[Histonet] Paraffin Blocks
Hello Histoneters, Maybe one of you could assist me in search of some cancer blocks. I am in search for Ewing's sarcoma, rhabdomyosarcoma, Mesothelioma, and Small Cell Sarcoma. I am looking for true patient disease tissue not a clone. If anyone has any additional block or fixed tissue they could spare it would be very much appreciated. We will purchase these blocks from you or your institution. Please contact me if you can assist me. Thank you. Regards, Clarence Owens, HT (ASCP) QIHC Sent from my iPhone ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC on fish
Cate - Your gut is probably right but results are unpredictable. I have forwarded your query to the Bozeman Fish Technology center to see if they can offer any assistance. Tresa -Original Message- From: Hardy, Cate [mailto:cha...@csu.edu.au] Sent: Sunday, June 14, 2015 4:10 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC on fish Hi all; Has anyone performed IHC on fish. I have been asked to perform this using several of our routine antibodies that I use on mammals. My gut tells me it won't work but I am a lowly histotech and don't know anything. Seeking knowledge from learned tech's with experience in such matters Thanks Cate Hardy Senior Technical Officer Veterinary Diagnostic Laboratory Charles Strut University NSW Australia Charles Sturt University | ALBURY-WODONGA | BATHURST | CANBERRA | DUBBO | GOULBURN | MELBOURNE | ONTARIO | ORANGE | PORT MACQUARIE | SYDNEY | WAGGA WAGGA | LEGAL NOTICE This email (and any attachment) is confidential and is intended for the use of the addressee(s) only. If you are not the intended recipient of this email, you must not copy, distribute, take any action in reliance on it or disclose it to anyone. Any confidentiality is not waived or lost by reason of mistaken delivery. Email should be checked for viruses and defects before opening. Charles Sturt University (CSU) does not accept liability for viruses or any consequence which arise as a result of this email transmission. Email communications with CSU may be subject to automated email filtering, which could result in the delay or deletion of a legitimate email before it is read at CSU. The views expressed in this email are not necessarily those of CSU. Charles Sturt University in Australia http://www.csu.edu.au The Grange Chancellery, Panorama Avenue, Bathurst NSW Australia 2795 (ABN: 83 878 708 551; CRICOS Provider Numbers: 5F (NSW), 01947G (VIC), 02960B (ACT)). TEQSA Provider Number: PV12018 Charles Sturt University in Ontario http://www.charlessturt.ca 860 Harrington Court, Burlington Ontario Canada L7N 3N4 Registration: www.peqab.ca Consider the environment before printing this 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
Re: [Histonet] IHC on fish
I stained some zebra fish many many years ago. For the life of me, I can't remember with what! It's all going to come down to whether or not your Abs can recognize it. I'd say give it a go and don't trust a negative result or look for Abs that should specifically work for fish. Not all companies even test that obviously but there are a few that do. I used to work for a company that would test Abs for western blots against pretty much anything that would go in a blender! :) What antigens are you looking for? Good luck. Katy Message: 1 Date: Sun, 14 Jun 2015 20:09:40 +1000 From: Hardy, Cate cha...@csu.edu.au To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC on fish Message-ID: 144c8e2433ca3b4d9e080e5cda803d05a752cc2...@mail01.csumain.csu.edu.au Content-Type: text/plain; charset=us-ascii Hi all; Has anyone performed IHC on fish. I have been asked to perform this using several of our routine antibodies that I use on mammals. My gut tells me it won't work but I am a lowly histotech and don't know anything. Seeking knowledge from learned tech's with experience in such matters Thanks Cate Hardy Senior Technical Officer Veterinary Diagnostic Laboratory Charles Strut University NSW Australia Charles Sturt University | ALBURY-WODONGA | BATHURST | CANBERRA | DUBBO | GOULBURN | MELBOURNE | | ONTARIO | ORANGE | PORT MACQUARIE | SYDNEY | WAGGA WAGGA | ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] IHC on fish
Cate - Why not do a Blast search for the sequences your anti-mammal antibodies recognize see if they occur in fish? Some proteins are strongly conserved, or have conserved domains - you might get lucky! I tried a new (to me) antibody last week that was made against an amoeba, and it worked gangbusters on mouse and rat. Tamara ... Tamara Howard Dept. of Cell Biology Physiology University of New Mexico Albuquerque, NM Message: 1 Date: Sun, 14 Jun 2015 20:09:40 +1000 From: Hardy, Cate cha...@csu.edu.au To: histonet@lists.utsouthwestern.edu histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC on fish Message-ID: 144c8e2433ca3b4d9e080e5cda803d05a752cc2...@mail01.csumain.csu.edu.au Content-Type: text/plain; charset=us-ascii Hi all; Has anyone performed IHC on fish. I have been asked to perform this using several of our routine antibodies that I use on mammals. My gut tells me it won't work but I am a lowly histotech and don't know anything. Seeking knowledge from learned tech's with experience in such matters Thanks Cate Hardy Senior Technical Officer Veterinary Diagnostic Laboratory Charles Strut University NSW Australia Charles Sturt University | ALBURY-WODONGA | BATHURST | CANBERRA | DUBBO | GOULBURN | MELBOURNE | ONTARIO | ORANGE | PORT MACQUARIE | SYDNEY | WAGGA WAGGA | LEGAL NOTICE This email (and any attachment) is confidential and is intended for the use of the addressee(s) only. If you are not the intended recipient of this email, you must not copy, distribute, take any action in reliance on it or disclose it to anyone. Any confidentiality is not waived or lost by reason of mistaken delivery. Email