Excellent! Happy to read the venerable Neil Hand is coming back to the symposium, always a great speaker!
-Damien On Tue, Mar 13, 2012 at 10:50 AM, Jack Ratliff <ratliffj...@hotmail.com>wrote: > > I might also add that Neil Hand is co-speaking with myself and Philip > Seifert this year at the annual National Society for Histotechnology - > Symposium/Convention in Vancouver B.C. Our workshop is titled: > > Resin Applications Forum: Methods for Processing, Special Staining, > Immunohistochemical and In Situ Hybridization of Soft and Hard Tissue > Including Medical Device Implants > > During the last 50 years, numerous histological procedures have been > described on resin embedded tissue. While different types of resins are > available for different purposes, the acrylics provide the widest range of > techniques, especially for light microscopy applications. However, as > demand from H&E to more sophisticated techniques increases, so too have the > problems, and nowhere is this more apparent and controversial than in the > application of immunohistochemistry on resin sections. This workshop will > provide a review and discussion for those individuals that currently work > with and/or are just getting started working with soft and hard tissue > specimens and specifically the various resins (i.e. MMA, GMA, Technovit, > Acrylosin, etc.) associated with their specific tissue interests. The > workshop will also detail the preparation and staining of sections of soft > and hard tissue, including implants (e.g. undemineralized bone and > cardiovascular stents), for immunohistochemical and in situ hybridization > staining using different acrylic and epoxy resin embedding media. Specific > problems and pitfalls, either technical or operational associated with > certain resin embedding procedures, will be illustrated and examined. > Particular emphasis will be given to procedures which have been used > extensively for routine diagnostic, and research purposes, i.e. those that > WORK! Individuals with a current or future intent to process and cut > undemineralized tissue or tissue containing foreign implant materials using > acrylic or epoxy resins are strongly encouraged to attend this workshop! > > Please feel free to contact me if you would like more information about > the workshop as information relevant to the exact date and time becomes > available. All I know at this time is that the NSH meeting is September > 29th - October 3rd, 2012. > > Best Regards, > > Jack > > > Jack Ratliff > Hard Tissue Histologist > Chairman, Hard Tissue Committee - National Society for Histotechnology > > > > > > From: gayle.cal...@bresnan.net > > To: histonet@lists.utsouthwestern.edu > > Date: Mon, 12 Mar 2012 11:04:20 -0600 > > Subject: [Histonet] Re: undecalcified bone IHC > > > > Jeff, > > > > > > > > It is most certainly possible to do IHC on undecalcifed bone sections > > embedded in PMMA although not the easiest task. Sectioning is done on a > > microtome that is powerful enough to cut the plastic and using tungsten > > carbide knives. The key is total removal of the plastic from MMA embedded > > bone sections to allow antibody/ immunoglobulins to access antigenic > sites. > > Neil Hand has done IHC successfully on PMMA embedded tissues including > > undecalcified bone on 2 to 3 µm thick sections. I think one could cut > > thicker sections at 4 to 5 µm and still be successful. I do not recall > what > > Troiano et al used. > > > > > > > > The following publications will help you and should include protocols, > > although conventional protocols will work according to Hand. > > > > > > > > Blythe D. Hand N et al 1997 J Clin Path 50:45-49. The use of methyl > > methacrylate resin for embedding bone marrow trephine biopsies. > > > > Hand NM et al 1996 Antigen unmasking using microwave heating on formalin > > fixed tissue embedded in methyl methacrylate J Cellular Path 1:31-37 > > > > Jackson P et al. 1996 Amplification of immunocytochemical reactions by > > the catalytic deposition of biotin on tissue sections. J Path > > 170(suppl):23A. This was about tyramide amplification when one gets a > weak > > signal from "conventional" methods. > > > > Hand NM, Church RJ 1998 Superheating using pressure cooking: its use and > > application in unmasking antigens embedded in methyl methacrylate. J > > Histotechnology 2`:231-236 > > > > Hand NM et al 1989 Immunohistochemistry on resin embedded tissue for > light > > microscopy: a novel post embedding procedure. Proceeding Royal > > Microscopical Society 24(1):A54-55. > > > > Hand NM Plastic Embedding media and techniques, Ch.30, p 663-677. Theory > > and Practice of Histological Technique, 5th edition by Gamble and > Bancroft. > > The 6th edition is updated under same title. > > > > > > > > Use Google Scholar to find Troiano N et al from Yale on doing IHC on PMMA > > embedded bone sections with publications in J Histotechnology. > > > > > > > > > > > > Hand mentioned several HIER methods, using citrate buffer. Optimizing > > retrieval will depend on the antigen and you may end up doing this with > some > > form of HIER, including microwave or other heat producing methods and > with > > different buffers. Enzyme digestion is also a possibility. > > > > > > > > Hand removed MMA with xylene, warm my speed up the removal, also more > than > > one change for 10 - 20 minutes or longer. When I talked to him > personally, > > he said he had used warm xylene although temperature was not mentioned in > > his chapter. After MMA removal, rehydrate section through alcohol > gradient > > as one does paraffin sections. He was emphatic about never allowing the > > sections dry out. > > > > > > > > Hopefully Jack Ratliff and Damien Laudier will provide more insight on > this > > topic. > > > > > > > > Good luck > > > > > > > > Gayle M. Callis > > > > HTL/HT/MT(ASCP) > > > > > > > > > > > > > > > > > > > > > > > > > > > > ****************************************** > > > > > > > > Hi Jeff, > > > > > > > > If is it possible a few more specifics of how the tissue has been > received, > > > > processed and evaluated would help. Undecalcified bone sectioning > > > > procedures vary and also what specific markers are you looking to do is > > > > important. > > > > > > > > Vikki > > > > On Mon, Mar 12, 2012 at 11:06 AM, Rene J Buesa <rjbuesa <@t> yahoo.com> > > wrote: > > > > > > > > > Undecalcified? How are you going to section it? > > > > > If you can section it, just use any IHC protocol for regular sections. > > > > > Good luck! > > > > > René J. > > > > > > > > > > --- On Mon, 3/12/12, Jeffery Howery <Jeffery.Howery <@t> jcl.com> > wrote: > > > > > > > > > > > > > > > From: Jeffery Howery <Jeffery.Howery <@t> jcl.com> > > > > > Subject: [Histonet] Undecalcified bone IHC > > > > > To: histonet <@t> lists.utsouthwestern.edu > > > > > Date: Monday, March 12, 2012, 10:59 AM > > > > > > > > > > > > > > > Does anyone have a protocol for Undecalcified bone for IHC? > > > > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Damien Laudier Laudier Histology www.LaudierHistology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet