[Histonet] donation request
Dear Colleagues, We are expecting a quite busy period in terms of research studies. As some of you do also experiencing similiar issues, we are using fluorescent microscopes from other depts when available. But mostly it is not easy. Buying a new one does not seem possible soon due to financial issues. My question is, if any colleague has a redundant fluorescein microscope available to donate us. Your kindly helps would be greatly appreciated. Thanks in advance. Dr. Necat Yilmaz MD, PhD University of Mersin School of Medicine Histology and Embryology Dept. Mersin/TURKEY ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Snap freezing_method by John Tarpley_reg
Dear Histonetters, This query is with regard to snap freezing technique of rodent liver for cryotomy. Thanks a lot for the extensive details available with our forum especially from Gayle Callis. But I have seen numerous quotes regarding the use of the method from John Tarpley, who published his method in the magazine microscopy today, june 2001 issue. This issue is no longer available and I have already written to Cambridge publishers. In the mean time, if any of our histotechs having this article with them to share me a copy or any body able to give me the contact details of the author John Tarpley, so that I can contact him for a reprint Thanks for all your help Abijag Love Cricket? Check out live scores, photos, video highlights and more. Click here http://cricket.yahoo.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Preventing livers from falling apart after dab stain
HI: I have a = question regarding keeping livers intact. I am currently removing o= rgans of dosed animals and freezing them in OCT. I cryostat these livers = in 40um sections and are kept in -80C until used. We fix thes= e using 4% Paraformaldehyde, and we use antibodies and DAB for histochemist= ry. What I have been seeing is that my tissue almost looks expanded. Sm= all tissue gaps are magnified and it looks as if chunks of the tissue just = dissapear leaving gaps in between what looks like cell clusters. Th= is is not there when I cut or during -80 storage because I check the slides before and after freezin, and fixation. I would greatly appreciate= some input on how to prevent this from happening. Tha= nks Sean Taheri = This e-mail and any attachment hereto, is intended= only for use by the addressee(s) named above and may contain legally privi leged and/or confidential information. If you are not the intended recipi= ent of this e-mail, any dissemination, distribution or copying of this emai= l, or any attachment hereto, is strictly prohibited. If you receive this = email in error please immediately notify me by return electronic mail and p= ermanently delete this email and any attachment hereto, any copy of this e-= mail and of any such attachment, and any printout thereof. Finally, pleas= e note that only authorized representatives of Regeneron Pharmaceuticals, I= nc. have the power and authority to enter into business dealings with any third party. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] MMP-2 antibodies
I am looking for some advice as to which MMP-2 antibody people recommend for staining human tissue-either FFPE or frozen sections. I have tried one from Novus as well as Serotec with no luck. I looked in the Archives but couldn't find any. Any suggestions greatly appreciated. Mark abi jag abija...@yahoo.co.in 7/21/2009 5:10 AM Dear Histonetters, This query is with regard to snap freezing technique of rodent liver for cryotomy. Thanks a lot for the extensive details available with our forum especially from Gayle Callis. But I have seen numerous quotes regarding the use of the method from John Tarpley, who published his method in the magazine microscopy today, june 2001 issue. This issue is no longer available and I have already written to Cambridge publishers. In the mean time, if any of our histotechs having this article with them to share me a copy or any body able to give me the contact details of the author John Tarpley, so that I can contact him for a reprint Thanks for all your help Abijag Love Cricket? Check out live scores, photos, video highlights and more. Click here http://cricket.yahoo.com ***CONFIDENTIALITY NOTICE*** This electronic message is intended only for the use of the addressee and may contain information that is privileged and confidential. Any dissemination, distribution or copying of this communication by unauthorized individuals is strictly prohibited. If you have received this communication in error, please notify the sender immediately by reply e-mail and delete the original and all copies from your system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Celloidin Sections.