should be checked for viruses and defects before opening. Charles Sturt University (CSU) does not accept liability for viruses or any consequence which arise as a result of this email transmission. Email communications with CSU may be subject to automated email filtering, which could result in the delay or deletion of a legitimate email before it is read at CSU. The views expressed in this email are not necessarily those of CSU. Charles Sturt University in Australia http://www.csu.edu.au The Grange Chancellery, Panorama Avenue, Bathurst NSW Australia 2795 (ABN: 83 878 708 551; CRICOS Provider Numbers: 5F (NSW), 01947G (VIC), 02960B (ACT)). TEQSA Provider Number: PV12018 Charles Sturt University in Ontario http://www.charlessturt.ca 860 Harrington Court, Burlington Ontario Canada L7N 3N4 Registration: www.peqab.ca Consider the environment before printing this email. -- ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] ammonium bromide in fixative
Thank you for such a detailed response. It is doubtful I would have found this information without you. The National Histology Society online learning center is one of my study resources, this is where the question came from. They also include several mercuric fixatives that we would never use in our lab in this day and age. Perhaps the material has not been updated for a number of years. I have not yet heard from any recent exam takers about the content of old techniques. I find them interesting, but don't know how much time to devote to them. Katherine S Walters Histology Director Central Microscopy Research Facility University of Iowa 76 Eckstein Medical Research Building 431 Newton Road Iowa City, Iowa 52242 319-335-8142 Facility Website: http://cmrf.research.uiowa.edu/ From: John Kiernan [mailto:jkier...@uwo.ca] Sent: Friday, June 12, 2015 11:58 PM To: Walters, Katherine S; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] ammonium bromide in fixative Formalin with ammonium bromide is the fixative prescribed for traditional silver and gold techniques for staining neuroglial/x cells in free-floating frozen sections. Adding ammonium bromide lowers the pH of the fixative because of a chemical reaction with formaldehyde. An Argentinian histologist/x called Lascano showed in the 1940s that any sufficiently acidified formaldehyde solution (pH 1.5) was just as good as FAB for Cajal's gold-sublimate (astrocytes) and for silver carbonate methods (oligodendrocytes, microglia). The reaction that lowers the pH is 4NH4+ + 6HCHO --- C6H12N4 + 6H2O + 4H+ (Subscripts and superscripts in this equation will not be honoured in this email!) The 4H+ lowers the pH. The other product, C6H12N4 is hexamethylenetetramine, also known as hexamine in Britain and methenamine in the USA, and used in various histochemical staining methods, notably Grocott's method for fungal cell-walls in sections of animal (including human) tissues. The bromide (Br-) ion of NH4Br is irrelevant, in the FAB fixative and also in Globus's pretreatments (dilute ammonia followed by dilute hydrobromic acid). Globus's bromuration treatment was applied to frozen sections of pieces of CNS that had been fixed in non-acidified formalin. The references for Lascano's work are: Lascano, E.F. (1946a). Influencia del pH en la impregnacion argentica del tejido nervioso. Archivos de la Sociedad Argentina de Anatomia Normal y Patologica 8:105-114. Lascano, E.F. (1946b). Importancia del pH en la fijacion del tejido nervioso. Creacion artificial de fijadores/x tipo formol-bromuro y formol-nitrato de urano de Cajal. Archivos de la Sociedad Argentina de Anatomia Normal y Patologica 8:185-194. Lascano, E.F. (1946c). Influencia del pH del fijador en la coloracion argentica del tejido nervioso. Archivos de la Sociedad Argentina de Anatomia Normal y Patologica 8:272-276. Not everyone agreed with Lascano! Polak, M. (1948). Sobre la importancia del bromuro de amonio de la solucion fijadora de Cajal en la impregnacion argentica del tejido nervioso. Archivos de la Sociedad Argentina de Anatomia Normal y Patologica 10:224-234. Do you really need this information to pass your HTL qualifying exam? It isn't easily found in books or with Google Scholar. I came across these papers quite by chance when looking over some old journals that UWO's library had set aside for throwing out (Gasp!), about 1980. Yes, they had hit a new low! Their services have, however, been exceptionally good for the last 10-15 years. Does anyone still use either Cajal's method for astrocytes or traditional silver carbonate methods to stain oligodendrocytes and microglia? They are difficult for several reasons, take up time, and require an old-fashioned freezing microtome to cut and collect the rather thick sections that are needed. Reliable antibodies for immunostaining glial cell-types have been available for many years, and they work on any kind of section. You need some thickness to appreciate the 3D shapes of astrocytes and oligodendrocytes, however they are stained. There's plenty of histo-history in the traditional neuroglia stains, which defined the cell-types for identification by electron microscopy and immunohistochemistry. John Kiernan Anatomy Cell Biology University of Western Ontario London, Canada = = = On 11/06/15, Walters, Katherine S katherine-walt...@uiowa.edumailto:katherine-walt...@uiowa.edu wrote: Hi all, I am studying to take the HTL certification test and ran across a reference to Formalin Ammonium Bromide. I see that it is very good for central nervous tissue fixation, it must be made fresh and that its pH is 1.5. Does anyone happen to know the reason for ammonium bromide in this fixative? I have been looking online and this has not been explained. Also, has anyone taken this test lately? I am curious as to how much old techniques, such as mercuric fixatives will be included?
