421-5426, or Eric (swing) 8-421-5405 For Histology issues, please call Mario (day) 8-421-4961, Barbara (swing) 8-421-4959, Bob (IHC) 8-421-4706, Kiran at 8-421-5404, or general histology client service at 8-421-5408 or Lotus Notes.___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Mold Release Problems
Does anyone know of a reference for the problems from using mold release? When too much is used on the mold it eats into the block and causes the ribbons to explode on the water bath and creates an artifact in the tissue.I cannot find a reference for it or any published material about the problem but have been to seminars where it was discussed.When it is not used the problem never occurs. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
