From Lawrence.Brett <@t> luht.scot.nhs.uk Fri Oct 1 03:56:34 2004 From: Lawrence.Brett <@t> luht.scot.nhs.uk (Brett, Lawrence) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Factor 13a Message-ID: <857344E7D8F90240935A288998A89D811C605A@wgh-ex1.luht.scot.nhs.uk> Martha We use factor 13a from Neomarkers - clone AC-1A1, order code MS-1237-PO In the UK they are disributed by LabVision - don't know about US. It works well. Lawrence Brett Edinburgh Scotland, UK ********************************************************************** The information contained in this message may be confidential or legally privileged and is intended for the addressee only, If you have received this message in error or there are any problems please notify the originator immediately. The unauthorised use, disclosure, copying or alteration of this message is strictly forbidden. ********************************************************************** From Marion.Hiles <@t> north-bristol.swest.nhs.uk Fri Oct 1 08:22:07 2004 From: Marion.Hiles <@t> north-bristol.swest.nhs.uk (Marion Hiles) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Barbital Message-ID: <2EE924DF60902943AC6E2EF35155451F19CD37@nbfexch03.north-bristol.nhs> Very interested in your substitute for Barbitol. In the incubating medium, you wrote 10mg of 0.1M glycine buffer with calcium chloride is added, surely its a liquid?! Bob Quilty Neuropathlogy Frenchay Hospital Bristol UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bonner, Janet Sent: 29 September 2004 18:35 To: 'Vicki Gauch'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Barbital We use 0.1M Glycine Buffer with 0.75M Calcium Chloride: 0.1M Glycine Buffer: Glycine...........750 mg Sodium chloride...585 mg Distilled water...100 ml Stable for 6 months at 4c 0.75M Calcium Chloride, Dihydrite: Calcium Chloride, dihydrite....11.03 gm Distilled water...............100 ml 0.1M Sodium Hydroxide: Sodium Hydroxide.........0.4gm ____________ Distilled water..........100 ml 0.1M Glycine Buffer with 0.75M Calcium Chloride 0.1M Glycine buffer..........50 ml 0.75M Calcium Chloride........10 ml 0.1M Sodium hydroxide........22 ml Adjust pH to 9.4 with additional sodium hydroxide as necessary. As the pH increases, the calcium precipitates. Let the precipitate settle to the bottom or filter before use. May be stored for 6 months at 4C. Warm to room temperature and check pH before use. Incubation Solution: Adenosine triphosphate(ATP)................5 mg 0.1M glycine buffer with calcium chloride..10 mg Mix and adjust pH to 9.4 with 0.1M sodium hydroxide or 0.1M hydrochloric acid. Our main reference is Frieda Carson's HISTOTECHNOLOGY:A SELF INSTRUCTIONAL TEXT, second edition, pp 260-262, pp264-265. -----Original Message----- From: Vicki Gauch [mailto:GauchV@mail.amc.edu] Sent: Wednesday, September 29, 2004 11:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Barbitol Does anyone know what we can substitute for Barbitol in ATPase buffers for muscle stains? Any help you can give me would be greatly appreciated.. Thanks, Vicki Gauch AMCH Albany, NY _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet DISCLAIMER: The information in this message is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorised. If you are not the intended recipient, any disclosure, copying, or distribution of the message, or any action or omission taken by you in reliance on it, is prohibited and may be unlawful. Please immediately contact the sender if you have received this message in error. Thank you. From ploykasek <@t> phenopath.com Fri Oct 1 09:38:18 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Permanent Red Message-ID: Just a quick question on alternative chromogens. Has anyone had a problem with Dako?s Permanent Red coming out in recycled alcohols? This is a nice alk. Phos. Chromogen, and I really like it. Occasionally though, it comes out in the alcohols in our counterstaining/dehydrating set up ? not every time. We do use recycled alcohols in this set-up. If we switch the recycled out for all fresh, we don?t have this problem. Just wondering. Patti Loykasek PhenoPath Laboratories Seattle, WA From Luis.Chiriboga <@t> med.nyu.edu Fri Oct 1 10:05:31 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] NSH in ADVANCE for Medical Laboratory Professionals Editorial Message-ID: Click on the link for synopsis of NSH meeting http://www.laboratorian.advanceweb.com/common/Editorial/Editorial.aspx?CC=41 391 From JKOBLER <@t> PARTNERS.ORG Fri Oct 1 10:06:53 2004 From: JKOBLER <@t> PARTNERS.ORG (Kobler, James) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] big specimen Message-ID: <57531340B9FDD611A8580008026158F1096EF5AE@phsexch26.mgh.harvard.edu> I need help cutting a large specimen - the partially dissected larynx of a dog. This specimen, currently in 10% formalin, is roughly the size of an egg. We need some nice whole mount H&E-stained sections through it in a coronal plane. If someone could process this for us in an expeditious manner, please contact me: Jim Kobler, jkobler@partners.org , 1-617-726-0212. Thanks very much. Jim Kobler, Department of Surgery, Massachusetts General Hospital, Boston. From cwscouten <@t> myneurolab.com Fri Oct 1 10:08:52 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Sucrose cryoprection Message-ID: A few drops of a strong base are usually added to clear the paraformaldehyde into solution after it is warmed up. Look up a recipe for this. You may not be getting a perfusion, the fluid my be flowing into and out of the heart, with the capilaries blocked by powder or bubbles or red blood cells. Try a high pressure perfusion. See the following link: http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=471001&catdesc=Histology+Equipment&CatThreeID=674&CatOneID=4&subcatdesc=Sacrifice+Equipment&idsubcategory=21 Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pablo S?nchez Quinteiro Sent: Thursday, September 30, 2004 2:18 PM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sucrose cryoprection Thanks to everybody who has made me on- and off-list useful and interesting hints. Now it seems clear to me that I have a problem with fixation. I'll try to be more careful; for example making prewash (I do not do that in order to simplify the complicated procedure of perfusing so tiny animals) or checking if there is powder of Paraformaldehyde in the botton (Some times a few grains remain in the bottom). I think that in addition to this the postfixation time must have the key role. By the way I would like to know your experience with nervous tissue in young animals (1-7 days old in mouse, for example). It is not easy to fix properly this postnatal animals. When I perfuse the paraformaldehyde in adult animals I can see the sad limbs movements which indicate that fixation is going well. In postnatal animals I never get this effect. Have you got any idea regarding this difference? Maybe the more water content in postnatal animals or perhaps -as cleverly a co-lister has suggested me- the more immature CNS system in postanatal animals. Thanks again Pablo X-Virus-Scanned: by usc.es X-MIME-Autoconverted: from quoted-printable to 8bit by secuslugo.lugo.usc.es id i8UEd9O26589 It does sound like the tissue is not fixed, although 48 hours in Paraformaldehyde should be sufficient even without perfusion. Is the solution fully formed? Did any white powder remain in the bottom? It seems you are not getting good fixative. Did you prewash with saline or sucrose, or just perfuse? Blood can block many capillaries and block a through perfusion. Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ajennings <@t> unmc.edu Fri Oct 1 10:11:34 2004 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Histonet @ NSH S/C In-Reply-To: Message-ID: I don't want to step on any toes, I have just recently returned to the listserv, but I do not want this opportunity to pass. Someone please step forward and take command, I will do everything I can to help. Kerry Please let us know when someone accepts the task. kerry.l.crabb@gsk .com Sent by: To histonet-bounces@ histonet@lists.utsouthwestern.edu lists.utsouthwest cc ern.edu Subject [Histonet] Histonet @ NSH S/C 09/30/2004 07:12 PM All, I appreciate your interest in the NSH Symposium/Convention and Histonet which has done so much for the histology field. I have been copied on most if not all the emails to Histonet about trying to get you all together during the S/C. Yes, that would be a great idea I believe NSH could support. Yes, it was done a number of years ago (I think twice) during Histonet's early days. Since Histonet is not an official part of NSH, a gathering for you netters is not planned or arrangements made by NSH. Although NSH does not have monies available to assist you, we may be able to assist by finding space for you to gather if someone in your group is willing to plan and make all the arrangements. The person willing to plan something for you may contact the NSH Meeting Manager (Carrie Diamond) to see if she can find a table/room for you to gather. This person may also contact me directly to discuss your ideas as they develop and the possibilities of making them happen at the next NSH S/C. Kerry Crabb NSH Convention Chairman _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dobbin <@t> upei.ca Fri Oct 1 11:06:21 2004 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Histonet @ NSH S/C In-Reply-To: References: Message-ID: <415D564D.17032.F244CD@localhost> Hello Everyone, I do plan to move this Histonet Outpost idea forward for the next NSH S/C. I appreciate all the offers to help out. I will undoubtly be contacting all of you, especially since I am not too confident I'll be able get funds to attend the meeting myself! Kerry, How soon should I start making arrangements? I had been thinking about waiting until the new year. Maybe that is not soon enough? Greg Copies to: histonet@lists.utsouthwestern.edu From: ajennings@unmc.edu Date sent: Fri, 1 Oct 2004 10:11:34 -0500 Subject: Re: [Histonet] Histonet @ NSH S/C > > > > > I don't want to step on any toes, I have just recently returned to the > listserv, but I do not want this opportunity to pass. > Someone please step forward and take command, I will do everything I can to > help. Kerry Please let us know when someone accepts the task. > > > > > kerry.l.crabb@gsk > .com > Sent by: To > histonet-bounces@ histonet@lists.utsouthwestern.edu > lists.utsouthwest cc > ern.edu > Subject > [Histonet] Histonet @ NSH S/C > 09/30/2004 07:12 > PM > > > > > > > > > All, > > I appreciate your interest in the NSH Symposium/Convention and Histonet > which has done so much for the histology field. I have been copied on > most if not all the emails to Histonet about trying to get you all > together during the S/C. Yes, that would be a great idea I believe NSH > could support. Yes, it was done a number of years ago (I think twice) > during Histonet's early days. Since Histonet is not an official part of > NSH, a gathering for you netters is not planned or arrangements made by > NSH. Although NSH does not have monies available to assist you, we may be > able to assist by finding space for you to gather if someone in your group > is willing to plan and make all the arrangements. The person willing to > plan something for you may contact the NSH Meeting Manager (Carrie > Diamond) to see if she can find a table/room for you to gather. This > person may also contact me directly to discuss your ideas as they develop > and the possibilities of making them happen at the next NSH S/C. > > Kerry Crabb > NSH Convention Chairman > _______________________________________________ > 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 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Pathology Lab Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Happiness is a journey, not a destination. From sluhisto <@t> yahoo.com Fri Oct 1 11:13:48 2004 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] enos Message-ID: <20041001161348.80649.qmail@web51004.mail.yahoo.com> Hello All: I have an investigator that is looking to do IHC using VegeF and Enos antibodies on human lung tissue. Does anyone out there have vendors, protocols, dilutions, or any information that you might share with her. Thanks, in advance for your help. Susan --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From ljb <@t> medicine.wisc.edu Fri Oct 1 11:12:17 2004 From: ljb <@t> medicine.wisc.edu (LaCinda Burchell) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] florida hurricanes Message-ID: May God Bless You all in Polk County, (if people want to flame me for that expression, so be it) One hurricane was a disaster, I can't imagine living through three! Is the Red Cross still our best bet for getting help down to you? Take Care, and hold tight to your loved ones. Cindy B LaCinda Burchell, BA, AS, HT(ASCP) University of Wisconsin-Madison, Medical School Asthma and Allergy Research IHC Lab 600 Highland Ave. CSC K4/913 Madison, Wisconsin 53792 Phone: 608-262-3518 FAX: 608-263-3746 >>> 09/30/04 08:04PM >>> Hello everyone. I know this isn't the venue to post things like this, but it is much easier for me to write one email instead of sending several. First, I want to say that I am sorry that I was unable to go to Nationals. I know that several of you were counting on me. Sharon told me that you were concerned and asking about me at the Convention. Let me say that I am ok. Not great, but OK. I did get hit by 3 of the 4 hurricanes that went through Florida these past 6 weeks. Charley, Francis, and Jeanne all went directly over the county that I live in. Once things started to get taken care of from one hurricane, another one would come. The last one, Jeanne, came around midnight on Saturday night and my power didn't come back on until today. Hopefully it will stay on. There still are thousands of people without homes or power. As of today, the kids have missed 16 days of school already. It has been extremely stressful, but I am one of the lucky ones because I still have a home. The news reports that you may have seen do not even come close to the destruction here, not even close. A lot of businesses were destryoed as well. But organizations are helping with food, water, ice, shelter, etc. And prayers from everyone help too. Just keep praying for all of us in Polk County Florida......And that you for your concerns. I will see you next year. Roxanne Soto _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pzeitlow <@t> bbpllab.com Fri Oct 1 11:20:41 2004 From: pzeitlow <@t> bbpllab.com (Pat Zeitlow) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] CAS200 Image Analyzer Message-ID: <813FB33DA405334F947F8BFC6EBD0B2A0BEA9D@bbplsrv1.bbpl> We are in the process of dismantling our CAS 200 which has been replaced. I know parts are very hard to come by. If anyone is interested in parts, etc please contact me directly via email or phone (573-886-4689). Pat Z Department of Molecular Pathology Boyce and Bynum Pathology Labs, P.C. From neuroant <@t> hotmail.com Fri Oct 1 11:29:21 2004 From: neuroant <@t> hotmail.com (Ant S.) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Solochrome Cyanide R Message-ID: Does anyone out there know about Solochrome Cyanide R stain for myelin staining? Please tell me where we could find out about the stain, and if you have any tips or recipes you could share, that would be super nifty. Your help is appreciated, thanks. Antoinette Swensson Univeristy of Washington/Harborview Medical Center Neuropathology _________________________________________________________________ [1]Express yourself instantly with MSN Messenger! Download today - it's FREE! References 1. http://g.msn.com/8HMBENUS/2728??PS=47575 From mward <@t> wfubmc.edu Fri Oct 1 12:31:08 2004 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] FXIIIa antibody Message-ID: <61135F0455D33347B5AAE209B903A304076A4F73@EXCHVS2.medctr.ad.wfubmc.edu> Thanks to everyone who responded to my question about the FXIIIa. Martha Ward From eenriquez52234 <@t> yahoo.com Fri Oct 1 12:37:57 2004 From: eenriquez52234 <@t> yahoo.com (esteban enriquez) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Tissue Processing Message-ID: <20041001173757.92380.qmail@web61110.mail.yahoo.com> Does any know if the new released Visiun Biosystem tissue processor can use a generic petroleum distillate as cleaner ?? The web page doesn't say nothing to this . This would be a big saving for money. Esteban __________________________________ Do you Yahoo!? Yahoo! Mail is new and improved - Check it out! http://promotions.yahoo.com/new_mail From sluhisto <@t> yahoo.com Fri Oct 1 12:42:16 2004 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] von leder Message-ID: <20041001174216.69622.qmail@web51010.mail.yahoo.com> Does anyone have a protocol or suggestions for doing a von Leder stain for neutrophils in mouse lung? Any and all help is greatly appreciated. Susan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From gcallis <@t> montana.edu Fri Oct 1 13:17:37 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Solochrome Cyanide R In-Reply-To: References: Message-ID: <6.0.0.22.1.20041001121621.01b30bf8@gemini.msu.montana.edu> It is called solochrome cyanine R, it stains the myelin black, and very effective. John Kiernan will probably reply as he supplied me with a publication AND method, which worked very nicely. I will attach my staining method to you separately. At 10:29 AM 10/1/2004, you wrote: > Does anyone out there know about Solochrome Cyanide R stain for myelin > staining? Please tell me where we could find out about the stain, and > if you have any tips or recipes you could share, that would be > super nifty. > > Your help is appreciated, thanks. > Antoinette Swensson > Univeristy of Washington/Harborview Medical Center > Neuropathology > _________________________________________________________________ > > [1]Express yourself instantly with MSN Messenger! Download today - > it's FREE! > >References > > 1. http://g.msn.com/8HMBENUS/2728??PS=47575 >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From crhoyle <@t> hotmail.com Fri Oct 1 13:50:38 2004 From: crhoyle <@t> hotmail.com (Cornelia Smith) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] YB-1 Message-ID: Dear Histonetters, Can anyone point me to a vendor for YB-1 antibody that performs on FFPE? Thanks in advance! CR Smith _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From jkiernan <@t> uwo.ca Fri Oct 1 13:57:06 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Solochrome Cyanine R References: Message-ID: <415DA882.8EF18C1D@uwo.ca> First, the name! In most catalogs this dye is called eriochrome cyanine R, so that's the name used in the current (10th) edition of Conn's Biological Stains. Two synonyms, much used in the literature, are solochrome cyanine R (ICI's trade-name, I think), and chromoxane cyanine R (which originated, I think, with some pre-war German company and was used in the 9th edn of Conn's). The Colour Index name is CI Mordant blue 3; CI 43820. The dye has several uses in histology and there are several published procedures for staining myelin. My favourite is as follows: __________________________________________________________________ Iron-eriochrome cyanine R for myelin (after Kiernan, 1984; original was Page, 1965). Staining solution. 0.21 M aqueous ferric chloride (5.6% w/v FeCl3.6H2O): 20.0 ml Eriochrome cyanine R (C.I. 43820): 1.0 g Concentrated (95-98% w/w) sulphuric acid: 2.5 ml Water: to make 500 ml (The ingredients take 2-5 minutes to dissolve, with stirring. Filtration should not be necessary. The solution can be kept and used repeatedly for at least 8 years.) Differentiating solution for myelin. Any one of the following aqueous solutions may be used. See also Note below. Iron alum (NH4Fe(SO4)2.12H2O): 10% w/v Ferric chloride (FeCl3.6H2O): 5.6% w/v Ferric nitrate (Fe(N03)3.6H2O): 7.3% w/v (The differentiating solution keeps for a few years, but may be used only once. Discard if it is cloudy or if there is a thick layer of pale insoluble material in the bottom of the bottle.) Counterstain. After a myelin stain, use a red basic dye (0.5% aqueous neutral red or safranine is suitable) to stain nuclei and Nissl substance. Method (paraffin sections of formaldehyde-fixed animal tissues, especially CNS). 1. Stain hydrated sections for 15-20 minutes are needed. (The slides may remain in the dye solution for 30 minutes without harm.) 2. Wash in running tap water, 30 seconds, or in 3 changes of distilled water. This is to remove unbound dye. 3. Immerse in the differentiating solution until only the myelin (white matter of CNS) retains the stain. This usually takes 5-10 minutes. It is sometimes impossible to decolorize the nuclei completely without losing some intensity in myelin. 4. Wash in tap water (running, or three or four changes) for about 5 minutes. 5. Apply a counterstain, as desired. 6. Dehydrate, clear, and cover using a resinous mounting medium. Result. Myelin and red blood cells dark blue. ________ Note on differentiation. Clark (1979) recommended an alkaline differentiating solution for myelin staining: a freshly prepared 1% (v/v) dilution of ammonium hydroxide. This acts in a few seconds, and it is easy to remove too much of the blue dye-metal complex. ________ References. Clark, G. (1979). Staining with chromoxane cyanine R. Stain Technology 54: 337-344. Kiernan, J.A. (1984). Chromoxane cyanine R. II. Staining of animal tissues by the dye and its iron complexes. Journal of Microscopy 134: 25-39. Page, K.M. (1965). A stain for myelin using solochrome cyanin. Journal of Medical Laboratory Technology 22: 224-225. _________ The colour is darker than myelin that has been stained with a luxol fast blue. If the differentiation (Step 3) is done instead with acid-alcohol, the result is a clean blue nuclear stain similar to that seen with a haemalum but more resistant to extraction by subsequently applied acidic counterstains. ____________________________________________________________ -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ "Ant S." wrote: > > Does anyone out there know about Solochrome Cyanide R stain for myelin > staining? Please tell me where we could find out about the stain, and > if you have any tips or recipes you could share, that would be > super nifty. > > Your help is appreciated, thanks. > Antoinette Swensson > Univeristy of Washington/Harborview Medical Center > Neuropathology > _________________________________________________________________ > From Dorothy.L.Webb <@t> HealthPartners.Com Fri Oct 1 14:11:45 2004 From: Dorothy.L.Webb <@t> HealthPartners.Com (Dorothy.L.Webb@HealthPartners.Com) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] alcoholic formalin Message-ID: Would someone please give me the recipe for alcoholic formalin to be used on the second station of a tissue processor, I believe a 70% mixture? I would like to do some experiments with it on one of my processors and I am not certain if there are any additives besides formaldehyde and alcohol and to what proportions each. Thanks much! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From jkiernan <@t> uwo.ca Fri Oct 1 14:20:32 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:06 2005 Subject: Fuchsine (Was: [Histonet] Red chromogens [alkaline phosphatase labels]) References: <000001c4a68d$03c24780$c211a6a5@renad4yk9b8abe> <415B7D83.64C14C0A@uwo.ca> <001c01c4a6d1$7acdf470$eeeea8c0@server> Message-ID: <415DAE00.E56C7AAE@uwo.ca> Point taken! I notice that Chroma do use the correct English spelling of fuchsine. Like nearly everyone else they spell phloxin with a terminal e, which is wrong because it's in the same group as eosin. Chroma also put a wrong terminal e on their English spelling of erythrosin; that one is unusual. --- John Kiernan ------------------------------------ Gudrun Lang wrote: > > Dear John, > I think the German spelling is Fuchsin (without e). Please look at > www.chroma.de at their catalogue. > Your vixen is German "F?chsin" (fuechsin). > > greetings from Austria > Gudrun > > ----- Original Message ----- > From: "John Kiernan" > To: "Rena" ; > Sent: Thursday, September 30, 2004 5:29 AM > Subject: Re: [Histonet] Red chromogens [alkaline phosphatase labels] > > Dear Rena Fail, > > Red alkaline phosphatase product. > > Methods giving permanently mountable red products have been available > for many years. > Davis & Ornstein (1959) introduced hexazotized pararosaniline as a > trifunctional trapping agent for naphthols released by hydrolysis of > naphthol phosphates. Pararosaniline (CI 42500; CI Basic red 9) is one of > the four components of basic fuchsine and is commercially available in > fairly pure form. New fuchsine (CI 42520; CI Basic violet 2), which is > closely similar to pararosaniline and also available in pure form > (Horobin, 2002); It is currently preferred for making the hexazotized > reagent. > > Kits are sold, but the substrate solution is easily made in the lab. In > a simple procedure published at the IHCWorld.com (2004) web site (see > below for its URL), the working mixture is prepared from four stock > solutions, which are stored at 4 C. Solution D is warmed to room > temperature before using. The amount of levamisole (inhibitor of > endogenous tissue alkaline phosphatase) is clearly stated. > > The working solution is made immediately before using. Mix 10 drops each > of A and B, then add 10 drops of solution C followed by 10 Kits are > sold, but the substrate solution is easily made in the lab. In a simple > procedure published at the IHCWorld.com (2004) web site, the working > mixture is prepared from four stock solutions, which are stored at 4 C. > Solution D is warmed to room temperature before using. I've annotated > the instructions from IHCworld.com to simplify the weighing and > measuring. > > A. New fuchsine (CI 42520), 0.2% in 2M HCl. (Conc. HCl is 10 or 12M) > B. Sodium nitrite (NaNO2), 0.4% and levamisole,(= [-]-tetramisole > hydrochloride) 0.48% in water. > C. Naphthol AS-BI phosphate, 1% in 100% dimethylformamide. > D. 0.05 M Tris-HCl buffer, pH 8.7. > > The working solution is made immediately before using. Mix 10 drops each > of A and B, then add 10 drops of solution C followed by 10ml of the > buffer (D). A drop is assumed to be 0.05 ml. > > Sections are incubated in the working solution for 10-20 minutes at room > temperature. They are then washed counterstained as desired, dehydrated, > cleared and coverslipped. > > Lojda et al (1979, p. 74-75) warn that the working solution may be red > or brown from colored impurities in new fuchsine or pararosaniline. Such > solutions cause excessive yellow background staining. The yellow/brown > impurities are the same ones that can make Schiff's reagent yellow > (removable from Schiff with activated charcoal). Pararosaniline > certified by the Biological Stain Commission is suitable for making > alkaline phosphatase substrate mixtures because it is required to be > substantially free of brown and yellow materials (Penney et al., 2002). > > Probably any batch of basic fuchsine certified by the Biological Stain > Commission will be OK for making the IHCWorld.com alkaline phosphatase > substrate solution, because each of the four possible components of > basic fuchsine has three diazotizable amino groups and their molecular > weights are all quite close. I don't know of any published study that > establishes this, so you're probably safest to go with either certified > pararosaniline or with new fuchsine from a reputable dealer. > > References. (Please check them out if you can. The 2nd is easy enough!) > > Davis & Ornstein (1959) J. Histochem. Cytochem. 7:297-298. > > IHCWorld.com (2004) New fuchsin alkaline phosphatase substrate solution. > http://www.ihcworld.com/_protocols/chromogen_substrates/AP_new_fuchsin_red.htm > > Horobin (2002) Chapter 14 in Conn's Biological Stains, 10th ed. > Oxford:BIOS. > > Lojda et al (1979) Enzyme Histochemistry. Berlin: Springer. > > Penney et al. (2002) Biotech. Histochem. 77:237-275. > > Finally, please note that the correct spelling is fuchsine, not fuchsin, > despite anything you might read in a catalog or on a bottle label. This > is not a matter of American, British and German spellings. (Fuchsin is > German for vixen; the dyes are, I think, named from their colours being > similar to those of Fuchsia flowers rather than foxes.) Look in > Websters, the Oxford, etc. It's fuchsine with an e. An -ine ending of an > informal name of a chemical indicates that it is a base, often an amine. > The -in ending occurs with compounds that are not bases, such as > dextrin, eosin etc. > > John Kiernan > London, Canada > ------------------------------------------------------------------ > Rena wrote: > > > > I have been having some trouble with some Abs stained with New Fuchsin > > and permanent red. Recently I was asked if I were using Levamisole in > > the permanent red and if so how much? I was told too much Levamisole in > > permanent red would result in no staining. I have used 1 drop per ml for > > both New Fuchsin and permanent red, which is the appropriate amount. We > > have run both DABS and either permanent red or New Fuchsin with the > > same AB from the same vial with vastly different results. Any ideas? > > Rena Fail > > _______________________________________________ > > 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 -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ From ajennings <@t> unmc.edu Fri Oct 1 14:22:00 2004 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] antibody needed as control in rat skin In-Reply-To: Message-ID: Tissue: rat skin that has been fixed in formalin and embedded in paraffin. I need a monoclonal antibody (mab) that will work with little background but specific confluent staining without pretreatment. I have tried mab to actin and lamin both from Chemicon and get unacceptable background staining using Vector ABC elite anti mouse and DAB substrate kits for the procedure and visualization. I am testing the same antibodies with a DAKO anti-mouse HRP, but I am interested in knowing if anyone has a 'great' mab that would work with the Vector kits. I do try and clean them up with extended washes and blocks, what I am looking for is a tried and true antibody that stains specifically either nuclear or cytoplasmic doesn't matter, this antibody will be used as a control to test the technical side of immunohistochemical staining procedures. Any suggestions of a mab or that is clean in rat skin? From haldana <@t> unimoron.edu.ar Fri Oct 1 12:15:21 2004 From: haldana <@t> unimoron.edu.ar (Hernan Aldana Marcos) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Re. Kluber barrera stain Message-ID: <000101c4a7f1$ebb6d000$a904a8c0@um.edu> Dear Lynette Elby I send to you our paper dealing to the standardization of the Kluver Barrera stain making in laboratory. Contact me for more details. Sincerelly yours Dr. Hern?n J. Aldana Marcos Facultad de Medicina. Universidad de Mor?n Machado 914. B1708JPD. Buenos Aires. Argentina e-mail alternativo hernanjavier@yahoo.com web: http://hjaldanamarcos.bravepages.com http://histologia.bigthicketdirectory.net/main.html From juan.gutierrez <@t> christushealth.org Fri Oct 1 15:13:11 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] T-Box2 antibody search Message-ID: Y'all best stop making fun of us! Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor San Anttonio, TEXAS (It's like a whole other country) (210)704-2533 -----Original Message----- From: Jackie.O'Connor@abbott.com [mailto:Jackie.O'Connor@abbott.com] Sent: Wednesday, September 29, 2004 4:26 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] T-Box2 antibody search Hi y'all - (sorry, my daughter's boyfriend from Texas was here visiting) Anyway - I'm looking for T-Box2 for IHC - any tips?? Jackie O' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Fri Oct 1 15:19:26 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] slighty off-topic question for IHC workers Message-ID: Other than growing whiskers, no problems. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 -----Original Message----- From: Jan Shivers [mailto:shive003@umn.edu] Sent: Wednesday, September 29, 2004 10:49 AM To: histonet Subject: [Histonet] slighty off-topic question for IHC workers My personal doctor has asked me to pose this question to others in my professional field. I have some interesting medical history I'd like to share with you, and am asking for any comments you may have (and I apologize ahead of time if you think this is an inappropriate forum in which to broach this subject). Four years ago, after a routine health check-up, my bloodwork came back with a positive FANA test. My general practitioner referred me to a rheumatologist for follow-up, saying that I possibly had lupus erythematosis. The subsequent follow-up proved negative, since I had no other signs nor symptoms of the disease, and no other abnormal bloodwork. Then, one month ago, again a routine health check-up came up with abnormal bloodwork results. But this time, the FANA and other CBC values were normal; however, the rheumatoid factor was now TEN times the normal level, indicating I might have rheumatoid arthritis. Again, after a trip to the rheumatologist, the answer was the same... I did not have any other signs or symptoms of RA. So, the rheumatologist is stymied... and his curiosity was piqued when I told him I've been working with antibodies and animal sera for 20 years (and many of those times without gloves, tsk tsk). He's wondering if there might be some passive absorption of antibodies/animal proteins into my system and my body is mounting a wacky response to them. At least this is what I gathered from his query. Has anyone had similar experiences that I can relay to him as anecdotal information? And before you all send those thousands of get-well cards, I'm in very fine health otherwise! Thanks for any input you may have. Jan Shivers IHC Lab U of MN Vet Diag Lab St. Paul, MN, USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Fri Oct 1 15:22:30 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] IHC equipment Message-ID: Ventana's Benchmarks. The best in the business. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 -----Original Message----- From: Baez, Janet [mailto:jbaez@interscopepath.com] Sent: Tuesday, September 28, 2004 6:16 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC equipment It's been 5 years since we did IHC in-house. We use to run our IHC on the old TechMate 1000. We're thinking of bringing IHC back in-house. Any feed back as to the new equipment available. Any info will be greatly appreciated. Thanks. Janet E. Baez Histology Manager Interscope Pathology Medical Group 21114 Vanowen St. Canoga Park, Ca. Tel. 818-992-7848 Fax 818-992-6654 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Fri Oct 1 16:20:33 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] IHC equipment In-Reply-To: Message-ID: Janet, I saw a new machine at the NSH from Biocare Medical called the Nemesis. It holds up to 84 slides, but the neat thing about it is that you can have programs running, and add more slides if you have to just like the Tekmate 1000. I miss my Tekmate because of the availability of adding more slides while the machine is running. Their numbe is 800-799-9499 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of GUTIERREZ, JUAN Sent: Friday, October 01, 2004 3:23 PM To: Baez, Janet; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] IHC equipment Ventana's Benchmarks. The best in the business. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 -----Original Message----- From: Baez, Janet [mailto:jbaez@interscopepath.com] Sent: Tuesday, September 28, 2004 6:16 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC equipment It's been 5 years since we did IHC in-house. We use to run our IHC on the old TechMate 1000. We're thinking of bringing IHC back in-house. Any feed back as to the new equipment available. Any info will be greatly appreciated. Thanks. Janet E. Baez Histology Manager Interscope Pathology Medical Group 21114 Vanowen St. Canoga Park, Ca. Tel. 818-992-7848 Fax 818-992-6654 _______________________________________________ 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 From GREYTRUNK <@t> aol.com Fri Oct 1 16:53:05 2004 From: GREYTRUNK <@t> aol.com (GREYTRUNK@aol.com) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] florida hurricanes Message-ID: <146.3518d611.2e8f2bc1@aol.com> Yes, donations can be sent through the Red Cross, the Salvation Army and several other agencies as well. Thank you Roxanne From lpwenk <@t> sbcglobal.net Fri Oct 1 17:25:12 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] alcoholic formalin References: Message-ID: <004301c4a805$85340be0$5d30d445@domainnotset.invalid> Depending upon the who's procedure you are using, the final alcohol percent is somewhere between 60-80%. The final formalin percent is 10% (= 4% formaldehyde). We make ours up in a discarded plastic one gallon container. One quart is about 0.95 L. So 1 gallon = 4 quarts = 3.8 L = 3800 mL. Figuring it out mathematically, and rounding amounts up or down to make it slightly easier to measure (so our "measurements" are accurate to within about 2.5 mL, but the solution still does it's job): - 2700 mL of 95% alcohol - 905 mL of d. water - 95 mL of 40% formaldehyde (conc) We use the same gallon jug(s) over and over. The easy way to make the alcoholic formalin is to: - measure the 2700 mL of 95% alcohol, pour into gallon container, and mark a line on the container with a black permanent marker, and label this line "95% alcohol" - then measure and add the 905 mL of d. water, and again mark and label the line for "d. water". - finally measure and add the 95 mL of 40% formaldehyde, and again mark and label the line as "40% formaldehyde". After that, when the jug is empty, grab the jug, and fill up each solution to the marked line, eye-balling it. No more graduated cylinders needed. Fast and easy, and though not as accurate, it will be off by only a couple of mL, which may change the percents a fractional amount. Still within good working parameters. One word of warning - make certain it is concentrated 40% formaldehyde. If you start with 10% formalin (4% formaldehyde), and then dilute it another 10%, you will end up with 1% formalin (0.4% formaldehyde), which is next to worthless for fixing in our rush-rush lab environment. Also, use unbuffered. If there are buffering salts, the salts will precipitate out in the alcohol (been there, done that once - whoops). One other thought - why not use it on both stages of the tissue processor? Fixes and starts to dehydrate at the same time. Something else to play around with. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Friday, October 01, 2004 3:11 PM Subject: [Histonet] alcoholic formalin > > Would someone please give me the recipe for alcoholic formalin to be used on > the second station of a tissue processor, I believe a 70% mixture? I would > like to do some experiments with it on one of my processors and I am not > certain if there are any additives besides formaldehyde and alcohol and to > what proportions each. Thanks much! > > ________________________________________ > > This e-mail and any files transmitted with it are confidential and are > intended solely for the use of the individual or entity to whom they > are addressed. If you are not the intended recipient or the individual > responsible for delivering the e-mail to the intended recipient, please > be advised that you have received this e-mail in error and that any > use, dissemination, forwarding, printing, or copying of this e-mail > is strictly prohibited. > > If you have received this e-mail in error, please immediately notify > the HealthPartners Support Center by telephone at (952) 967-6600. > You will be reimbursed for reasonable costs incurred in notifying us. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Fri Oct 1 17:35:42 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] NSH in ADVANCE for Medical Laboratory ProfessionalsEditorial References: Message-ID: <000801c4a806$fd65e4c0$5d30d445@domainnotset.invalid> I had problems with that link. This one worked for me. http://www.laboratorian.advanceweb.com/ Click on any of the words on the photo of Toronto. That should take you to the link page article. At the end of the article, there is a link to a list of all the NSH award winners. (Thank you vendors who generously sponsor these awards.) Hint to all histonetters - Deadline for next year's award will probably be around May 1, 2005. Start thinking now of applying for an award, or nominating someone. Nomination/application forms And there are lots of Histonetters who say they can't afford to go the a NSH Symposium - well - apply for a scholarship, and use the money to send yourself to a NSH meeting! If not for your own education, then to meet all the other histonetters! Application forms will be on line at the NSH web page soon. You DO have to be a NSH member in apply/win an award. For your convenience, the NSH membership page has links to both the NSH membership application form AND the NSH awards form pages. http://www.nsh.org/membership/ (Yes, I am shamelessly promoting NSH. And proud of it!) Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Luis Chiriboga" To: "Histonet" Sent: Friday, October 01, 2004 11:05 AM Subject: [Histonet] NSH in ADVANCE for Medical Laboratory ProfessionalsEditorial > Click on the link for synopsis of NSH meeting > > > http://www.laboratorian.advanceweb.com/common/Editorial/Editorial.aspx?CC=41 > 391 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pathrm35 <@t> adelphia.net Fri Oct 1 17:35:01 2004 From: pathrm35 <@t> adelphia.net (Ron Martin) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] salaries References: Message-ID: <019201c4a806$e4ad9db0$42e4aa45@Pathrm35> Two hurricanes in the past three weeks is enough for me. I spent all week moving into a new lab as our old one was destroyed by Jeanne. ----- Original Message ----- From: "Jes Strong - Source Medical Products" To: "Linda Blazek" ; ; Sent: Monday, September 27, 2004 2:25 PM Subject: RE: [Histonet] salaries > I hope Florida is still there. My heartfelt support for all of you who have > had to suffer the seemingly endless challenges you have had to face the past > couple of months. I know it may be of little condolence, but we all care and > pray for you. > > Jes Strong > Source Medical Products > Representatives for Statlab, DuraEdge, Bladex, Diagnostic BioSystems, RA > Lamb and other Quality Pathology Manufacturers > (847) 323-8373 (Cell) > (847) 735-9965 (Office) > (800) 442-3573 x250 (Voice Mail) > (508) 861-1575 (Fax) > jestrong@earthlink.net > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Linda > Blazek > Sent: Monday, September 27, 2004 2:08 PM > To: histonet@lists.utsouthwestern.edu; Laurie.Pereira@sdcounty.ca.gov > Subject: Re: [Histonet] salaries > > > > Brave woman, moving to Florida! > > Linda Blazek, HT (ASCP) > Department of Pathology > Children's Medical Center > 1 Children's Plaza > Dayton, Ohio 45404 > (937) 641-3358 > fax (937)641-5482 > blazekl@childrensdayton.org > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From RFail <@t> Charleston.net Fri Oct 1 20:47:22 2004 From: RFail <@t> Charleston.net (Rena) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Mounting media to covberslip from ethano Message-ID: <000001c4a821$c35b1550$d311a6a5@renad4yk9b8abe> Does anyone know of a mounting media that can be used to coverslip slides straight from ethanol and if so what are thw drawbacks. Rena Fail From beingmary53 <@t> ev1.net Fri Oct 1 22:51:44 2004 From: beingmary53 <@t> ev1.net (maryjohnson) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Alcoholic formalin Message-ID: <002301c4a833$23f713d0$3d49bbd0@ugly> Hi Dorothy here is the recipe for alcoholic formalin: Formaldehyde 37%-40%...............................200 ml Ethyl alcohol abs.........................................1300 ml Distilled water..............................................500 ml I use it in the place of 80% alcohol good luck Mary From ddeibler <@t> grandecom.net Sat Oct 2 07:18:14 2004 From: ddeibler <@t> grandecom.net (David Deibler) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] GI biosy embedding orientation Message-ID: <200410021218.i92CIKaZ008307@mx2.lsn.net> Our lab processes many small gi bxs on a daily basis. Our pathologists have been complaining about Incorrect orientation. We try to orient the specimens correctly but it seems to be hit or miss. Does anyone have suggestions or experience similar problems ??? David Deibler From ThisisAnn <@t> aol.com Sat Oct 2 08:06:34 2004 From: ThisisAnn <@t> aol.com (ThisisAnn@aol.com) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Technical Component Message-ID: <1d7.2c3f152a.2e9001da@aol.com> I would like to determine the cost of performing the Technical Component at my place of business. Is there a general "format" to follow (some kind of general guideline)? From gu.lang <@t> gmx.at Sat Oct 2 09:26:01 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Dewaxing in IHC Message-ID: <00b901c4a88b$be1cde60$eeeea8c0@server> Dear histonetters I would like to hear, what reagens is the most used in IHC for dewaxing the slides. We use xylen. And there is the opinion in our lab, that xylen is superior to other xylen-substitutes. Do you agree? For the HE-slides we use limonen, but for any delicate stain the boss demands xylen. thanks Gudrun From georgecole <@t> ev1.net Sat Oct 2 10:24:44 2004 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Susbstitute for barbitol based veronal acetate buffer Message-ID: <000001c4a893$f5648140$084dbad0@hppav> Histotechs!!! Lo and behold, I'm on GOOGLE.!!! I sent a message to the net answering an inquiry about replacing the barbitol buffer with something. As I had found Sigma 221 Alkaline Buffer to be better in every way, I sent a reply on the Histonet. Then, Mary North, the great gal who replaced me in the muscle and nerve lab here, informed me that the 221 buffer, once listed as 221 Alkaline buffer was now listed in the Sigma catalog as Alkaline Buffer A 9226. I sent a message to the histonet to that effect. Today, calling up GOOGLE, I entered 221 Alkaline buffer, and got a copy of my Histonet message about the change in the Sigma catalog number. YOIKS!!! I didn't know every message on the histonet would be copied for GOOGLE . I am now composing a three volume version of MY LIFE.--- Ah Hem!!!----, and I will send it to the net---then you may read it, I suppose, on GOOGLE under "Good ol' George" TA DAH!!! (Bows being taken.) From histophilhuff <@t> yahoo.com Sat Oct 2 14:34:11 2004 From: histophilhuff <@t> yahoo.com (Phillip Huff) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Dewaxing in IHC In-Reply-To: <00b901c4a88b$be1cde60$eeeea8c0@server> Message-ID: <20041002193411.66086.qmail@web50303.mail.yahoo.com> We are currently using EZ-dewax from bioGenex. It works great, is re-usable until the solution is saturated with wax and is very good for the tissues. http://www.biogenex.net/profile.php?pagename=autant1 Phil Gudrun Lang wrote: Dear histonetters I would like to hear, what reagens is the most used in IHC for dewaxing the slides. We use xylen. And there is the opinion in our lab, that xylen is superior to other xylen-substitutes. Do you agree? For the HE-slides we use limonen, but for any delicate stain the boss demands xylen. thanks Gudrun _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? New and Improved Yahoo! Mail - 100MB free storage! From rockbeki <@t> ufl.edu Sun Oct 3 09:22:05 2004 From: rockbeki <@t> ufl.edu (SMITH,REBEKAH FELICIA) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] enos Message-ID: <1292288368.1096813325767.JavaMail.osg@osgjas01.cns.ufl.edu> I'm doing IHC with Vegf currently on sheep placenta and over the summer I did IHC with eNOS also on sheep placenta. I just started the VEGF so I can't be much help there. On the eNOS, I can at least tell her what I did. Can you tell me more specifically what she wants to know? Rebekah Smith On Fri Oct 01 12:13:48 EDT 2004, Histology SLU wrote: > Hello All: > I have an investigator that is looking to do IHC using VegeF and > Enos antibodies on human lung tissue. Does anyone out there have > vendors, protocols, dilutions, or any information that you might > share with her. Thanks, in advance for your help. > Susan > > > --------------------------------- > Do you Yahoo!? > vote.yahoo.com - Register online to vote today! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" From rockbeki <@t> ufl.edu Sun Oct 3 09:32:49 2004 From: rockbeki <@t> ufl.edu (SMITH,REBEKAH FELICIA) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] IHC - weak signals Message-ID: <143466407.1096813969743.JavaMail.osg@osgjas02.cns.ufl.edu> I myself am more partial to AEC than to DAB (even though AEC can be real pain because its soluble in alcohol), because I find stuff more easily in bright red than in brown. Rebekah Smith On Fri Sep 24 10:19:25 EDT 2004, jason m wrote: > Does anyone have advice for amplifying weak signals from IHC? Is > there a detection system better than DAB which can make the > signal more noticable? > > _________________________________________________________________ > Designer Mail isn't just fun to send, it's fun to receive. Use > special stationery, fonts and colors. > http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines > Start enjoying all the benefits of MSN? Premium right now and > get the first two months FREE*. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" From rockbeki <@t> ufl.edu Sun Oct 3 09:43:57 2004 From: rockbeki <@t> ufl.edu (SMITH,REBEKAH FELICIA) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] question about inmuno Message-ID: <763813416.1096814637508.JavaMail.osg@osgjas02.cns.ufl.edu> I would think that it would depend highly on which chromagen you're using for your IHC. I have done IHC and H&E on the same slide, but it does make it a lot harder to see where your immunostaining is(especially since I was using brown DAB). My immunostaining was still there, btw, it was just really hard to see. Perhaps try to either get or make the DAB that turns black rather than brown (I think thats the nickel DAB) or use a chromagen thats a completely different color outside of the brownish, pinkish or bluish range. (I'm not sure what other chromogens there are since I've only used AEC and DAB but I know Vector makes some others.) Rebekah Smith On Thu Sep 23 12:39:35 EDT 2004, Jose Luis Palazon Fernandez wrote: > Dear List-Members > > > > First I want to thank the people who kindly answered to my > question about acid-resistant haematoxylin. I have a question > about inmunohistochemistry. I have to make a inmuno to detect > ATPase but then I would like to make another staining like PAS or > H&E. Could these "counterstainins" affect the inmuno reaction or > even cuould the inmuno response be lost or masked?. I want to > distinguish to kinds of cells, both of then are positive to PAS > but only one is positive to ATPase, I would like to show the two > in the same picture. Any help or comentary would be appreciated > > > > thanks in advance > > > > Jos? Luis > > > > > > > > > > El dia 20/09/2004 21:13 usted envio el siguiente mensaje: > >> Date: 20 de Septiembre de 2004 21:13:32 > >> From: "Gudrun Lang" > >> Subject: Re: [Histonet] hematoxylin > >> To: jluis.palazon@icman.csic.es > >> > >> I think the usual substitute here is Weigert's Hematoxylin. It >> stains the > >> nuclei black. We use it at the beginning of Trichrom-stains. > >> staining: 5 min (or more), differentiation with 0,5% HCl-Alk. >> (obtional), > >> blueing 5 min tapwater. > >> > >> working solution: > >> Gieson A + Gieson B 1:1 (stable for one week, older solutions >> stain rather > >> brownish than black) > >> > >> Gieson A: > >> 10 g hematoxylin + 1000 ml 96% Alk. (allow to stand for one week) > >> > >> Gieson B: > >> 40 ml 29% Ferric-chlorid > >> 980 ml Aqua dest > >> 10 ml 25% HCl > >> > >> greetings > >> Gudrun > >> > >> > >> ----- Original Message ----- > >> From: "Jose Luis Palazon Fernandez" > >> To: > >> Sent: Monday, September 20, 2004 8:25 PM > >> Subject: [Histonet] hematoxylin > >> > >> > >> Dear List-members > >> > >> greetings. I would like to know if there is a substitute for the >> "celestine > >> blue/hematoxylin" staining of nucleus that can resist a >> posterior acidic > >> counterstain. I need to apply a protocol that uses this nuclear >> staining > >> method but I dont have celestine blue and the company that >> suplies it says > >> that it would last 2 months to supply the stain > >> > >> Thanks in advance > >> > >> Jos? Luis > >> > >> _______________________________________________ > >> Histonet mailing list > >> Histonet@lists.utsouthwestern.edu > >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >> > > > > > > > > Universidad de Oriente-Isla Margarita-Venezuela > > actualmente en: Instituto de Ciencias Marinas de Andalucia > > Puerto Real, C?diz, Espa?a. > > email: jluis.palazon@icman.csic.es > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" From christopherjackson69 <@t> yahoo.ca Sun Oct 3 11:18:00 2004 From: christopherjackson69 <@t> yahoo.ca (Christopher Jackson) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Perfusion Fixation of a Mouse Heart Message-ID: <20041003161800.27218.qmail@web51703.mail.yahoo.com> Hi Folks, I am still looking for a protocol on how to perfusion fix a mouse heart and then embedd it in paraffin wax. I want to do as little physical damage as possible, because we want to study the morphology of the mouse's heart later. Any ideas? Thanks, Christopher Jackson Research Assistant Arrhthymia Research Group University of Ottawa Heart Institute 40 Ruskin Street Ottawa, ON K1Y 4W7 --------------------------------- Post your free ad now! Yahoo! Canada Personals From josephnerk <@t> hotmail.com Sun Oct 3 15:39:29 2004 From: josephnerk <@t> hotmail.com (Joseph Nerk) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Mailing list Message-ID: My island was hit by a hurrican so that is why all messages sent to me bounced as the internet system was down. _________________________________________________________________ Add photos to your e-mail with [1]MSN 8. Get 2 months FREE*. References 1. http://g.msn.com/8HMBEN/2746??PS=47575 From barbara.bublava <@t> meduniwien.ac.at Mon Oct 4 01:39:10 2004 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Mounting media to covberslip from ethano References: <000001c4a821$c35b1550$d311a6a5@renad4yk9b8abe> Message-ID: <000901c4a9dc$e1360690$1401a8c0@GERICHTS9XOZZ8> medite has a mounting medium called Mountex which can be used out of alcohol or isopropanol. I am testing it right now and can not see drawbacks. but i do not have expierience with longtime storage. Barbara Bublava ----- Original Message ----- From: "Rena" To: Sent: Saturday, October 02, 2004 3:47 AM Subject: [Histonet] Mounting media to covberslip from ethano > Does anyone know of a mounting media that can be used to coverslip > slides straight from ethanol and if so what are thw drawbacks. > > Rena Fail > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jluis.palazon <@t> icman.csic.es Mon Oct 4 02:40:37 2004 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] elastic fibers Message-ID: <20041004074037.0300F3127D5@perceval.uca.es> Dear List members I would like to know, in your experience, what is the best method to stain elatic fibers. Any help would be appreciated. thanks in advance Jos? Luis From rentonlf <@t> bru.wits.ac.za Mon Oct 4 04:54:29 2004 From: rentonlf <@t> bru.wits.ac.za (renton louise mrs) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] GI biosy embedding orientation Message-ID: <1096883669.936be3a0rentonlf@bru.wits.ac.za> For many years the lab where i worked used to get GI biopsies on pieces of filter paper, carboard etc where they had been placed immediately after biosy and before immersion in formalin (they seemed to stick down of their own accord).We then processed them still stuck down. This was supposed to assist with orientation at embedding as they were initially placed mucosa up, and could thus be rotated as needed. I have always wondered if this is/done anywhere else. louise -----Original Message----- From: "David Deibler" To: Date: Sat, 2 Oct 2004 07:18:14 -0500 Subject: [Histonet] GI biosy embedding orientation Our lab processes many small gi bxs on a daily basis. Our pathologists have been complaining about Incorrect orientation. We try to orient the specimens correctly but it seems to be hit or miss. Does anyone have suggestions or experience similar problems ??? David Deibler _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? From Julie.Sanders <@t> med.va.gov Mon Oct 4 06:16:10 2004 From: Julie.Sanders <@t> med.va.gov (Julie.Sanders@med.va.gov) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] IHC Equipment Message-ID: <457381D92B01BD44B21CF37CC02EBDFD28E96F@vhacinexc2.v10.med.va.gov> Ventana Benchmark. Julie Sanders,BA, HT(ASCP) Supervisor, Anatomic Pathology VAMC, Cincinnati, Oh. -- From Terry.Marshall <@t> rothgen.nhs.uk Mon Oct 4 06:25:56 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] GI biopsy embedding orientation Message-ID: The first point is that you must know *how* to embed them, which is at right angles to the curve or greatest curve if (as they usually are) they are curved in two planes. Aim for a section like this ...C. The second point is that you need to be able to see this, and therein lies the rub. You certainly need an aid (magnification and a good light). I have only seen this done in one place, Bristol Royal Infirmary, but they have the perfect solution. It is time consuming. The apparatus is a test tube rack of plastic insulin syringes, with the nozzle cut off, so that they are a simple straight tube with a piston. These are put in the rack, piston down, and a space left above, in the barrel, for the tissue. Gelatine is put in and the specimen oriented under a dissecting microscope. When cool, the piston is pushed to remove the small pellet which is embedded in paraffin in the usual way. | | | ~~~ | |-----| | | |_____| | || | | || | | || | | || | | || | | || | | || | | || | | || | _| || |_ || _||_ (Best I can do - these ascii artists must have infinite patience). Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: David Deibler [mailto:ddeibler@grandecom.net] Sent: 02 October 2004 13:18 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] GI biosy embedding orientation Our lab processes many small gi bxs on a daily basis. Our pathologists have been complaining about Incorrect orientation. We try to orient the specimens correctly but it seems to be hit or miss. Does anyone have suggestions or experience similar problems ??? David Deibler _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eenriquez52234 <@t> yahoo.com Mon Oct 4 06:46:38 2004 From: eenriquez52234 <@t> yahoo.com (esteban enriquez) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] IHC equipment In-Reply-To: Message-ID: <20041004114638.61864.qmail@web61107.mail.yahoo.com> Sorry to interrupt, but I heard that at NSH Vision Biosystems had their Bondmax at their booth. Not an impressive machine from the feedback as 1) you can't do ISH, 2) there's no standard existing predilutes, and 3) it's not a completely open system. How does the Nemesis stack up?? Thanks Esteban --- Joe Nocito wrote: > Janet, > I saw a new machine at the NSH from Biocare Medical > called the Nemesis. It > holds up to 84 slides, but the neat thing about it > is that you can have > programs running, and add more slides if you have to > just like the Tekmate > 1000. I miss my Tekmate because of the availability > of adding more slides > while the machine is running. Their numbe is > 800-799-9499 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On > Behalf Of > GUTIERREZ, JUAN > Sent: Friday, October 01, 2004 3:23 PM > To: Baez, Janet; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] IHC equipment > > > Ventana's Benchmarks. The best in the business. > > > Juan C. Gutierrez, HT(ASCP) > Histology Laboratory Supervisor > (210)704-2533 > > > -----Original Message----- > From: Baez, Janet [mailto:jbaez@interscopepath.com] > Sent: Tuesday, September 28, 2004 6:16 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] IHC equipment > > It's been 5 years since we did IHC in-house. We use > to run our IHC on > the old TechMate 1000. We're thinking of bringing > IHC back in-house. > Any feed back as to the new equipment available. > Any info will be > greatly appreciated. > Thanks. > > Janet E. Baez > Histology Manager > Interscope Pathology Medical Group > 21114 Vanowen St. > Canoga Park, Ca. > Tel. 818-992-7848 > Fax 818-992-6654 > _______________________________________________ > 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 > __________________________________ Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! http://promotions.yahoo.com/new_mail From eenriquez52234 <@t> yahoo.com Mon Oct 4 06:50:10 2004 From: eenriquez52234 <@t> yahoo.com (esteban enriquez) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Dewaxing in IHC In-Reply-To: <20041002193411.66086.qmail@web50303.mail.yahoo.com> Message-ID: <20041004115010.35402.qmail@web61102.mail.yahoo.com> Best thing I've heard used for dewaxing on certain systems - which can also pull double duty as a cleaner in tissue processors - is Isopar-L from Exxon-Mobil. Dirt cheap by the 5 gallon tin or 55 gallon drum. This is what Vision Biosystem use for dewax on their Bondmax and as a cleaner on their Peloris. Esteban --- Phillip Huff wrote: > We are currently using EZ-dewax from bioGenex. It > works great, is re-usable until the solution is > saturated with wax and is very good for the tissues. > > http://www.biogenex.net/profile.php?pagename=autant1 > > Phil > > Gudrun Lang wrote: > Dear histonetters > I would like to hear, what reagens is the most used > in IHC for dewaxing the slides. > We use xylen. And there is the opinion in our lab, > that xylen is superior to other xylen-substitutes. > Do you agree? > For the HE-slides we use limonen, but for any > delicate stain the boss demands xylen. > thanks > Gudrun > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > --------------------------------- > Do you Yahoo!? > New and Improved Yahoo! Mail - 100MB free storage! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________ Do you Yahoo!? New and Improved Yahoo! Mail - Send 10MB messages! http://promotions.yahoo.com/new_mail From Inga.Hansson <@t> neuro.uu.se Mon Oct 4 07:16:29 2004 From: Inga.Hansson <@t> neuro.uu.se (Inga Hansson) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Calbindin Message-ID: Hi everyone Has anyone used monoclonal anti-calbindin from Sigma on cryos? What fixation is best? PFA? Thanks in advance! Inga -- Inga Hansson dept. neuroscience, div. neurobiology PO Box 587 Biomedical Centre Husargatan 3 S-751 23 Uppsala SWEDEN phone:+46-18-4714384 fax: +46-18-559017 From asmith <@t> mail.barry.edu Mon Oct 4 07:54:19 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Mounting media to covberslip from ethano Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3BFB@exchsrv01.barrynet.barry.edu> Euparal, available from Carolina Biological Supply. The principal drawback is that some stains will fade during long-term exposure to alcohol. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rena Sent: Friday, October 01, 2004 9:47 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mounting media to covberslip from ethano Does anyone know of a mounting media that can be used to coverslip slides straight from ethanol and if so what are thw drawbacks. Rena Fail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From bruyntjes <@t> voeding.tno.nl Mon Oct 4 09:48:22 2004 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] (no subject) Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079D26@ntexch1.voeding.tno.nl> Hi everyone Is anyone of you aware of an antibody (mono- or polyclonal) that can be used to detect apoptosis in rat colon FFPE? J.P. Bruijntjes TNO Nutrition and Food Research Toxicology and Appllied Pharmacology The Netherlands This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From gcallis <@t> montana.edu Mon Oct 4 09:59:48 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Dewaxing in IHC In-Reply-To: <00b901c4a88b$be1cde60$eeeea8c0@server> References: <00b901c4a88b$be1cde60$eeeea8c0@server> Message-ID: <6.0.0.22.1.20041004085428.01b250e8@gemini.msu.montana.edu> We have used both Clearite 3 (xylene substitute) from Richard Allen, and xylene. A key thing is to do enough changes and longer times in these solvents to insure good paraffin removal, plus we keep them relatively fresh (rotation is done more frequently to insure first solvent does not contain lots of paraffin aka contaminated after many sections going through prior to IHC slides. We do 3 change, 5 minutes per change and either solvent for paraffin removal has worked well, no complaints from those doing IHC on paraffin sections. For all other staining, we use Clearite 3, 2 changes at 3 minutes each, but once again, keep reagents rotated. Limonene makes too many people sick in our lab, not allowed on the premises. At 08:26 AM 10/2/2004, you wrote: >Dear histonetters >I would like to hear, what reagens is the most used in IHC for dewaxing >the slides. >We use xylen. And there is the opinion in our lab, that xylen is superior >to other xylen-substitutes. Do you agree? >For the HE-slides we use limonen, but for any delicate stain the boss >demands xylen. >thanks >Gudrun >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Mon Oct 4 10:03:53 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] elastic fibers In-Reply-To: <20041004074037.0300F3127D5@perceval.uca.es> References: <20041004074037.0300F3127D5@perceval.uca.es> Message-ID: <6.0.0.22.1.20041004090251.01b43550@gemini.msu.montana.edu> Verhoeff's van Giesons for coarser fibers, for finer fibers AND coarse fibers, Weigerts Resorcin Fuchsin. At 01:40 AM 10/4/2004, you wrote: >Dear List members > >I would like to know, in your experience, what is the best method to stain >elatic fibers. Any help would be appreciated. > >thanks in advance > >Jos? Luis > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Sandra.Etheridge <@t> gems8.gov.bc.ca Mon Oct 4 10:09:15 2004 From: Sandra.Etheridge <@t> gems8.gov.bc.ca (Etheridge, Sandra AGF:EX) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Tissue Storage Solutions Message-ID: <424FAC08A8CDFE4BADD6497AE75B5E2F0E0A874B@atlas.gov.bc.ca> Hello, everyone, I have a couple issues I was hoping to get some feedback on: 1. I work in the provicial agricultural animal health lab, and our veterinary pathologists like to keep the original tissues from exotic animals, whales, sea lions, other interesting cases, indefinitely. We are currently transferring the tissues into 70% Ethanol, after about three months in 10% NBF, for long term storage. Does anyone else keep tissue past three months, and if so, what solution do you store them in? I have heard that formalin will keep hardening the tissues and make IHC next to impossible, and 70% will cause vacuolization of brain and may dehydrate over time. I have used both at different facilities I have worked in. Physiologic saline has been recommended, along with vacuum sealing the tissues into bags. Any comments or recommendations?? I'm not really sure why they want to hold on to the tissues, as we archive the blocks for future use. 2. My lab director was at a conference down in Texas recently and someone off-handedly mentioned that their recycled alcohols were messing up their IHC staining. I can't imagine why, and since we are considering the purchase of a recycler for our alochols and xylene, I thought I would see if anyone had similar problems. I know of one local hospital just using the gravity alcohol recycler filters, and their IHC is fine. Thanks for your time, everyone. As always, thanks for the feedback. Sandra Etheridge BC Ministry of Agriculture, Food & Fisheries Animal Health Center, Histology Abbotsford, BC Canada From Janet.Bonner <@t> FLHOSP.ORG Mon Oct 4 10:27:42 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] GI biopsy embedding orientation Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB408C@fh2k093.fhmis.net> Since many GI Bxs are taken with a spoon like device, embedding them on edge will give you correct orientation. Sometimes you can see the surface of the bx as a "white side", similar to a dermal Bx, and embed accordingly. Other GI Bxs will curl into a tube - embedding those on end with levels is the best bet. ( I used to have to orient gi biopsies for Electron Microscopy, so Histology was pretty straight forward). -Janet -----Original Message----- From: David Deibler [mailto:ddeibler@grandecom.net] Sent: Saturday, October 02, 2004 8:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] GI biosy embedding orientation Our lab processes many small gi bxs on a daily basis. Our pathologists have been complaining about Incorrect orientation. We try to orient the specimens correctly but it seems to be hit or miss. Does anyone have suggestions or experience similar problems ??? David Deibler _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From lizchlipala <@t> premierhistology.com Mon Oct 4 11:06:08 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] (no subject) In-Reply-To: <3B070848E7C2204F9DEB8BCFD767728001079D26@ntexch1.voeding.tno.nl> Message-ID: <000001c4aa2c$0f980340$76d48a80@AMY> J.P. Cleaved caspase 3 from Cell Signaling Technologies will work on FFPE specimens with high pH retreival. Your best bet for your positive control would be mesenteric lymph node. I have a protocol if you need one. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Histology Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 lizchlipala@premierhistology.com www.premierhistology.com Ship to Address: Premier Histology Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bruijntjes, J.P. Sent: Monday, October 04, 2004 7:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Hi everyone Is anyone of you aware of an antibody (mono- or polyclonal) that can be used to detect apoptosis in rat colon FFPE? J.P. Bruijntjes TNO Nutrition and Food Research Toxicology and Appllied Pharmacology The Netherlands This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carmen_loiselle <@t> hotmail.com Mon Oct 4 12:25:45 2004 From: carmen_loiselle <@t> hotmail.com (carmen loiselle) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Biodesine company Message-ID: Does anyone out there ever heard of that company "Biodesine", if so, would you be kind enough to send me the info how to contact them ? Thank you for your help, Carmen _________________________________________________________________ Des m?canismes de contr?le parental puissants permettent ? votre enfant de d?couvrir tout ce qu?Internet a ? offrir. http://join.msn.com/?pgmarket=fr-ca&page=features/parental&ST=1&xAPID=1983&DI=2043 Commencez d?s maintenant ? profiter de tous les avantages de MSN Premium et obtenez les deux premiers mois GRATUITS*. From shive003 <@t> umn.edu Mon Oct 4 13:10:55 2004 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Biodesine company References: Message-ID: <008f01c4aa3d$7e157940$78065486@vdl220FAC> Biodesign, Int. 60 Industrial Park Rd. Saco, ME 04072 USA Toll free US: 888-530-0140 Telephone: 207-283-6500 Fax: 207-283-4800 email: info@biodesign.com ----- Original Message ----- From: "carmen loiselle" To: Sent: Monday, October 04, 2004 12:25 PM Subject: [Histonet] Biodesine company > Does anyone out there ever heard of that company "Biodesine", if so, would > you be kind enough to send me the info how to contact them ? > > > Thank you for your help, > > Carmen > > _________________________________________________________________ > Des m?canismes de contr?le parental puissants permettent ? votre enfant de > d?couvrir tout ce qu'Internet a ? offrir. > http://join.msn.com/?pgmarket=fr-ca&page=features/parental&ST=1&xAPID=1983&DI=2043 > Commencez d?s maintenant ? profiter de tous les avantages de MSN Premium et > obtenez les deux premiers mois GRATUITS*. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Rcartun <@t> harthosp.org Mon Oct 4 13:16:02 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Biodesine company Message-ID: BioDesign International 60 Industrial Park Road Saco, ME 04072 (888) 530-0140 (207) 283-4800 Fax www.biodesign.com Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "carmen loiselle" 10/04/04 01:25PM >>> Does anyone out there ever heard of that company "Biodesine", if so, would you be kind enough to send me the info how to contact them ? Thank you for your help, Carmen _________________________________________________________________ Des m?canismes de contr?le parental puissants permettent ? votre enfant de d?couvrir tout ce qu'Internet a ? offrir. http://join.msn.com/?pgmarket=fr-ca&page=features/parental&ST=1&xAPID=1983&DI=2043 Commencez d?s maintenant ? profiter de tous les avantages de MSN Premium et obtenez les deux premiers mois GRATUITS*. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From billingconsultants <@t> yahoo.com Mon Oct 4 14:32:40 2004 From: billingconsultants <@t> yahoo.com (Caldwell) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Ventana Benchmark Attn: Vendors Message-ID: <20041004193240.93577.qmail@web54202.mail.yahoo.com> Hi Everyone, I am interested in obtaining quotes for Ventana's Benchmark for a client interested in doing HPV testing on Thin Prep paps. Any information you could provide would be greatly appreciated. Thank you in advance for your help. Lourena histonet-request@lists.utsouthwestern.edu wrote: Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Mailing list (Joseph Nerk) 2. Re: Mounting media to covberslip from ethano (Barbara Bublava) 3. elastic fibers (Jose Luis Palazon Fernandez) 4. Re: GI biosy embedding orientation (renton louise mrs) 5. IHC Equipment (Julie.Sanders@med.va.gov) 6. RE: GI biopsy embedding orientation (Marshall Terry Dr, Consultant Histopathologist) 7. RE: IHC equipment (esteban enriquez) 8. Re: Dewaxing in IHC (esteban enriquez) 9. Calbindin (Inga Hansson) 10. RE: Mounting media to covberslip from ethano (Smith, Allen) 11. (no subject) (Bruijntjes, J.P.) 12. Re: Dewaxing in IHC (Gayle Callis) 13. Re: elastic fibers (Gayle Callis) 14. Tissue Storage Solutions (Etheridge, Sandra AGF:EX) 15. RE: GI biopsy embedding orientation (Bonner, Janet) 16. RE: (no subject) (Elizabeth Chlipala) ---------------------------------------------------------------------- Message: 1 Date: Sun, 03 Oct 2004 15:39:29 -0500 From: "Joseph Nerk" Subject: [Histonet] Mailing list To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain My island was hit by a hurrican so that is why all messages sent to me bounced as the internet system was down. _________________________________________________________________ Add photos to your e-mail with [1]MSN 8. Get 2 months FREE*. References 1. http://g.msn.com/8HMBEN/2746??PS=47575 ------------------------------ Message: 2 Date: Mon, 4 Oct 2004 08:39:10 +0200 From: "Barbara Bublava" Subject: Re: [Histonet] Mounting media to covberslip from ethano To: "Rena" , Message-ID: <000901c4a9dc$e1360690$1401a8c0@GERICHTS9XOZZ8> Content-Type: text/plain; charset="iso-8859-1" medite has a mounting medium called Mountex which can be used out of alcohol or isopropanol. I am testing it right now and can not see drawbacks. but i do not have expierience with longtime storage. Barbara Bublava ----- Original Message ----- From: "Rena" To: Sent: Saturday, October 02, 2004 3:47 AM Subject: [Histonet] Mounting media to covberslip from ethano > Does anyone know of a mounting media that can be used to coverslip > slides straight from ethanol and if so what are thw drawbacks. > > Rena Fail > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Mon, 4 Oct 2004 09:40:37 +0200 (CEST) From: Jose Luis Palazon Fernandez Subject: [Histonet] elastic fibers To: histonet@lists.utsouthwestern.edu Message-ID: <20041004074037.0300F3127D5@perceval.uca.es> Content-Type: text/plain; charset="iso-8859-1" Dear List members I would like to know, in your experience, what is the best method to stain elatic fibers. Any help would be appreciated. thanks in advance José Luis ------------------------------ Message: 4 Date: Mon, 04 Oct 2004 11:54:29 +0200 From: "renton louise mrs" Subject: Re: [Histonet] GI biosy embedding orientation To: histonet@lists.utsouthwestern.edu Message-ID: <1096883669.936be3a0rentonlf@bru.wits.ac.za> Content-Type: text/plain; charset="UTF-8" For many years the lab where i worked used to get GI biopsies on pieces of filter paper, carboard etc where they had been placed immediately after biosy and before immersion in formalin (they seemed to stick down of their own accord).We then processed them still stuck down. This was supposed to assist with orientation at embedding as they were initially placed mucosa up, and could thus be rotated as needed. I have always wondered if this is/done anywhere else. louise -----Original Message----- From: "David Deibler" To: Date: Sat, 2 Oct 2004 07:18:14 -0500 Subject: [Histonet] GI biosy embedding orientation Our lab processes many small gi bxs on a daily basis. Our pathologists have been complaining about Incorrect orientation. We try to orient the specimens correctly but it seems to be hit or miss. Does anyone have suggestions or experience similar problems ??? David Deibler _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? ------------------------------ Message: 5 Date: Mon, 4 Oct 2004 06:16:10 -0500 From: Julie.Sanders@med.va.gov Subject: [Histonet] IHC Equipment To: histonet@lists.utsouthwestern.edu Message-ID: <457381D92B01BD44B21CF37CC02EBDFD28E96F@vhacinexc2.v10.med.va.gov> Content-Type: text/plain; charset="iso-8859-1" Ventana Benchmark. Julie Sanders,BA, HT(ASCP) Supervisor, Anatomic Pathology VAMC, Cincinnati, Oh. -- ------------------------------ Message: 6 Date: Mon, 4 Oct 2004 12:25:56 +0100 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] GI biopsy embedding orientation To: "David Deibler" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" The first point is that you must know *how* to embed them, which is at right angles to the curve or greatest curve if (as they usually are) they are curved in two planes. Aim for a section like this ...C. The second point is that you need to be able to see this, and therein lies the rub. You certainly need an aid (magnification and a good light). I have only seen this done in one place, Bristol Royal Infirmary, but they have the perfect solution. It is time consuming. The apparatus is a test tube rack of plastic insulin syringes, with the nozzle cut off, so that they are a simple straight tube with a piston. These are put in the rack, piston down, and a space left above, in the barrel, for the tissue. Gelatine is put in and the specimen oriented under a dissecting microscope. When cool, the piston is pushed to remove the small pellet which is embedded in paraffin in the usual way. | | | ~~~ | |-----| | | |_____| | || | | || | | || | | || | | || | | || | | || | | || | | || | _| || |_ || _||_ (Best I can do - these ascii artists must have infinite patience). Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: David Deibler [mailto:ddeibler@grandecom.net] Sent: 02 October 2004 13:18 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] GI biosy embedding orientation Our lab processes many small gi bxs on a daily basis. Our pathologists have been complaining about Incorrect orientation. We try to orient the specimens correctly but it seems to be hit or miss. Does anyone have suggestions or experience similar problems ??? David Deibler _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Mon, 4 Oct 2004 04:46:38 -0700 (PDT) From: esteban enriquez Subject: RE: [Histonet] IHC equipment To: Joe Nocito , "GUTIERREZ, JUAN" , "Baez, Janet" , histonet@lists.utsouthwestern.edu Message-ID: <20041004114638.61864.qmail@web61107.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Sorry to interrupt, but I heard that at NSH Vision Biosystems had their Bondmax at their booth. Not an impressive machine from the feedback as 1) you can't do ISH, 2) there's no standard existing predilutes, and 3) it's not a completely open system. How does the Nemesis stack up?? Thanks Esteban --- Joe Nocito wrote: > Janet, > I saw a new machine at the NSH from Biocare Medical > called the Nemesis. It > holds up to 84 slides, but the neat thing about it > is that you can have > programs running, and add more slides if you have to > just like the Tekmate > 1000. I miss my Tekmate because of the availability > of adding more slides > while the machine is running. Their numbe is > 800-799-9499 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On > Behalf Of > GUTIERREZ, JUAN > Sent: Friday, October 01, 2004 3:23 PM > To: Baez, Janet; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] IHC equipment > > > Ventana's Benchmarks. The best in the business. > > > Juan C. Gutierrez, HT(ASCP) > Histology Laboratory Supervisor > (210)704-2533 > > > -----Original Message----- > From: Baez, Janet [mailto:jbaez@interscopepath.com] > Sent: Tuesday, September 28, 2004 6:16 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] IHC equipment > > It's been 5 years since we did IHC in-house. We use > to run our IHC on > the old TechMate 1000. We're thinking of bringing > IHC back in-house. > Any feed back as to the new equipment available. > Any info will be > greatly appreciated. > Thanks. > > Janet E. Baez > Histology Manager > Interscope Pathology Medical Group > 21114 Vanowen St. > Canoga Park, Ca. > Tel. 818-992-7848 > Fax 818-992-6654 > _______________________________________________ > 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 > __________________________________ Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! http://promotions.yahoo.com/new_mail ------------------------------ Message: 8 Date: Mon, 4 Oct 2004 04:50:10 -0700 (PDT) From: esteban enriquez Subject: Re: [Histonet] Dewaxing in IHC To: Phillip Huff , Gudrun Lang , Histonetliste Message-ID: <20041004115010.35402.qmail@web61102.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Best thing I've heard used for dewaxing on certain systems - which can also pull double duty as a cleaner in tissue processors - is Isopar-L from Exxon-Mobil. Dirt cheap by the 5 gallon tin or 55 gallon drum. This is what Vision Biosystem use for dewax on their Bondmax and as a cleaner on their Peloris. Esteban --- Phillip Huff wrote: > We are currently using EZ-dewax from bioGenex. It > works great, is re-usable until the solution is > saturated with wax and is very good for the tissues. > > http://www.biogenex.net/profile.php?pagename=autant1 > > Phil > > Gudrun Lang wrote: > Dear histonetters > I would like to hear, what reagens is the most used > in IHC for dewaxing the slides. > We use xylen. And there is the opinion in our lab, > that xylen is superior to other xylen-substitutes. > Do you agree? > For the HE-slides we use limonen, but for any > delicate stain the boss demands xylen. > thanks > Gudrun > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > --------------------------------- > Do you Yahoo!? > New and Improved Yahoo! Mail - 100MB free storage! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________ Do you Yahoo!? New and Improved Yahoo! Mail - Send 10MB messages! http://promotions.yahoo.com/new_mail ------------------------------ Message: 9 Date: Mon, 4 Oct 2004 14:16:29 +0200 From: Inga Hansson Subject: [Histonet] Calbindin To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" Hi everyone Has anyone used monoclonal anti-calbindin from Sigma on cryos? What fixation is best? PFA? Thanks in advance! Inga -- Inga Hansson dept. neuroscience, div. neurobiology PO Box 587 Biomedical Centre Husargatan 3 S-751 23 Uppsala SWEDEN phone:+46-18-4714384 fax: +46-18-559017 ------------------------------ Message: 10 Date: Mon, 4 Oct 2004 08:54:19 -0400 From: "Smith, Allen" Subject: RE: [Histonet] Mounting media to covberslip from ethano To: "Rena" Cc: histonet@lists.utsouthwestern.edu Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3BFB@exchsrv01.barrynet.barry.edu> Content-Type: text/plain; charset="us-ascii" Euparal, available from Carolina Biological Supply. The principal drawback is that some stains will fade during long-term exposure to alcohol. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rena Sent: Friday, October 01, 2004 9:47 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mounting media to covberslip from ethano Does anyone know of a mounting media that can be used to coverslip slides straight from ethanol and if so what are thw drawbacks. Rena Fail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) ------------------------------ Message: 11 Date: Mon, 4 Oct 2004 16:48:22 +0200 From: "Bruijntjes, J.P." Subject: [Histonet] (no subject) To: Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079D26@ntexch1.voeding.tno.nl> Content-Type: text/plain; charset="us-ascii" Hi everyone Is anyone of you aware of an antibody (mono- or polyclonal) that can be used to detect apoptosis in rat colon FFPE? J.P. Bruijntjes TNO Nutrition and Food Research Toxicology and Appllied Pharmacology The Netherlands This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html ------------------------------ Message: 12 Date: Mon, 04 Oct 2004 08:59:48 -0600 From: Gayle Callis Subject: Re: [Histonet] Dewaxing in IHC To: "Gudrun Lang" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20041004085428.01b250e8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed We have used both Clearite 3 (xylene substitute) from Richard Allen, and xylene. A key thing is to do enough changes and longer times in these solvents to insure good paraffin removal, plus we keep them relatively fresh (rotation is done more frequently to insure first solvent does not contain lots of paraffin aka contaminated after many sections going through prior to IHC slides. We do 3 change, 5 minutes per change and either solvent for paraffin removal has worked well, no complaints from those doing IHC on paraffin sections. For all other staining, we use Clearite 3, 2 changes at 3 minutes each, but once again, keep reagents rotated. Limonene makes too many people sick in our lab, not allowed on the premises. At 08:26 AM 10/2/2004, you wrote: >Dear histonetters >I would like to hear, what reagens is the most used in IHC for dewaxing >the slides. >We use xylen. And there is the opinion in our lab, that xylen is superior >to other xylen-substitutes. Do you agree? >For the HE-slides we use limonen, but for any delicate stain the boss >demands xylen. >thanks >Gudrun >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 13 Date: Mon, 04 Oct 2004 09:03:53 -0600 From: Gayle Callis Subject: Re: [Histonet] elastic fibers To: Jose Luis Palazon Fernandez , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20041004090251.01b43550@gemini.msu.montana.edu> Content-Type: text/plain; charset="iso-8859-1"; format=flowed Verhoeff's van Giesons for coarser fibers, for finer fibers AND coarse fibers, Weigerts Resorcin Fuchsin. At 01:40 AM 10/4/2004, you wrote: >Dear List members > >I would like to know, in your experience, what is the best method to stain >elatic fibers. Any help would be appreciated. > >thanks in advance > >José Luis > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 14 Date: Mon, 4 Oct 2004 08:09:15 -0700 From: "Etheridge, Sandra AGF:EX" Subject: [Histonet] Tissue Storage Solutions To: " (histonet@lists.utsouthwestern.edu)" Message-ID: <424FAC08A8CDFE4BADD6497AE75B5E2F0E0A874B@atlas.gov.bc.ca> Content-Type: text/plain Hello, everyone, I have a couple issues I was hoping to get some feedback on: 1. I work in the provicial agricultural animal health lab, and our veterinary pathologists like to keep the original tissues from exotic animals, whales, sea lions, other interesting cases, indefinitely. We are currently transferring the tissues into 70% Ethanol, after about three months in 10% NBF, for long term storage. Does anyone else keep tissue past three months, and if so, what solution do you store them in? I have heard that formalin will keep hardening the tissues and make IHC next to impossible, and 70% will cause vacuolization of brain and may dehydrate over time. I have used both at different facilities I have worked in. Physiologic saline has been recommended, along with vacuum sealing the tissues into bags. Any comments or recommendations?? I'm not really sure why they want to hold on to the tissues, as we archive the blocks for future use. 2. My lab director was at a conference down in Texas recently and someone === message truncated === __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From MTitford <@t> aol.com Mon Oct 4 14:34:04 2004 From: MTitford <@t> aol.com (MTitford@aol.com) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Elastic time Message-ID: <4FDBBA8C.38FF9F86.00762DB1@aol.com> Jose Luis asks about elastic stains: Like Gayle in an earlier email, my personal preference is the Weigert Resorcin Fuchsin. It gives clearly stained elastic fibers without any background staining. Additionally, the stain used can be changed to give blue-green elastic fibers (Sheridan's) or red fibers (Weigert French modification) Mike Titford USA Pathology Mobile AL USA From tflore <@t> lsuhsc.edu Mon Oct 4 14:49:31 2004 From: tflore <@t> lsuhsc.edu (Flores, Teresa) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Hard of hearing, anyone? Message-ID: Dear Histonetters, perplexed with what I am working with and wondering if anyone else is having this trouble. With all the automated equipment in the labs, refrigerators, centrifuges, auto this and auto that, has anyone ever or routine performs a decibel reading in a histology lab? The reason I ask is that we have four ultramicrotones; a microwave, a building lab vent a hood, two counter top vents and two venting hoods which vent two processers, 3 refrigerators and a cryostat in a square footage of 20' x 25'. When someone walks in, I cannot hear a thing they are saying to me, I have to be right next to them and it is such a relief when I do walk out of the lab, so soothing to my ears, the quiet. Teresa From Shrttmprd <@t> aol.com Mon Oct 4 16:37:38 2004 From: Shrttmprd <@t> aol.com (Shrttmprd@aol.com) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Passing the HT exam Message-ID: <196.2fda3b48.2e931ca2@aol.com> I am accepting any and all advice to pass the HT exam. I have "The Theory and Practice od Histotechnology" book by Sheehan and Hrapchak. Is it true that whatever questions you don't answer on the exam WILL NOT be counted against you because somenone told me just to answer the questions I know. PLEASE HELP. Thank you, Melissa From POWELL_SA <@t> Mercer.edu Mon Oct 4 16:43:49 2004 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Hard of hearing, anyone? In-Reply-To: Message-ID: Hi Teresa, I have worked in the field for 42 years (yes I was 5 years old when I began), during that time in very noisy labs with all the equipment you mentioned running. Also I developed some major environmental allergies. All of this contributed to my now wearing 2 hearing aids. I have different deficits in each ear. Who can determine just what caused the loss, but I definitely had problems hearing conversation in the lab, even if they were standing next to me. I learned to read lips before I got my hearing aids, but that was no help if they were not looking straight at me. Who knows what I missed over the years, and how many co-workers I irritated asking them to repeat what they said. My insurance did not pay for the devices but did pay for the test to tell me I needed them. I had to sell my car at that time just to pay for them. I feel the noise level in the lab is as much a detriment to histotechs as is the repetitive motion injuries and the fumes we breath. That is another story in itself. Shirley -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Flores, Teresa Sent: Monday, October 04, 2004 2:50 PM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] Hard of hearing, anyone? Dear Histonetters, perplexed with what I am working with and wondering if anyone else is having this trouble. With all the automated equipment in the labs, refrigerators, centrifuges, auto this and auto that, has anyone ever or routine performs a decibel reading in a histology lab? The reason I ask is that we have four ultramicrotones; a microwave, a building lab vent a hood, two counter top vents and two venting hoods which vent two processers, 3 refrigerators and a cryostat in a square footage of 20' x 25'. When someone walks in, I cannot hear a thing they are saying to me, I have to be right next to them and it is such a relief when I do walk out of the lab, so soothing to my ears, the quiet. Teresa _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JSCHUMA1 <@t> FAIRVIEW.ORG Mon Oct 4 16:48:03 2004 From: JSCHUMA1 <@t> FAIRVIEW.ORG (JENNIFER SCHUMACHER) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Hard of hearing, anyone? Message-ID: I agree entirely, but have never done a decibel reading. Would be interested in other replys, as my techs have recently been asking the same question. So much automation can hurt the ears:-) Jen >>> "Flores, Teresa" 10/04/04 02:49PM >>> Dear Histonetters, perplexed with what I am working with and wondering if anyone else is having this trouble. With all the automated equipment in the labs, refrigerators, centrifuges, auto this and auto that, has anyone ever or routine performs a decibel reading in a histology lab? The reason I ask is that we have four ultramicrotones; a microwave, a building lab vent a hood, two counter top vents and two venting hoods which vent two processers, 3 refrigerators and a cryostat in a square footage of 20' x 25'. When someone walks in, I cannot hear a thing they are saying to me, I have to be right next to them and it is such a relief when I do walk out of the lab, so soothing to my ears, the quiet. Teresa _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted in this e-mail is intended only for the person or entity to which it is addressed and may contain confidential and/or privileged material, including “protected health information.” If you are not the intended recipient, you are hereby notified that any review, retransmission, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. <<<>>> From JMacDonald <@t> mtsac.edu Mon Oct 4 16:58:57 2004 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Hard of hearing, anyone? In-Reply-To: Message-ID: I believe OSHA has a standard regarding sound and hearing testing. "JENNIFER SCHUMACHER" Sent by: histonet-bounces@lists.utsouthwestern.edu 10/04/2004 02:48 PM To , cc Subject Re: [Histonet] Hard of hearing, anyone? I agree entirely, but have never done a decibel reading. Would be interested in other replys, as my techs have recently been asking the same question. So much automation can hurt the ears:-) Jen >>> "Flores, Teresa" 10/04/04 02:49PM >>> Dear Histonetters, perplexed with what I am working with and wondering if anyone else is having this trouble. With all the automated equipment in the labs, refrigerators, centrifuges, auto this and auto that, has anyone ever or routine performs a decibel reading in a histology lab? The reason I ask is that we have four ultramicrotones; a microwave, a building lab vent a hood, two counter top vents and two venting hoods which vent two processers, 3 refrigerators and a cryostat in a square footage of 20' x 25'. When someone walks in, I cannot hear a thing they are saying to me, I have to be right next to them and it is such a relief when I do walk out of the lab, so soothing to my ears, the quiet. Teresa _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted in this e-mail is intended only for the person or entity to which it is addressed and may contain confidential and/or privileged material, including ?protected health information.? If you are not the intended recipient, you are hereby notified that any review, retransmission, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. <<<>>> _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JMacDonald <@t> mtsac.edu Mon Oct 4 17:10:53 2004 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Passing the HT exam In-Reply-To: <196.2fda3b48.2e931ca2@aol.com> Message-ID: Points are awarded for correct responses. The value of each question is based upon the level of difficulty. The more difficult the question, the more points awarded. While you are not penalized for answering a question incorrectly, you are not accumulating points. You must achieve a score of 400 or greater to pass. There are 100 questions. 10-25% of the questions will relate to fixation. 10-14% will relate to processing and embedding. 10-14% will relate to microtomy. 40-50% will relate to staining. 10-15% will relate to laboratory operations (safety, instrumentation, lab math, regulations) Jennifer MacDonald Shrttmprd@aol.com Sent by: histonet-bounces@lists.utsouthwestern.edu 10/04/2004 02:37 PM To histonet@lists.utsouthwestern.edu cc Subject [Histonet] Passing the HT exam I am accepting any and all advice to pass the HT exam. I have "The Theory and Practice od Histotechnology" book by Sheehan and Hrapchak. Is it true that whatever questions you don't answer on the exam WILL NOT be counted against you because somenone told me just to answer the questions I know. PLEASE HELP. Thank you, Melissa _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Mon Oct 4 19:37:24 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] elastic fibers References: <20041004074037.0300F3127D5@perceval.uca.es> <6.0.0.22.1.20041004090251.01b43550@gemini.msu.montana.edu> Message-ID: <005b01c4aa73$7c9de4e0$7fe0d445@domainnotset.invalid> Another one for the fine and coarse fibers that seems to be gaining popularity is the Miller stain. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Gayle Callis" To: "Jose Luis Palazon Fernandez" ; Sent: Monday, October 04, 2004 11:03 AM Subject: Re: [Histonet] elastic fibers Verhoeff's van Giesons for coarser fibers, for finer fibers AND coarse fibers, Weigerts Resorcin Fuchsin. At 01:40 AM 10/4/2004, you wrote: >Dear List members > >I would like to know, in your experience, what is the best method to stain >elatic fibers. Any help would be appreciated. > >thanks in advance > >Jos? Luis > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From info <@t> ihcworld.com Mon Oct 4 21:03:49 2004 From: info <@t> ihcworld.com (IHC World) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] IHC World Image Gallery Launched Message-ID: We have recently launched a new site - IHC World Image Gallery (http://www.ihcworld.com/image_gallery.htm) The aim is to enhance communication and discussion among researchers in histology, histopathology and immunohistochemistry community. Users (upon registration) can create your own album, upload images, view and post comments. You can also search images of interest within the gallery. Everyone is welcome to use this image gallery as a tool to enhance communication and discussion when it is needed. Richard IHC World From Andres.Kulla <@t> kliinikum.ee Tue Oct 5 01:10:38 2004 From: Andres.Kulla <@t> kliinikum.ee (Andres Kulla) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] tissue processors Message-ID: <687732B2C3C2A845A03EAD8B00640C2D01DEB7@viplala.kliinikum.ee> To the experts in automated tissue processing, Could anybody share his/her experience with TBS (Triangle Biomedical Sciences) ATP1 tissue processor. Positive vs negative aspects in comparison with other devices eg Sakuras Tissue Tek VIP 500. With best regards from Andres Kulla, Estonia From ctsblack <@t> capeheart.uct.ac.za Tue Oct 5 02:46:48 2004 From: ctsblack <@t> capeheart.uct.ac.za (Melanie Black) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Abcam CD 3 antibody. Message-ID: Hi There I am setting up ABCAMS CD 3 antibody. Would anybody have the optimun dilution for this antibody, and the antigen retrieval? I am currenly trying EDTA buffer pH 8.0 for 3x5 mins in the microwave, and have set up dilutions from 1;20/1:50 and 1:100. Being fluorescent, I am not sure if I am detecting all the lymphocytes. I am using human control tonsil. This is a RAT - Monoclonal, and I am therefore using the recommended Anti Rat (Cy 5 linked) secondary anitbody. Many Thanks Melanie. -- Cardiovascular Research Unit Div. of Cardiothoracic Surgery Chris Barnard Building University of Cape Town Anzio Road Observatory 7925 Republic of South Africa Tel +27 21 406-6589 Cel +27 82 469-3352 Fax +27 21 448-5935 From carl.hobbs <@t> kcl.ac.uk Tue Oct 5 03:38:49 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] re Calbindin Message-ID: <001d01c4aab6$bcc62ad0$e8345c9f@Carlos> Haven't used it on frozens but it works beautifully on Formalin- fixed p.wax sections, after AR From c.m.vanderloos <@t> amc.uva.nl Tue Oct 5 04:40:22 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] RE: Permanent Red Message-ID: <5e1a55fe95.5fe955e1a5@amc.uva.nl> Hi Patty, Indeed Dako's Permanent Red is a very nice chromogen that has a really crisp localization. However, we also found that it dissolves a bit in alcohol. The alcohol we used was not recycled and completely clean. According to the data sheet Permanent Red is not really designed to be solved in organic-based mountants. We let the slide dry completely on a hot plate (after washing with distilled water) and use VectaMount (H-5000). This is a non-aqueous mountant that works perfectly well on paraffin slides. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Patti Loykasek Date Fri, 01 Oct 2004 07:38:18 -0700 To histonet Cc Bret Cook Subject [Histonet] Permanent Red Just a quick question on alternative chromogens. Has anyone had a problem with Dako?s Permanent Red coming out in recycled alcohols? This is a nice alk. Phos. Chromogen, and I really like it. Occasionally though, it comes out in the alcohols in our counterstaining/dehydrating set up ? not every time. We do use recycled alcohols in this set-up. If we switch the recycled out for all fresh, we don?t have this problem. Just wondering. Patti Loykasek PhenoPath Laboratories Seattle, WA From joycejudge259 <@t> hotmail.com Tue Oct 5 06:15:54 2004 From: joycejudge259 <@t> hotmail.com (joyce judge) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] mouse paws Message-ID: I would like too know if people that are processing mouse paws deglove or process with the skin on. Joyce Judge Visen Medical jjudge@visenmedical.com _________________________________________________________________ Is your PC infected? Get a FREE online computer virus scan from McAfee® Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From eenriquez52234 <@t> yahoo.com Tue Oct 5 08:56:42 2004 From: eenriquez52234 <@t> yahoo.com (esteban enriquez) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Ventana Benchmark Attn: Vendors In-Reply-To: <20041004193240.93577.qmail@web54202.mail.yahoo.com> Message-ID: <20041005135643.29749.qmail@web61105.mail.yahoo.com> Good choice! Verntana may be better than Vision Biosystem Bond system, or Biogenex or Biocare or Dako. From NSH, many shortcoming with no fix in near future. Ask anyone in Histo , other Companies doesn't have science needed to compete in open form. Esteban Caldwell wrote: Hi Everyone, I am interested in obtaining quotes for Ventana's Benchmark for a client interested in doing HPV testing on Thin Prep paps. Any information you could provide would be greatly appreciated. Thank you in advance for your help. Lourena histonet-request@lists.utsouthwestern.edu wrote: Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Mailing list (Joseph Nerk) 2. Re: Mounting media to covberslip from ethano (Barbara Bublava) 3. elastic fibers (Jose Luis Palazon Fernandez) 4. Re: GI biosy embedding orientation (renton louise mrs) 5. IHC Equipment (Julie.Sanders@med.va.gov) 6. RE: GI biopsy embedding orientation (Marshall Terry Dr, Consultant Histopathologist) 7. RE: IHC equipment (esteban enriquez) 8. Re: Dewaxing in IHC (esteban enriquez) 9. Calbindin (Inga Hansson) 10. RE: Mounting media to covberslip from ethano (Smith, Allen) 11. (no subject) (Bruijntjes, J.P.) 12. Re: Dewaxing in IHC (Gayle Callis) 13. Re: elastic fibers (Gayle Callis) 14. Tissue Storage Solutions (Etheridge, Sandra AGF:EX) 15. RE: GI biopsy embedding orientation (Bonner, Janet) 16. RE: (no subject) (Elizabeth Chlipala) ---------------------------------------------------------------------- Message: 1 Date: Sun, 03 Oct 2004 15:39:29 -0500 From: "Joseph Nerk" Subject: [Histonet] Mailing list To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain My island was hit by a hurrican so that is why all messages sent to me bounced as the internet system was down. _________________________________________________________________ Add photos to your e-mail with [1]MSN 8. Get 2 months FREE*. References 1. http://g.msn.com/8HMBEN/2746??PS=47575 ------------------------------ Message: 2 Date: Mon, 4 Oct 2004 08:39:10 +0200 From: "Barbara Bublava" Subject: Re: [Histonet] Mounting media to covberslip from ethano To: "Rena" , Message-ID: <000901c4a9dc$e1360690$1401a8c0@GERICHTS9XOZZ8> Content-Type: text/plain; charset="iso-8859-1" medite has a mounting medium called Mountex which can be used out of alcohol or isopropanol. I am testing it right now and can not see drawbacks. but i do not have expierience with longtime storage. Barbara Bublava ----- Original Message ----- From: "Rena" To: Sent: Saturday, October 02, 2004 3:47 AM Subject: [Histonet] Mounting media to covberslip from ethano > Does anyone know of a mounting media that can be used to coverslip > slides straight from ethanol and if so what are thw drawbacks. > > Rena Fail > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Mon, 4 Oct 2004 09:40:37 +0200 (CEST) From: Jose Luis Palazon Fernandez Subject: [Histonet] elastic fibers To: histonet@lists.utsouthwestern.edu Message-ID: <20041004074037.0300F3127D5@perceval.uca.es> Content-Type: text/plain; charset="iso-8859-1" Dear List members I would like to know, in your experience, what is the best method to stain elatic fibers. Any help would be appreciated. thanks in advance José Luis ------------------------------ Message: 4 Date: Mon, 04 Oct 2004 11:54:29 +0200 From: "renton louise mrs" Subject: Re: [Histonet] GI biosy embedding orientation To: histonet@lists.utsouthwestern.edu Message-ID: <1096883669.936be3a0rentonlf@bru.wits.ac.za> Content-Type: text/plain; charset="UTF-8" For many years the lab where i worked used to get GI biopsies on pieces of filter paper, carboard etc where they had been placed immediately after biosy and before immersion in formalin (they seemed to stick down of their own accord).We then processed them still stuck down. This was supposed to assist with orientation at embedding as they were initially placed mucosa up, and could thus be rotated as needed. I have always wondered if this is/done anywhere else. louise -----Original Message----- From: "David Deibler" To: Date: Sat, 2 Oct 2004 07:18:14 -0500 Subject: [Histonet] GI biosy embedding orientation Our lab processes many small gi bxs on a daily basis. Our pathologists have been complaining about Incorrect orientation. We try to orient the specimens correctly but it seems to be hit or miss. Does anyone have suggestions or experience similar problems ??? David Deibler _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? ------------------------------ Message: 5 Date: Mon, 4 Oct 2004 06:16:10 -0500 From: Julie.Sanders@med.va.gov Subject: [Histonet] IHC Equipment To: histonet@lists.utsouthwestern.edu Message-ID: <457381D92B01BD44B21CF37CC02EBDFD28E96F@vhacinexc2.v10.med.va.gov> Content-Type: text/plain; charset="iso-8859-1" Ventana Benchmark. Julie Sanders,BA, HT(ASCP) Supervisor, Anatomic Pathology VAMC, Cincinnati, Oh. -- ------------------------------ Message: 6 Date: Mon, 4 Oct 2004 12:25:56 +0100 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] GI biopsy embedding orientation To: "David Deibler" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" The first point is that you must know *how* to embed them, which is at right angles to the curve or greatest curve if (as they usually are) they are curved in two planes. Aim for a section like this ...C. The second point is that you need to be able to see this, and therein lies the rub. You certainly need an aid (magnification and a good light). I have only seen this done in one place, Bristol Royal Infirmary, but they have the perfect solution. It is time consuming. The apparatus is a test tube rack of plastic insulin syringes, with the nozzle cut off, so that they are a simple straight tube with a piston. These are put in the rack, piston down, and a space left above, in the barrel, for the tissue. Gelatine is put in and the specimen oriented under a dissecting microscope. When cool, the piston is pushed to remove the small pellet which is embedded in paraffin in the usual way. | | | ~~~ | |-----| | | |_____| | || | | || | | || | | || | | || | | || | | || | | || | | || | _| || |_ || _||_ (Best I can do - these ascii artists must have infinite patience). Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: David Deibler [mailto:ddeibler@grandecom.net] Sent: 02 October 2004 13:18 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] GI biosy embedding orientation Our lab processes many small gi bxs on a daily basis. Our pathologists have been complaining about Incorrect orientation. We try to orient the specimens correctly but it seems to be hit or miss. Does anyone have suggestions or experience similar problems ??? David Deibler _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Mon, 4 Oct 2004 04:46:38 -0700 (PDT) From: esteban enriquez Subject: RE: [Histonet] IHC equipment To: Joe Nocito , "GUTIERREZ, JUAN" , "Baez, Janet" , histonet@lists.utsouthwestern.edu Message-ID: <20041004114638.61864.qmail@web61107.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Sorry to interrupt, but I heard that at NSH Vision Biosystems had their Bondmax at their booth. Not an impressive machine from the feedback as 1) you can't do ISH, 2) there's no standard existing predilutes, and 3) it's not a completely open system. How does the Nemesis stack up?? Thanks Esteban --- Joe Nocito wrote: > Janet, > I saw a new machine at the NSH from Biocare Medical > called the Nemesis. It > holds up to 84 slides, but the neat thing about it > is that you can have > programs running, and add more slides if you have to > just like the Tekmate > 1000. I miss my Tekmate because of the availability > of adding more slides > while the machine is running. Their numbe is > 800-799-9499 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On > Behalf Of > GUTIERREZ, JUAN > Sent: Friday, October 01, 2004 3:23 PM > To: Baez, Janet; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] IHC equipment > > > Ventana's Benchmarks. The best in the business. > > > Juan C. Gutierrez, HT(ASCP) > Histology Laboratory Supervisor > (210)704-2533 > > > -----Original Message----- > From: Baez, Janet [mailto:jbaez@interscopepath.com] > Sent: Tuesday, September 28, 2004 6:16 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] IHC equipment > > It's been 5 years since we did IHC in-house. We use > to run our IHC on > the old TechMate 1000. We're thinking of bringing > IHC back in-house. > Any feed back as to the new equipment available. > Any info will be > greatly appreciated. > Thanks. > > Janet E. Baez > Histology Manager > Interscope Pathology Medical Group > 21114 Vanowen St. > Canoga Park, Ca. > Tel. 818-992-7848 > Fax 818-992-6654 > _______________________________________________ > 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 > __________________________________ Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! http://promotions.yahoo.com/new_mail ------------------------------ Message: 8 Date: Mon, 4 Oct 2004 04:50:10 -0700 (PDT) From: esteban enriquez Subject: Re: [Histonet] Dewaxing in IHC To: Phillip Huff , Gudrun Lang , Histonetliste Message-ID: <20041004115010.35402.qmail@web61102.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Best thing I've heard used for dewaxing on certain systems - which can also pull double duty as a cleaner in tissue processors - is Isopar-L from Exxon-Mobil. Dirt cheap by the 5 gallon tin or 55 gallon drum. This is what Vision Biosystem use for dewax on their Bondmax and as a cleaner on their Peloris. Esteban --- Phillip Huff wrote: > We are currently using EZ-dewax from bioGenex. It > works great, is re-usable until the solution is > saturated with wax and is very good for the tissues. > > http://www.biogenex.net/profile.php?pagename=autant1 > > Phil > > Gudrun Lang wrote: > Dear histonetters > I would like to hear, what reagens is the most used > in IHC for dewaxing the slides. > We use xylen. And there is the opinion in our lab, > that xylen is superior to other xylen-substitutes. > Do you agree? > For the HE-slides we use limonen, but for any > delicate stain the boss demands xylen. > thanks > Gudrun > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > --------------------------------- > Do you Yahoo!? > New and Improved Yahoo! Mail - 100MB free storage! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________ Do you Yahoo!? New and Improved Yahoo! Mail - Send 10MB messages! http://promotions.yahoo.com/new_mail ------------------------------ Message: 9 Date: Mon, 4 Oct 2004 14:16:29 +0200 From: Inga Hansson Subject: [Histonet] Calbindin To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" Hi everyone Has anyone used monoclonal anti-calbindin from Sigma on cryos? What fixation is best? PFA? Thanks in advance! Inga -- Inga Hansson dept. neuroscience, div. neurobiology PO Box 587 Biomedical Centre Husargatan 3 S-751 23 Uppsala SWEDEN phone:+46-18-4714384 fax: +46-18-559017 ------------------------------ Message: 10 Date: Mon, 4 Oct 2004 08:54:19 -0400 From: "Smith, Allen" Subject: RE: [Histonet] Mounting media to covberslip from ethano To: "Rena" Cc: histonet@lists.utsouthwestern.edu Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3BFB@exchsrv01.barrynet.barry.edu> Content-Type: text/plain; charset="us-ascii" Euparal, available from Carolina Biological Supply. The principal drawback is that some stains will fade during long-term exposure to alcohol. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rena Sent: Friday, October 01, 2004 9:47 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mounting media to covberslip from ethano Does anyone know of a mounting media that can be used to coverslip slides straight from ethanol and if so what are thw drawbacks. Rena Fail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) ------------------------------ Message: 11 Date: Mon, 4 Oct 2004 16:48:22 +0200 From: "Bruijntjes, J.P." Subject: [Histonet] (no subject) To: Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079D26@ntexch1.voeding.tno.nl> Content-Type: text/plain; charset="us-ascii" Hi everyone Is anyone of you aware of an antibody (mono- or polyclonal) that can be used to detect apoptosis in rat colon FFPE? J.P. Bruijntjes TNO Nutrition and Food Research Toxicology and Appllied Pharmacology The Netherlands This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html ------------------------------ Message: 12 Date: Mon, 04 Oct 2004 08:59:48 -0600 From: Gayle Callis Subject: Re: [Histonet] Dewaxing in IHC To: "Gudrun Lang" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20041004085428.01b250e8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed We have used both Clearite 3 (xylene substitute) from Richard Allen, and xylene. A key thing is to do enough changes and longer times in these solvents to insure good paraffin removal, plus we keep them relatively fresh (rotation is done more frequently to insure first solvent does not contain lots of paraffin aka contaminated after many sections going through prior to IHC slides. We do 3 change, 5 minutes per change and either solvent for paraffin removal has worked well, no complaints from those doing IHC on paraffin sections. For all other staining, we use Clearite 3, 2 changes at 3 minutes each, but once again, keep reagents rotated. Limonene makes too many people sick in our lab, not allowed on the premises. At 08:26 AM 10/2/2004, you wrote: >Dear histonetters >I would like to hear, what reagens is the most used in IHC for dewaxing >the slides. >We use xylen. And there is the opinion in our lab, that xylen is superior >to other xylen-substitutes. Do you agree? >For the HE-slides we use limonen, but for any delicate stain the boss >demands xylen. >thanks >Gudrun >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 13 Date: Mon, 04 Oct 2004 09:03:53 -0600 From: Gayle Callis Subject: Re: [Histonet] elastic fibers To: Jose Luis Palazon Fernandez , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20041004090251.01b43550@gemini.msu.montana.edu> Content-Type: text/plain; charset="iso-8859-1"; format=flowed Verhoeff's van Giesons for coarser fibers, for finer fibers AND coarse fibers, Weigerts Resorcin Fuchsin. At 01:40 AM 10/4/2004, you wrote: >Dear List members > >I would like to know, in your experience, what is the best method to stain >elatic fibers. Any help would be appreciated. > >thanks in advance > >José Luis > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 14 Date: Mon, 4 Oct 2004 08:09:15 -0700 From: "Etheridge, Sandra AGF:EX" Subject: [Histonet] Tissue Storage Solutions To: " (histonet@lists.utsouthwestern.edu)" Message-ID: <424FAC08A8CDFE4BADD6497AE75B5E2F0E0A874B@atlas.gov.bc.ca> Content-Type: text/plain Hello, everyone, I have a couple issues I was hoping to get some feedback on: 1. I work in the provicial agricultural animal health lab, and our veterinary pathologists like to keep the original tissues from exotic animals, whales, sea lions, other interesting cases, indefinitely. We are currently transferring the tissues into 70% Ethanol, after about three months in 10% NBF, for long term storage. Does anyone else keep tissue past three months, and if so, what solution do you store them in? I have heard that formalin will keep hardening the tissues and make IHC next to impossible, and 70% will cause vacuolization of brain and may dehydrate over time. I have used both at different facilities I have worked in. Physiologic saline has been recommended, along with vacuum sealing the tissues into bags. Any comments or recommendations?? I'm not really sure why they want to hold on to the tissues, as we archive the blocks for future use. 2. My lab director was at a conference down in Texas recently and someone === message truncated === __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From gcallis <@t> montana.edu Tue Oct 5 09:30:42 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] mouse paws In-Reply-To: References: Message-ID: <6.0.0.22.1.20041005082945.01b0f408@gemini.msu.montana.edu> No we leave paws intact. However, mouse paws are extremely dense even though tiny, and after decalcification, we do a longer processing schedule. At 05:15 AM 10/5/2004, you wrote: >I would like too know if people that are processing mouse paws deglove or >process with the skin on. > >Joyce Judge >Visen Medical >jjudge@visenmedical.com > >_________________________________________________________________ >Is your PC infected? Get a FREE online computer virus scan from McAfee? >Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From contact <@t> excaliburpathology.com Tue Oct 5 09:41:44 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Tissue storage solutions Message-ID: <20041005144144.42603.qmail@web50306.mail.yahoo.com> I have always stored the wet tissue, not processed for paraffin sections from the whole eye specimens I receive, in 70% EtOH. I keep them for a minimum of 3 years, but if you change the 70% every couple of years, you can keep the specimens indefinantly. Paula Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From pedro.louro <@t> spcorp.com Tue Oct 5 09:48:28 2004 From: pedro.louro <@t> spcorp.com (Louro, Pedro) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] DAB-colors... Message-ID: <4508920F80C0D411B90200508BF9A9F4062B4727@LAFMSG30.us.schp.com> Hello all, When at NSH -Toronto..I heard people talking about using DAB in different colors. Can someone tell who sells this product. Thanks Pedro ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From contact <@t> excaliburpathology.com Tue Oct 5 09:49:43 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Mouse paws Message-ID: <20041005144943.46312.qmail@web50306.mail.yahoo.com> I did some mouse paws for a researcher interested in the joints for arthritis and taking the skin off made sectioning much easier. The hair on the skin will scratch your knife quickly. So if it is not what they need to see, take if off and save the edge of your blades. Paula Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From histology.bc <@t> shaw.ca Tue Oct 5 11:17:18 2004 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Mounting media to covberslip from ethano In-Reply-To: <000001c4a821$c35b1550$d311a6a5@renad4yk9b8abe> References: <000001c4a821$c35b1550$d311a6a5@renad4yk9b8abe> Message-ID: <4162C90E.6020303@shaw.ca> I have never encountered a mounting medium that uses alcohol as the solvent. If one exists, two disadvantages spring to mind immediately. One, the refractive index of the medium would be low, so the clarity and brilliance associated with conventional, xylene-based media would be absent. Two, a large proportion of counterstains (eosin, van Gieson, neutral red, methylene blue, etc) are alcohol -soluble and would "bleed" into the medium. Paul From jkiernan <@t> uwo.ca Tue Oct 5 11:05:06 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Message for Mark Ray References: <000001c4a68d$03c24780$c211a6a5@renad4yk9b8abe> <415B7D83.64C14C0A@uwo.ca> <001c01c4a6d1$7acdf470$eeeea8c0@server> <415DAE00.E56C7AAE@uwo.ca> <41629370.60703@mindspring.com> Message-ID: <4162C632.E5C90763@uwo.ca> Dear Mark, Our mail server says your email address darkdaym@mindspring.com does not exist, so I'm sending the message to Histonet instead, with your name in the subject line. Yes, safranine has an e and so does rhodamine. Acid fuchsine gets an e because although it's an anionic dye it's derived from an organic base by addition of sulfonic acid groups. If you're interested I can send you a reprint of a 2001 paper in Biotech. Histochem. on dye nomenclature; just let me know where to send it to. It's similar to Ch. 3 in Conn's Biol. St. Conn's 10th edn uses US spellings (despite a Brit publisher) but in dye names this differs only slightly from English spelling. The only differences I can think of offhand are having an f instead of ph in parts of words derived from sulfur, and e rather than ae in hematoxylin and hematein. We hope suppliers will adopt Conn's as a standard for names - not only spellings but also calling the dyes what they really are. Ethyl green is an example of one that's probably been wrongly labeled for 30+ years despite the fact that it's better for its job than methyl green. Best regards, John. ________________________________________ Mark Ray wrote: > John, > > Help me out here. Is the terminal e required in safranin_e_? It's an > amine, right? I've noticed that JT Baker has added the e on its MSD > Sheet but not in its catalog, as yet. Are the spellings in the new > Conn to be considered universally authoritative or at least correct in > US dialect? I think I may have a lot of catalog, product label, and > MSD Sheet revision to do. > > Regards, > Mark Ray -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ > > John Kiernan wrote: > > >Point taken! I notice that Chroma do use > >the correct English spelling of fuchsine. > >Like nearly everyone else they spell > >phloxin with a terminal e, which is wrong > >because it's in the same group as eosin. > >Chroma also put a wrong terminal e on their > >English spelling of erythrosin; that one is > >unusual. > > --- John Kiernan > >------------------------------------ > >Gudrun Lang wrote: > > > > > >>Dear John, > >>I think the German spelling is Fuchsin (without e). Please look at > >>www.chroma.de at their catalogue. > >>Your vixen is German "F?chsin" (fuechsin). > >> > >>greetings from Austria > >>Gudrun > >> > >>----- Original Message ----- > >>From: "John Kiernan" > >>To: "Rena" ; > >>Sent: Thursday, September 30, 2004 5:29 AM > >>Subject: Re: [Histonet] Red chromogens [alkaline phosphatase labels] > >> > >>Dear Rena Fail, > >> > >>Red alkaline phosphatase product. > >> > >>Methods giving permanently mountable red products have been available > >>for many years. > >>Davis & Ornstein (1959) introduced hexazotized pararosaniline as a > >>trifunctional trapping agent for naphthols released by hydrolysis of > >>naphthol phosphates. Pararosaniline (CI 42500; CI Basic red 9) is one of > >>the four components of basic fuchsine and is commercially available in > >>fairly pure form. New fuchsine (CI 42520; CI Basic violet 2), which is > >>closely similar to pararosaniline and also available in pure form > >>(Horobin, 2002); It is currently preferred for making the hexazotized > >>reagent. > >> > >>Kits are sold, but the substrate solution is easily made in the lab. In > >>a simple procedure published at the IHCWorld.com (2004) web site (see > >>below for its URL), the working mixture is prepared from four stock > >>solutions, which are stored at 4 C. Solution D is warmed to room > >>temperature before using. The amount of levamisole (inhibitor of > >>endogenous tissue alkaline phosphatase) is clearly stated. > >> > >>The working solution is made immediately before using. Mix 10 drops each > >>of A and B, then add 10 drops of solution C followed by 10 Kits are > >>sold, but the substrate solution is easily made in the lab. In a simple > >>procedure published at the IHCWorld.com (2004) web site, the working > >>mixture is prepared from four stock solutions, which are stored at 4 C. > >>Solution D is warmed to room temperature before using. I've annotated > >>the instructions from IHCworld.com to simplify the weighing and > >>measuring. > >> > >>A. New fuchsine (CI 42520), 0.2% in 2M HCl. (Conc. HCl is 10 or 12M) > >>B. Sodium nitrite (NaNO2), 0.4% and levamisole,(= [-]-tetramisole > >>hydrochloride) 0.48% in water. > >>C. Naphthol AS-BI phosphate, 1% in 100% dimethylformamide. > >>D. 0.05 M Tris-HCl buffer, pH 8.7. > >> > >>The working solution is made immediately before using. Mix 10 drops each > >>of A and B, then add 10 drops of solution C followed by 10ml of the > >>buffer (D). A drop is assumed to be 0.05 ml. > >> > >>Sections are incubated in the working solution for 10-20 minutes at room > >>temperature. They are then washed counterstained as desired, dehydrated, > >>cleared and coverslipped. > >> > >>Lojda et al (1979, p. 74-75) warn that the working solution may be red > >>or brown from colored impurities in new fuchsine or pararosaniline. Such > >>solutions cause excessive yellow background staining. The yellow/brown > >>impurities are the same ones that can make Schiff's reagent yellow > >>(removable from Schiff with activated charcoal). Pararosaniline > >>certified by the Biological Stain Commission is suitable for making > >>alkaline phosphatase substrate mixtures because it is required to be > >>substantially free of brown and yellow materials (Penney et al., 2002). > >> > >>Probably any batch of basic fuchsine certified by the Biological Stain > >>Commission will be OK for making the IHCWorld.com alkaline phosphatase > >>substrate solution, because each of the four possible components of > >>basic fuchsine has three diazotizable amino groups and their molecular > >>weights are all quite close. I don't know of any published study that > >>establishes this, so you're probably safest to go with either certified > >>pararosaniline or with new fuchsine from a reputable dealer. > >> > >>References. (Please check them out if you can. The 2nd is easy enough!) > >> > >>Davis & Ornstein (1959) J. Histochem. Cytochem. 