Re: [Histonet] IHC on fish
Hello, From: Hardy, Cate cha...@csu.edu.au Subject: [Histonet] IHC on fish Hi all; Has anyone performed IHC on fish. I have been asked to perform this using several of our routine antibodies that I use on mammals. My gut tells me it won't work but I am a lowly histotech and don't know anything. Seeking knowledge from learned tech's with experience in such matters Thanks My entire thesis was about taking antibodies generated against mammalian antigens and using them to stain equivalent structures in fish. In many cases the results were as expected. I didn't modify the protocols in any way. Simply use the same protocol you are using for the mammals and apply them to the fish. What did save me a lot of time was doing western blots first to test for immunoreactivity and specificity. Some of my pretty micrographs had to be discarded because on a western blot the antibody labeled multiple bands irregardless of the antibody dilution. If you know the immunogen sequences blast the various fish protein sequence databases and see how well the antigen is conserved in various species. If there is low similarity in the sequence then it isn't worth testing the antibody. Again, confirm with a western blot or dot-blot for immunoreactivity. As regards fish sectioning the little bones can be a bother, and the tissues seems to dry out and harden very fast in ethanol. I would suggest something like Davidson's fixative, overnight at 4 oC and then quickly process the tissue. For antigen retrieval you can modify the protocol of http://www.ncbi.nlm.nih.gov/pubmed/21603650 for sections. You can also try Citrate Buffer Antigen retrieval. If you are going to do immunofluorescence then the protocol I got from Gayle Callis works well: after taking water incubate slides or section twice in 300 mM glycine in TBS, pH 7.4, 15 minutes at room temperature (RT) to quench autofluorescence. Good luck with the antibodies! -- Tyrone Genade Orange City, Iowa tel: (+1) 712 230 4101 http://tgenade.freeshell.org Romans 6:23: The gift of God is eternal life through Christ Jesus our Lord. To find out how to receive this FREE gift visit http://www.alpha.org. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Processing Validation
Does anyone have a processing validation procedure they would like to share? Thank you! Mary Leslie ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] FT position HISTOTECHNOLOGIST/IHC Delray Beach, FL
Hi Histonetters! We are looking for a full time licensed histotech here in our very busy Delray Florida Dermatology Lab. This is a permanent full time SECOND SHIFT (40 hours) position with benefits (medical/401k/vacation) and competitive pay. THIS IS A DRUG FREE WORKPLACE. Drug testing, background check and personality testing is required. ONLY SERIOUS INQURIES, please read EVERY qualification desired. Sorry, NO relocation assistance offered. Position available for training immediately. ***PLEASE NO HEAD HUNTERS/PLACEMENT SERVICES***!!! Email your resume to lengim...@leavittmgt.commailto:lengim...@leavittmgt.com if interested. *Full time position Mon-Fri 2p-10:30p (IMMUNOHISTOCHEMISTRY/SPECIALS department) *MUST be licensed as a FLORIDA HISTOTECHNOLOGIST (this is NOT negotiable) *EXPERIENCE WITH IHC A MUST! Leica (BOND) and Roche/Ventana equipment experience preferred *Must be able to multi-task special stains *MUST have at LEAST 2 years experience. Please DO NOT respond if you do not possess EXPERIENCE in this area! *must be confidant, quick learner, self motivated, reliable and a team player Kari M Simeone Histology/Immunohistochemistry Specialist Supervisor The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If you have received this message in error, you are prohibited from copying, distributing or using the information. Please contact the sender immediately by return e-mail and delete the original message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Processing Validation
Would love to see also. Michael Ann On 6/15/15, 2:18 PM, Leslie, Mary mary.les...@tricore.org wrote: Does anyone have a processing validation procedure they would like to share? Thank you! Mary Leslie ___ 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