Hi Barry, I am studying the development of atherosclerotic lesions in mouse brachiocephalic artery--I typically use 5 micron sections. Paraffin sections have given me mostly good, frequently excellent and occasionally spectacular results, morphology-wise. Oh, and occasionally poor results, too. I would prefer to increase the ration of spectacular/merely OK results. I read an article where the authors describe getting better fine morphology with celloidin than paraffin. Atheromas, like cochlea are delicate structures with fine detail, and they can easily become detached from the artery wall. After reading the article, I can't help but wonder if a different embedding media might give me even better results than I am already getting. Would I need a special microtome for celloidin? One reason I don't want to go with methacrylate is I don't have budget for new equipment right now. Jerry Ricks Research Scientist University of Washington Department of Pathology Effects of Fixative and Embedding Medium on Morphology and Immunostaining of the Cochlea Audiol Neurootol. 2009 ; 14(2): 78–87 From: barry.r.ritt...@uth.tmc.edu To: histonet@lists.utsouthwestern.edu Date: Mon, 20 Jul 2009 19:14:58 -0500 Subject: RE: [Histonet] Celloidin Sections. Jerry depends on whether you are talking about celloidin or Low Viscosity Nitrocelulose. There are techniques in the literature to cut celloidin sections as thin as 4 microns. For LVN you need much thicker sections as this plastic is much less sturdy and has a tendency to fragment if too thin. The questions I ask is why you need celloidin sections and what tissue are you thinking about for this technique? Barry From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of JR R [rosenfeld...@hotmail.com] Sent: Monday, July 20, 2009 6:42 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Celloidin Sections. I am curious--How thin can celloidin sections be cut? I've heard that 30 microns is a standard thickness. No way to make 5 micron sections? Thanks, Jerry Ricks Research Scientist University of Washington Department of Pathology _ Windows Live™ SkyDrive™: Store, access, and share your photos. See how. http://windowslive.com/Online/SkyDrive?ocid=TXT_TAGLM_WL_CS_SD_photos_072009___ 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 _ Bing™ brings you maps, menus, and reviews organized in one place. Try it now. http://www.bing.com/search?q=restaurantsform=MLOGENpubl=WLHMTAGcrea=TXT_MLOGEN_Local_Local_Restaurants_1x1___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] A web site for tissue arrays and other sections on slides
I recently came across the web site of a company that provides slides with sections of a wide variety of animal tissues - normal, pathological, transgenic etc: single and multiple tissues, as mounted paraffin sections. The associated photos indicate high quality fixation and sectioning. There are frequent Histonet enquiries about sources of negative and positive controls for immunohistochemistry, special stains etc. This might be such a source. The address is http://www.histobest.com John Kiernan Department of Anatomy Cell Biology University of Western Ontario London, Canada. = = = ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: double immunostaining with antigen retrieval and no antigen retrieval
Cathy, You absolutely can do this, but you have to use a permanent, stable end product on the first antibody staining. This is easy to do if you use DAB as the first staining technique chromogen. The antigen retrieval will probably remove all antibody-antigen bonds (elution) from your first IHC reaction, so having a permanent label on the first one is critical. Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] removing coverslip from immuno slides
Yes, you can use the same procedure. René J. --- On Tue, 7/21/09, Jennifer Campbell jcampb...@vdxpathology.com wrote: From: Jennifer Campbell jcampb...@vdxpathology.com Subject: [Histonet] removing coverslip from immuno slides To: histonet@lists.utsouthwestern.edu Date: Tuesday, July 21, 2009, 3:30 PM I coverslipped some immuno slides using our Tissue-Tek coverslipper and I need to lift the coverslips from a few of the slides (used the wrong length). For histo slides, we typically soak the slides in acetone until the coverslipping film is easily removed. Is it ok to soak immuno slides in acetone to remove the film? Thank you, Jennifer ___ 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] Fluorescent dyes and xylene