7:297-298. > >> > >>IHCWorld.com (2004) New fuchsin alkaline phosphatase substrate solution. > >>http://www.ihcworld.com/_protocols/chromogen_substrates/AP_new_fuchsin_red.htm > >> > >>Horobin (2002) Chapter 14 in Conn's Biological Stains, 10th ed. > >>Oxford:BIOS. > >> > >>Lojda et al (1979) Enzyme Histochemistry. Berlin: Springer. > >> > >>Penney et al. (2002) Biotech. Histochem. 77:237-275. > >> > >>Finally, please note that the correct spelling is fuchsine, not fuchsin, > >>despite anything you might read in a catalog or on a bottle label. This > >>is not a matter of American, British and German spellings. (Fuchsin is > >>German for vixen; the dyes are, I think, named from their colours being > >>similar to those of Fuchsia flowers rather than foxes.) Look in > >>Websters, the Oxford, etc. It's fuchsine with an e. An -ine ending of an > >>informal name of a chemical indicates that it is a base, often an amine. > >>The -in ending occurs with compounds that are not bases, such as > >>dextrin, eosin etc. > >> > >> John Kiernan > >> London, Canada > >>------------------------------------------------------------------ > >>Rena wrote: > >> > >> > >>>I have been having some trouble with some Abs stained with New Fuchsin > >>>and permanent red. Recently I was asked if I were using Levamisole in > >>>the permanent red and if so how much? I was told too much Levamisole in > >>>permanent red would result in no staining. I have used 1 drop per ml for > >>>both New Fuchsin and permanent red, which is the appropriate amount. We > >>>have run both DABS and either permanent red or New Fuchsin with the > >>>same AB from the same vial with vastly different results. Any ideas? > >>>Rena Fail > >>>_______________________________________________ > >>>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 > >> > >> > > > > > > Mark Ray wrote: > > John Kiernan wrote: > > >Point taken! I notice that Chroma do use > >the correct English spelling of fuchsine. > >Like nearly everyone else they spell > >phloxin with a terminal e, which is wrong > >because it's in the same group as eosin. > >Chroma also put a wrong terminal e on their > >English spelling of erythrosin; that one is > >unusual. > > --- John Kiernan > >------------------------------------ > >Gudrun Lang wrote: > > > > > >>Dear John, > >>I think the German spelling is Fuchsin (without e). Please look at > >>www.chroma.de at their catalogue. > >>Your vixen is German "F?chsin" (fuechsin). > >> > >>greetings from Austria > >>Gudrun > >> > >>----- Original Message ----- > >>From: "John Kiernan" > >>To: "Rena" ; > >>Sent: Thursday, September 30, 2004 5:29 AM > >>Subject: Re: [Histonet] Red chromogens [alkaline phosphatase labels] > >> > >>Dear Rena Fail, > >> > >>Red alkaline phosphatase product. > >> > >>Methods giving permanently mountable red products have been available > >>for many years. > >>Davis & Ornstein (1959) introduced hexazotized pararosaniline as a > >>trifunctional trapping agent for naphthols released by hydrolysis of > >>naphthol phosphates. Pararosaniline (CI 42500; CI Basic red 9) is one of > >>the four components of basic fuchsine and is commercially available in > >>fairly pure form. New fuchsine (CI 42520; CI Basic violet 2), which is > >>closely similar to pararosaniline and also available in pure form > >>(Horobin, 2002); It is currently preferred for making the hexazotized > >>reagent. > >> > >>Kits are sold, but the substrate solution is easily made in the lab. In > >>a simple procedure published at the IHCWorld.com (2004) web site (see > >>below for its URL), the working mixture is prepared from four stock > >>solutions, which are stored at 4 C. Solution D is warmed to room > >>temperature before using. The amount of levamisole (inhibitor of > >>endogenous tissue alkaline phosphatase) is clearly stated. > >> > >>The working solution is made immediately before using. Mix 10 drops each > >>of A and B, then add 10 drops of solution C followed by 10 Kits are > >>sold, but the substrate solution is easily made in the lab. In a simple > >>procedure published at the IHCWorld.com (2004) web site, the working > >>mixture is prepared from four stock solutions, which are stored at 4 C. > >>Solution D is warmed to room temperature before using. I've annotated > >>the instructions from IHCworld.com to simplify the weighing and > >>measuring. > >> > >>A. New fuchsine (CI 42520), 0.2% in 2M HCl. (Conc. HCl is 10 or 12M) > >>B. Sodium nitrite (NaNO2), 0.4% and levamisole,(= [-]-tetramisole > >>hydrochloride) 0.48% in water. > >>C. Naphthol AS-BI phosphate, 1% in 100% dimethylformamide. > >>D. 0.05 M Tris-HCl buffer, pH 8.7. > >> > >>The working solution is made immediately before using. Mix 10 drops each > >>of A and B, then add 10 drops of solution C followed by 10ml of the > >>buffer (D). A drop is assumed to be 0.05 ml. > >> > >>Sections are incubated in the working solution for 10-20 minutes at room > >>temperature. They are then washed counterstained as desired, dehydrated, > >>cleared and coverslipped. > >> > >>Lojda et al (1979, p. 74-75) warn that the working solution may be red > >>or brown from colored impurities in new fuchsine or pararosaniline. Such > >>solutions cause excessive yellow background staining. The yellow/brown > >>impurities are the same ones that can make Schiff's reagent yellow > >>(removable from Schiff with activated charcoal). Pararosaniline > >>certified by the Biological Stain Commission is suitable for making > >>alkaline phosphatase substrate mixtures because it is required to be > >>substantially free of brown and yellow materials (Penney et al., 2002). > >> > >>Probably any batch of basic fuchsine certified by the Biological Stain > >>Commission will be OK for making the IHCWorld.com alkaline phosphatase > >>substrate solution, because each of the four possible components of > >>basic fuchsine has three diazotizable amino groups and their molecular > >>weights are all quite close. I don't know of any published study that > >>establishes this, so you're probably safest to go with either certified > >>pararosaniline or with new fuchsine from a reputable dealer. > >> > >>References. (Please check them out if you can. The 2nd is easy enough!) > >> > >>Davis & Ornstein (1959) J. Histochem. Cytochem. 7:297-298. > >> > >>IHCWorld.com (2004) New fuchsin alkaline phosphatase substrate solution. > >>http://www.ihcworld.com/_protocols/chromogen_substrates/AP_new_fuchsin_red.htm > >> > >>Horobin (2002) Chapter 14 in Conn's Biological Stains, 10th ed. > >>Oxford:BIOS. > >> > >>Lojda et al (1979) Enzyme Histochemistry. Berlin: Springer. > >> > >>Penney et al. (2002) Biotech. Histochem. 77:237-275. > >> > >>Finally, please note that the correct spelling is fuchsine, not fuchsin, > >>despite anything you might read in a catalog or on a bottle label. This > >>is not a matter of American, British and German spellings. (Fuchsin is > >>German for vixen; the dyes are, I think, named from their colours being > >>similar to those of Fuchsia flowers rather than foxes.) Look in > >>Websters, the Oxford, etc. It's fuchsine with an e. An -ine ending of an > >>informal name of a chemical indicates that it is a base, often an amine. > >>The -in ending occurs with compounds that are not bases, such as > >>dextrin, eosin etc. > >> > >> John Kiernan > >> London, Canada > >>------------------------------------------------------------------ > >>Rena wrote: > >> > >> > >>>I have been having some trouble with some Abs stained with New Fuchsin > >>>and permanent red. Recently I was asked if I were using Levamisole in > >>>the permanent red and if so how much? I was told too much Levamisole in > >>>permanent red would result in no staining. I have used 1 drop per ml for > >>>both New Fuchsin and permanent red, which is the appropriate amount. We > >>>have run both DABS and either permanent red or New Fuchsin with the > >>>same AB from the same vial with vastly different results. Any ideas? > >>>Rena Fail > >>>_______________________________________________ > >>>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 > >> > >> > > > > > > From lu_ze <@t> sbcglobal.net Tue Oct 5 15:12:40 2004 From: lu_ze <@t> sbcglobal.net (Ze Lu) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Fisher 266mp tissue processor Err 03, any help Message-ID: <027401c4ab17$aa907d50$1302a8c0@optimum2> Hello, histonet friends, We had a quite old Fisher 266mp tissue processor in our lab. Every time when we start the program cycle, it showed Err 03 on screen. only very low level of solution go into the process pot. From the manual, it is indicated as overflow. We are quite sure that we did not load a lot of into tank. We also check the vacuum ortice in the process pot, and it is not blocked. The rotation valve is aldo fine when we checked. Could it be a problem of vacuum pump or leaking in some tubing? We appreciate your help. Ze Lu Optimum Therapeutics, LLC From juan.gutierrez <@t> christushealth.org Tue Oct 5 12:24:23 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] tissue processors Message-ID: Nothing beats the Tissue Teks. They keep going and going and going. Sorry but I can not say anything bad about equipment or companies anymore. Too many damn lawyers here in the USofA. Juan -----Original Message----- From: Andres Kulla [mailto:Andres.Kulla@kliinikum.ee] Sent: Tuesday, October 05, 2004 1:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] tissue processors To the experts in automated tissue processing, Could anybody share his/her experience with TBS (Triangle Biomedical Sciences) ATP1 tissue processor. Positive vs negative aspects in comparison with other devices eg Sakuras Tissue Tek VIP 500. With best regards from Andres Kulla, Estonia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jameel.syed <@t> abbott.com Tue Oct 5 12:29:01 2004 From: jameel.syed <@t> abbott.com (jameel.syed@abbott.com) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Re: mouse paws Message-ID: We process paws routinely. We haven't felt the need to deglove the skin. However, we decal them briefly and trim the foot pad longitudinally on both sides. After trimming we complete the decaling and process them with a program that is slightly longer paraffin infiltration. Trimming the skin off greatly expedites decaling and improves infiltration. In case of highly arthritic samples it makes them small enough to fit in a cassette for processing. Good luck, Jameel Syed, BS, HT/HTL, QIHC (ASCP) Pathology Supervisor Abbott BioResearch Center 100 Research Drive, Worcester, MA 01605 Tel: (508)849-2862 Fax: (508)793-4895 --------------------------------------------------------------------- Message: 1 Date: Tue, 05 Oct 2004 08:30:42 -0600 From: Gayle Callis Subject: Re: [Histonet] mouse paws To: "joyce judge" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20041005082945.01b0f408@gemini.msu.montana.edu> Content-Type: text/plain; charset="iso-8859-1"; format=flowed No we leave paws intact. However, mouse paws are extremely dense even though tiny, and after decalcification, we do a longer processing schedule. At 05:15 AM 10/5/2004, you wrote: >I would like too know if people that are processing mouse paws deglove or >process with the skin on. > >Joyce Judge >Visen Medical >jjudge@visenmedical.com > >_________________________________________________________________ >Is your PC infected? Get a FREE online computer virus scan from McAfee? >Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) *************************************** From Virginia.Achstetter <@t> afip.osd.mil Tue Oct 5 12:45:11 2004 From: Virginia.Achstetter <@t> afip.osd.mil (Achstetter, Virginia A.) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] (no subject) Message-ID: <9C631520464F9E4BA5B111450F2A0AAD0BCAF3@lewis.afip.osd.mil> Does anyone have a good protocol for cutting Microarray blocks? I understand that there are different methods of sealing the block cores so they stay put when sectioning. Ginny Achstetter HT (ASCP) Armed Forces Institute of Pathology Soft Tissue Pathology 6825 16th St. NW Bldg 54 Rm. 3062 Washington, DC 20306 Fax: 202-782-9182 Phone: 202-782-1914 From dpahisto <@t> yahoo.com Tue Oct 5 12:53:58 2004 From: dpahisto <@t> yahoo.com (Cindy DuBois) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Mounting media/ethanol Message-ID: <20041005175358.18251.qmail@web41303.mail.yahoo.com> We use Clearium from Surgipath and coverslip out of Isopropyl alcohol. We have been doing this for several years. One drawback we have seen is if they dried to quickly (ie. put in oven for drying), we see lots of bubbles. We just put the flats on top of the embedding center for about 15 minuts and they are dry enough for the docs. Then when we get them back we let them sit at room temp overnight, then put in over (60 C) overnight. Another drawback is when using it with any alcohol soluble stain (Mucicarmine, Gram, etc.) the stain will leach out over time. On these slides, we merely coverslip out of xylene. Hope this helps, Cindy DuBois, HT ASCP Delta Pathology Stockton CA --------------------------------- Do you Yahoo!? Yahoo! Mail Address AutoComplete - You start. We finish. From Julie.Sanders <@t> med.va.gov Tue Oct 5 12:53:26 2004 From: Julie.Sanders <@t> med.va.gov (Sanders, Julie, VHACIN) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] HPV Ventana Benchmark Message-ID: <457381D92B01BD44B21CF37CC02EBDFD28E97A@vhacinexc2.v10.med.va.gov> Are you looking for a quote for the Benchmark itself, or the cost of doing HPV's? You can contact Ventana to have a sales rep visit your site if looking for the machine. If you want cost for doing HPV's, I could figure up what it is costing us. Julie Julie Sanders, BA, HT(ASCP) Supervisor, Anatomic Pathology VAMC, Cincinnati, Oh. From lizchlipala <@t> premierhistology.com Tue Oct 5 13:07:57 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] mouse paws In-Reply-To: Message-ID: <002601c4ab06$3e5ef320$76d48a80@AMY> We routinely process mouse paws for arthritis studies and we do not take the skin off. This way the specimens remain intact. I agree with Gayle and that we also process on a longer processing cycle also. There are pictures on our web site of mouse paws and ankles from arthritis studies, just go to the gallery. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Histology Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 lizchlipala@premierhistology.com www.premierhistology.com Ship to Address: Premier Histology Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of joyce judge Sent: Tuesday, October 05, 2004 4:16 AM To: histonet@pathology.swmed.edu Subject: [Histonet] mouse paws I would like too know if people that are processing mouse paws deglove or process with the skin on. Joyce Judge Visen Medical jjudge@visenmedical.com _________________________________________________________________ Is your PC infected? Get a FREE online computer virus scan from McAfeeR Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eva.alstromer <@t> histolab.se Tue Oct 5 13:32:12 2004 From: eva.alstromer <@t> histolab.se (=?iso-8859-1?Q?Eva_Alstr=F6mer?=) Date: Fri Sep 16 15:24:07 2005 Subject: SV: [Histonet] Mounting media to covberslip from ethano In-Reply-To: <4162C90E.6020303@shaw.ca> Message-ID: <001501c4ab09$a3521f20$ba9843d5@HISSWE.lokal> Dear Paul, In Europ? it is two different mounting mediums(containing xylene), Mountex and Pertex in the market. Both give you possibility to mount direct after an extra jar with clean alcohol, these two mountingmedium are very fast drying and do not change in colour after storing of many years. Eva -----Ursprungligt meddelande----- Fr?n: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] F?r Paul Bradbury Skickat: den 5 oktober 2004 18:17 Till: HistoNet Server ?mne: Re: [Histonet] Mounting media to covberslip from ethano I have never encountered a mounting medium that uses alcohol as the solvent. If one exists, two disadvantages spring to mind immediately. One, the refractive index of the medium would be low, so the clarity and brilliance associated with conventional, xylene-based media would be absent. Two, a large proportion of counterstains (eosin, van Gieson, neutral red, methylene blue, etc) are alcohol -soluble and would "bleed" into the medium. Paul _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From n_hedgecock <@t> hotmail.com Tue Oct 5 13:42:45 2004 From: n_hedgecock <@t> hotmail.com (Nicole Hedgecock) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] decalcified sections of bone Message-ID: Hi, All I am attempting to do TUNEL staining of osteocytes in cortical bone. I was wondering why the bone has to be decalcified in order to do this process. And Why do the sections have to be so thin? Thanks Nicole Nicole Hedgecock Master's Student Orthopedic Research Lab UCD Medical Center University of California, Davis _________________________________________________________________ ?Cu?nto vale tu auto? Tips para mantener tu carro. ?De todo en MSN Latino Autos! http://latino.msn.com/autos/ From gcallis <@t> montana.edu Tue Oct 5 15:11:44 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Attn: Ann Featherstone, failed message delivery Message-ID: <6.0.0.22.1.20041005141005.01b0a4b8@gemini.msu.montana.edu> To Ann Featherstone: Message with mast cell staining protocols could not be delivered to your address, recontact me to get them again. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From vazquezr <@t> ohsu.edu Tue Oct 5 15:59:13 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] tissue processing Message-ID: Histoneters, I do microwave tissue processing, when tissue comes out, after the paraffin and it is still mushy and has the feel of a sponge. Does it mean that I should keep in isopropyl alittle longer to make it alittle firmer and that it isn't dehydrated all the way? And paraffin is having I hard time infiltrating the tissue? Robyn OHSU Portland, Or From tissuearray <@t> hotmail.com Tue Oct 5 16:01:10 2004 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Tissuearrays Message-ID: Hi Virgina, Have you looked on my website. I have several articles in the journal of histotechnology and some instructional videos on the same subjects that go into detail on the subject of array construction. My website is www.arrayworkshop.com 1. What I do is place the array block on a slide face down. Be sure to put the slide on it before you flip the block over. Just in case some of the cores fall out of the block depending on how loose the cores are. 2. Put the block ans slide (slide down) in an oven at approx. 37-40 'C for 15 to 20 minutes. 3. Press the slide and block together. You will notice that the paraffin will melt a little and the paraffin will spread out slightly. This is good, because this is how the punches set into the block. 4. DO NOT seperate the slide and block at this time. Place then together on an ice tray and allow to cool. The slide will seperate easily once they are cold. 5. Before cutting, trim the sides a little to make them straight and flat. This will help the ribboning to go smoother with flat edges. 