Most of the Alexa Fluor dyes appear to be trade secrets: only a few are identified with chemical structures in the Molecular Probes/Invitrogen catalogue. Fluorescent labels of known identity do survive dehydration, clearing and mounting in a permanent medium and are still OK years later, stored in boxes at room temperature. It is, of course, necessary to use a non-fluorescent mountant such as DPX or one of the poly(methacrylate) ones (Entellan, Cytoseal etc). The notion that it's necessary to mount fluorescent preparations in slightly alkaline aqueous media may derive from an influential book by Nairn (1976), which stressed the effects of pH on fluorescence intensity and recommended mounting only in a somewhat alkaline glycerol-water mixture, and storage of slides in flat trays in a fridge. In fact, fluorescent labels bound to proteins survive dehydration, clearing and permanent mounting (see, eg, Stoddart 1973, Whyte 1978, Allen 1994, Espada 2005, Hertzler 2006). The paper by Espada et al (2005) illustrates this point very well and also discusses reasons why a polystyrene medium (ie DPX) may be best. You would not expect dehydration etc to remove or damage a fluorescently labelled antibody or other protein. Immersing the stained section in alcohol coagulates the protein instantly, along with the covalently attached fluorochrome molecules. References. Allen DT, Kiernan JA (1994) Permeation of proteins from the blood into peripheral nerves and ganglia. Neuroscience 99: 755-764. http://www.ncbi.nlm.nih.gov/pubmed/8008217?ordinalpos=33itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_DefaultReportPanel.Pubmed_RVDocSum Espada J, Juarranz A, Galaz S, Canete M, Villanueva A, Pacheco M, Stockert JC (2005) Non-aqueous permanent mounting for immunofluorescence microscopy. Histochem. Cell Biol. 123: 329-334. http://journals1.scholarsportal.info/details.xqy?uri=/09486143/v123i0003/329_npmfim.xml Hertzler PL (2006) Rhodamine fluorescence after 15-year storage in methyl salicylate. Microscopy Today 14-2: 48. http://www.microscopy-today.com/jsp/mto/print_archive/print_archive.faces Nairn RC (1976) Fluorescent Protein Tracing. 4th ed. Churchill-Livingstone, London. ISBN 0443012733. http://openlibrary.org/b/OL5063508M/Fluorescent-protein-tracing Stoddart RW, Kiernan JA (1973) Histochemical detection of the alpha-D-arabinopyranoside configuration using fluorescent-labelled concanavalin A. Histochemie 33: 87-94. http://www.springerlink.com/content/vk667214g6m80530/?p=fb8574a99977492fa339fb2300500e29pi=2 Whyte A, Loke YW, Stoddart RW (1978) Saccharide distribution in human trophoblast demonstrated using fluorescein-labelled lectins. Histochem. J. 10: 417-423. http://www.springerlink.com/content/h710461877mk8010/ John Kiernan Department of Anatomy Cell Biology University of Western Ontario London, Canada. = = = - Original Message - From: Bob Nienhuis bob.nienh...@gmail.com Date: Monday, July 20, 2009 14:10 Subject: [Histonet] Fluorescent dyes and xylene To: Histonet histonet@lists.utsouthwestern.edu Will the Alexa Fluor dyes withstand some dehydration and clearing in alcohol and xylene? If they won't are there any bright fluorescent dyes that can be obtainedcongugated to a secondary antibody that will? Looking to immunostain tyrosine hydroxylase in rodent brain with a fluorescent marker in tissue that needs to be dehydrated and cleared, perhaps with an abbreviated series. Bob Nienhuis VA / UCLA Neurobiology Research ___ 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] Average pay rate southern cali
I'm looking for an employee in southern california to be a lead tech for a derm lab. Just wondering what the histonet sees as far as pay rate for a tech with some experience. Please give me some hypothetical ballpark figures for a certified tech as well as uncertified, keeping in mind this person will do all routine work plus some administrative work. Thanks! Sent from my Verizon Wireless BlackBerry ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