6. Do not soak the block in ice water. I have found that ice water makes the punches swell thus making the tissue mushey. Use only Ice. If you feel you might get a better cut with water only soak for a minute or so. 7. Use a fresh blade to make a ribbon. I even knock down the blade with a Kim wipe to help the cutting go quickly. Running the kim wipe across a new blade removes oils also. You will find the knife make ribbons right away. The rest is normal histology techniques. Lay the ribbons on the water bath, etc... If you have any more questions please feel free to email me. Thom Jensen HT (ASCP) / Array Technician >From: "Achstetter, Virginia A." >To: >Subject: [Histonet] (no subject) >Date: Tue, 5 Oct 2004 13:45:11 -0400 > >Does anyone have a good protocol for cutting Microarray blocks? I understand that there are different methods of sealing the block cores so they stay put when sectioning. > >Ginny Achstetter HT (ASCP) >Armed Forces Institute of Pathology >Soft Tissue Pathology >6825 16th St. NW >Bldg 54 Rm. 3062 >Washington, DC 20306 >Fax: 202-782-9182 >Phone: 202-782-1914 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ [1]Rock, jazz, country, soul & more. Find the music you love on MSN Music! References 1. http://g.msn.com/8HMAENUS/2740??PS=47575 From spoulos <@t> saa.ars.usda.gov Tue Oct 5 16:42:55 2004 From: spoulos <@t> saa.ars.usda.gov (Sylvia Poulos) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] NADPH-TR protocol Message-ID: Hey all, Does anyone have a protocol for NADPH-tetrazolium reductase staining on frozen tissues that they'd be willing to send me? As always, thanks for all of the help! Sylvia Sylvia P. Poulos USDA-ARS-Animal Physiology Research Unit Athens, GA 30605 706-583-8279 706-542-0399 (fax) From n_hedgecock <@t> hotmail.com Tue Oct 5 16:44:50 2004 From: n_hedgecock <@t> hotmail.com (Nicole Hedgecock) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] More details: decalcified sections of bone Message-ID: I sent this out earlier (see below) and more details of my tissue processing and sectioning were requested. I want to identify apoptotic osteocytes in rabbit cortical bone as well as some morphological features especially microcracks. I fix the bones in 4% paraformaldehyde for 24 hours, then place then in 70% EtOH until I can decalcify them in EDTA at room temp. Between the EtOH and EDTA steps I rinse the bones in dH2O for about 3 days. After decalcification, the bones are again rinsed in dH2O and then embedded in paraffin and sectioned to 6 microns. I use the TUNEL kit from Roche for In situ labeling of DNA breaks. Do you think that I have to do decalcification for the TUNEL to work? Does sectioning of paraffin embedded tissue create a lot of microscopic damage to the section? I hope this clears up my situation and goals. Nicole >From: "Nicole Hedgecock" >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] decalcified sections of bone >Date: Tue, 05 Oct 2004 14:42:45 -0400 > >Hi, All > >I am attempting to do TUNEL staining of osteocytes in cortical bone. I was >wondering why the bone has to be decalcified in order to do this process. >And Why do the sections have to be so thin? > >Thanks >Nicole > > > >Nicole Hedgecock >Master's Student >Orthopedic Research Lab >UCD Medical Center >University of California, Davis > >_________________________________________________________________ >?Cu?nto vale tu auto? Tips para mantener tu carro. ?De todo en MSN Latino >Autos! http://latino.msn.com/autos/ > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Visita MSN Latino Entretenimiento: ?m?sica, cine, chismes, TV y m?s...! http://latino.msn.com/entretenimiento/ From lkbauer <@t> unmc.edu Tue Oct 5 16:53:39 2004 From: lkbauer <@t> unmc.edu (lkbauer@unmc.edu) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] orienting mouse embryos Message-ID: Could I tap your collective experience to help solve a problem? I am trying to orient gestational day 9.5 mouse embryos in paraffin for cross sectioning. After processing, these specimens are like pieces of fragil white pepper. I can only see head from tail with a microscope. By the time I get everything in focus the parffin is turning white....like finding a ghost in a blizzard. Does anyone have any tips for this task? Thanks, Lin Linda(Lin)Bauer Department of Genetics, Cell Biology, and Anatomy University of Nebraska Medical Center Omaha, NE 68198-5455 Email: lkbauer@unmc.edu From Janet.Bonner <@t> FLHOSP.ORG Tue Oct 5 17:10:01 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] orienting mouse embryos Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4092@fh2k093.fhmis.net> Put eosin-phloxine stain (80ml to 3500ml) in your first alcohol when processing, we use this on our biopsies and it makes all the difference!. Janet -----Original Message----- From: lkbauer@unmc.edu [mailto:lkbauer@unmc.edu] Sent: Tuesday, October 05, 2004 5:54 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] orienting mouse embryos Could I tap your collective experience to help solve a problem? I am trying to orient gestational day 9.5 mouse embryos in paraffin for cross sectioning. After processing, these specimens are like pieces of fragil white pepper. I can only see head from tail with a microscope. By the time I get everything in focus the parffin is turning white....like finding a ghost in a blizzard. Does anyone have any tips for this task? Thanks, Lin Linda(Lin)Bauer Department of Genetics, Cell Biology, and Anatomy University of Nebraska Medical Center Omaha, NE 68198-5455 Email: lkbauer@unmc.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From laurie.reilly <@t> jcu.edu.au Tue Oct 5 17:18:08 2004 From: laurie.reilly <@t> jcu.edu.au (Laurie Reilly) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] orienting mouse embryos In-Reply-To: Message-ID: <5.1.0.14.0.20041006081451.00a39b30@mail.jcu.edu.au> Dear Lin, Try putting a few mls of aqueous eosin in your fixative. A pink specimen is a lot easier to see. The eosin can be washed out at a later stage if it is not required. Good luck and regards, Laurie. At 04:53 PM 10/05/04 -0500, lkbauer@unmc.edu wrote: >Could I tap your collective experience to help solve a problem? I am trying >to orient gestational day 9.5 mouse embryos in paraffin for cross >sectioning. After processing, these specimens are like pieces of fragil >white pepper. I can only see head from tail with a microscope. By the time >I get everything in focus the parffin is turning white....like finding a >ghost in a blizzard. Does anyone have any tips for this task? Thanks, Lin > >Linda(Lin)Bauer >Department of Genetics, Cell Biology, and Anatomy >University of Nebraska Medical Center >Omaha, NE 68198-5455 >Email: lkbauer@unmc.edu > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mr.Laurie Reilly Ph 07 4781 4468 Physiology & Pharmacology Fax 07 4779 1526 Aust.Inst.of Tropical Vet.& Animal Sc. James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. From ajennings <@t> unmc.edu Tue Oct 5 17:15:33 2004 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] orienting mouse embryos Message-ID: Lin Run downstairs and I will show you the tricks of the trade.....thought I had already shown you :) From Gervaip <@t> aol.com Tue Oct 5 18:01:18 2004 From: Gervaip <@t> aol.com (Gervaip@aol.com) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Reusing the filter for ThinPrep for HPV Message-ID: <110.398ec1e3.2e9481be@aol.com> Is anyone in HistoLand doing this? We were told we can reuse the filter to prepare another slide for HPV. If you do, how do you dry the filter? Pearl in New Orleans From darrenj <@t> medica.co.nz Tue Oct 5 18:51:05 2004 From: darrenj <@t> medica.co.nz (Darren James) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] Bone Saw and Dust Extraction Message-ID: <000e01c4ab36$2e3140e0$c864a8c0@medica.co.nz> Hello, I have a question, does anyone know if there is an electric bone saw available complete with a dust extraction unit for a Histology lab? If not, would a saw with a water spray eliminate aerosol and particulate matter? I am looking for a benchtop saw for a histo lab not a Stryker or similar mortuary saw. Thanks Darren James Technical Representative Medica Pacifica Ltd Office Address: 1A 153 Stoddard Road, Mt Roskill, Auckland Postal Address: PO Box 24-421, Royal Oak, Auckland 1030 Ph: +64 9 629 0823 Fax: +64 9 629 0542 Mob: + 64 21 633 820 From rockbeki <@t> ufl.edu Tue Oct 5 22:01:49 2004 From: rockbeki <@t> ufl.edu (SMITH,REBEKAH FELICIA) Date: Fri Sep 16 15:24:07 2005 Subject: [Histonet] DAB-colors... Message-ID: <119304449.1097031709998.JavaMail.osg@osgjas02.cns.ufl.edu> The only two colors I know of for DAB are brown and black. The black is due to adding nickel to DAB, I think. I think pierce sells them. On Tue Oct 05 10:48:28 EDT 2004, "Louro, Pedro" wrote: > Hello all, > > When at NSH -Toronto..I heard people talking about using DAB in > different > colors. > Can someone tell who sells this product. > > Thanks Pedro > > > ********************************************************************* > This message and any attachments are solely for the intended > recipient. If you are not the intended recipient, disclosure, > copying, use or distribution of the information included in this > message is prohibited -- Please immediately and permanently > delete. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" From jluis.palazon <@t> icman.csic.es Wed Oct 6 01:46:37 2004 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] elastic fibers Message-ID: <20041006064637.2BF5831C921@perceval.uca.es> Dear List fellows Many thanks for all of you who answered to my question on elastic fibers stain. As I can see the more popular stains are in order of preference 1)Verhoeff's Van Gieson 2) Weigher resorcin fuchsin. Greetings Jos? Luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es From BMolinari <@t> heart.thi.tmc.edu Wed Oct 6 06:16:45 2004 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] processing question Message-ID: Hi histonetters, Is it harmful to tissue that is in 70% ETOH to be put on the processor and start at 10% NBF? Thanks . Betsy Molinari HT(ASCP) Texas Heart Institute Houston,TX 77030 832-355-6524 From JNocito <@t> Pathreflab.com Wed Oct 6 08:18:46 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Reusing the filter for ThinPrep for HPV In-Reply-To: <110.398ec1e3.2e9481be@aol.com> Message-ID: Pearl, we routinely put the patient's filter in the Preservecyte vial after the Thin prep is made. If we need o go back to that vial, we empty whatever material is in the filter, back into the vial. Very carefully, we wipe both sides of the filter with a Kimwipe and let the filter air dry (about a minute. We try not to put too much pressure on the filter because sometimes it comes off. Depending on the cellularity of the specimen, I have been able to get 3 additional slides before I had to get another filter. I think they clog up after a while. Now, if anyone from Cytyc is listening, my rep is Billy Redding. Disclaimer: This method is not approved by Cytyc, the FDA, the FCC, the FBI and the CIA, the GOP, nor the DNC. I will be expecting a phone call from Billy any minute now. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gervaip@aol.com Sent: Tuesday, October 05, 2004 6:01 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Reusing the filter for ThinPrep for HPV Is anyone in HistoLand doing this? We were told we can reuse the filter to prepare another slide for HPV. If you do, how do you dry the filter? Pearl in New Orleans _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From plucas <@t> biopath.org Wed Oct 6 08:49:19 2004 From: plucas <@t> biopath.org (Paula Lucas) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Cassette Label System Message-ID: <01C4AB70.9B2AB250.plucas@biopath.org> Hello, My company is possibly interested in purchasing a histology cassette printer. Could you please provide information on the different manufactures that offer printers? What are the pro/cons? Is it really that worth it to purchase one? Does it really save that much time? Are they easy to use? Any information, suggestions, and comments are greatly appreciated. Thank you, Paula Lucas Bio-Path Medical Group From jbrod <@t> tvmdl.tamu.edu Wed Oct 6 08:48:23 2004 From: jbrod <@t> tvmdl.tamu.edu (Jordan Brod) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Eye Protection Question Message-ID: Good Morning! What kind of eye protection are technicians required to wear in the laboratory during grossing, sectioning, processor maintenance, etc.? Thanks and have a great day. Jordan TVMDL - College Station, TX From Jackie.O'Connor <@t> abbott.com Wed Oct 6 09:22:40 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Eye Protection Question Message-ID: Protection from biohaz splash where appropriate, protection from chemical splash, where appropriate. As a note - just to walk into our labs here, even just to say hi to another person, we must have our safety glasses and labcoats on. The thinking is that someone else working may create a splash that sails across the room into your eye. "Jordan Brod" Sent by: histonet-bounces@lists.utsouthwestern.edu 10/06/2004 08:48 AM To: cc: Subject: [Histonet] Eye Protection Question Good Morning! What kind of eye protection are technicians required to wear in the laboratory during grossing, sectioning, processor maintenance, etc.? Thanks and have a great day. Jordan TVMDL - College Station, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Wed Oct 6 10:55:37 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Cassette Label System In-Reply-To: <01C4AB70.9B2AB250.plucas@biopath.org> Message-ID: Paula, we just purchased a cassette printer from Surgipath. It prints one block at a time, but can be set to automatically increment. We tested one printer from TBS, but the cassettes kept jamming. The Surgipath printer maybe slower, but it has less moving parts to break down and it does a great job. Disclaimer: As usual, the opinions of the author in no way reflect the opinions of the company, it executives or relatives. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Paula Lucas Sent: Wednesday, October 06, 2004 8:49 AM To: Histonet (E-mail) Subject: [Histonet] Cassette Label System Hello, My company is possibly interested in purchasing a histology cassette printer. Could you please provide information on the different manufactures that offer printers? What are the pro/cons? Is it really that worth it to purchase one? Does it really save that much time? Are they easy to use? Any information, suggestions, and comments are greatly appreciated. Thank you, Paula Lucas Bio-Path Medical Group _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Wed Oct 6 10:59:39 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Eye Protection Question In-Reply-To: Message-ID: Jordon, we have full-face safety shields as well as surgical masks with an attached eye shield. We wear either when changing the staining machines and tissue processors. When I gross, I wear my eyeglasses that are classified as safety wear. We have never had an incident, but we flush our eyewash stations weekly because you never know what will happen. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jordan Brod Sent: Wednesday, October 06, 2004 8:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Eye Protection Question Good Morning! What kind of eye protection are technicians required to wear in the laboratory during grossing, sectioning, processor maintenance, etc.? Thanks and have a great day. Jordan TVMDL - College Station, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From fjones <@t> namsa.com Wed Oct 6 11:40:36 2004 From: fjones <@t> namsa.com (Fawn Jones) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] micronucleus staining Message-ID: <915E55B02E236E4D95258B181EEF6317075D2B@namsams01.namsa.int> Hi everyone, Our histology labs is doing a wright-giemsa stain on mice micronucleus slides and we are having a problem with our cells not picking up the stain. Are there any suggestions for us. We let them dry overnight then we put them in Methanol for 5 minutes, let them air dry for 20 minutes then they go into Wright's stain for 6 minutes, then Wright-giemsa stain for 6 minutes, and then go through 3 changes of sterile water for injection for 30 seconds each. Then they sit overnight before coverslipping. If anyone could help that would be great. Thanks Fawn Jones North American Science Associates From fjones <@t> namsa.com Wed Oct 6 12:07:41 2004 From: fjones <@t> namsa.com (Fawn Jones) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Micronucleus staining Message-ID: <915E55B02E236E4D95258B181EEF6317075D2C@namsams01.namsa.int> We need help with our Wright-Giemsa stain we are performing on our micronucleus slides. The cells are not picking up the stain and we are not sure why. Does any one have any suggestions or can send us their protocol they follow? E-mail me at fjones@namsa.com. Thanks Fawn Jones From KMH.02 <@t> ex.uchs.org Wed Oct 6 12:16:44 2004 From: KMH.02 <@t> ex.uchs.org (Hopkins, Karen) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] RE: Histonet Digest, Vol 11, Issue 7 Message-ID: <640B23A5DC4B234BB065E56F2DB3059618B8FC@uchex2ucmc.uchs.org> Comment about thinprep filter....I was also curious if anyone has done any "validation" studies to compare using the blue non-gyn filter instead of the clear gyn filter for gyn specimens. Apparantly the filter pores are smaller on the blue one so it should be okay- the problem is that the thinprep test for gyn's is an FDA approved test for use as the manufacturers instructions and to use the blue one would mean having to validate the results. I was curious because the cost of the blue filters is much less than the clear ones. Any thoughts? -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Wednesday, October 06, 2004 1:05 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 11, Issue 7 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Fisher 266mp tissue processor Err 03, any help (Ze Lu) 2. RE: tissue processors (GUTIERREZ, JUAN) 3. Re: mouse paws (jameel.syed@abbott.com) 4. (no subject) (Achstetter, Virginia A.) 5. Mounting media/ethanol (Cindy DuBois) 6. HPV Ventana Benchmark (Sanders, Julie, VHACIN) 7. RE: mouse paws (Elizabeth Chlipala) 8. SV: [Histonet] Mounting media to covberslip from ethano (Eva Alstr?mer) 9. decalcified sections of bone (Nicole Hedgecock) 10. Attn: Ann Featherstone, failed message delivery (Gayle Callis) 11. tissue processing (Robyn Vazquez) 12. RE: Tissuearrays (Thom Jensen) 13. NADPH-TR protocol (Sylvia Poulos) 14. More details: decalcified sections of bone (Nicole Hedgecock) 15. orienting mouse embryos (lkbauer@unmc.edu) 16. RE: orienting mouse embryos (Bonner, Janet) 17. Re: orienting mouse embryos (Laurie Reilly) 18. Re: orienting mouse embryos (ajennings@unmc.edu) 19. Reusing the filter for ThinPrep for HPV (Gervaip@aol.com) 20. Bone Saw and Dust Extraction (Darren James) 21. Re: DAB-colors... (SMITH,REBEKAH FELICIA) 22. elastic fibers (Jose Luis Palazon Fernandez) 23. processing question (Molinari, Betsy) 24. RE: Reusing the filter for ThinPrep for HPV (Joe Nocito) 25. Cassette Label System (Paula Lucas) 26. Eye Protection Question (Jordan Brod) 27. Re: Eye Protection Question (Jackie.O'Connor@abbott.com) 28. RE: Cassette Label System (Joe Nocito) 29. RE: Eye Protection Question (Joe Nocito) 30. micronucleus staining (Fawn Jones) ---------------------------------------------------------------------- Message: 1 Date: Tue, 5 Oct 2004 13:12:40 -0700 From: "Ze Lu" Subject: [Histonet] Fisher 266mp tissue processor Err 03, any help To: Message-ID: <027401c4ab17$aa907d50$1302a8c0@optimum2> Content-Type: text/plain; charset="iso-8859-1" Hello, histonet friends, We had a quite old Fisher 266mp tissue processor in our lab. Every time when we start the program cycle, it showed Err 03 on screen. only very low level of solution go into the process pot. From the manual, it is indicated as overflow. We are quite sure that we did not load a lot of into tank. We also check the vacuum ortice in the process pot, and it is not blocked. The rotation valve is aldo fine when we checked. Could it be a problem of vacuum pump or leaking in some tubing? We appreciate your help. Ze Lu Optimum Therapeutics, LLC ------------------------------ Message: 2 Date: Tue, 5 Oct 2004 12:24:23 -0500 From: "GUTIERREZ, JUAN" Subject: RE: [Histonet] tissue processors To: "Andres Kulla" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Nothing beats the Tissue Teks. They keep going and going and going. Sorry but I can not say anything bad about equipment or companies anymore. Too many damn lawyers here in the USofA. Juan -----Original Message----- From: Andres Kulla [mailto:Andres.Kulla@kliinikum.ee] Sent: Tuesday, October 05, 2004 1:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] tissue processors To the experts in automated tissue processing, Could anybody share his/her experience with TBS (Triangle Biomedical Sciences) ATP1 tissue processor. Positive vs negative aspects in comparison with other devices eg Sakuras Tissue Tek VIP 500. With best regards from Andres Kulla, Estonia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Tue, 5 Oct 2004 13:29:01 -0400 From: jameel.syed@abbott.com Subject: [Histonet] Re: mouse paws To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" We process paws routinely. We haven't felt the need to deglove the skin. However, we decal them briefly and trim the foot pad longitudinally on both sides. After trimming we complete the decaling and process them with a program that is slightly longer paraffin infiltration. Trimming the skin off greatly expedites decaling and improves infiltration. In case of highly arthritic samples it makes them small enough to fit in a cassette for processing. Good luck, Jameel Syed, BS, HT/HTL, QIHC (ASCP) Pathology Supervisor Abbott BioResearch Center 100 Research Drive, Worcester, MA 01605 Tel: (508)849-2862 Fax: (508)793-4895 --------------------------------------------------------------------- Message: 1 Date: Tue, 05 Oct 2004 08:30:42 -0600 From: Gayle Callis Subject: Re: [Histonet] mouse paws To: "joyce judge" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20041005082945.01b0f408@gemini.msu.montana.edu> Content-Type: text/plain; charset="iso-8859-1"; format=flowed No we leave paws intact. However, mouse paws are extremely dense even though tiny, and after decalcification, we do a longer processing schedule. At 05:15 AM 10/5/2004, you wrote: >I would like too know if people that are processing mouse paws deglove or >process with the skin on. > >Joyce Judge >Visen Medical >jjudge@visenmedical.com > >_________________________________________________________________ >Is your PC infected? Get a FREE online computer virus scan from McAfee? >Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) *************************************** ------------------------------ Message: 4 Date: Tue, 5 Oct 2004 13:45:11 -0400 From: "Achstetter, Virginia A." Subject: [Histonet] (no subject) To: Message-ID: <9C631520464F9E4BA5B111450F2A0AAD0BCAF3@lewis.afip.osd.mil> Content-Type: text/plain; charset="iso-8859-1" Does anyone have a good protocol for cutting Microarray blocks? I understand that there are different methods of sealing the block cores so they stay put when sectioning. Ginny Achstetter HT (ASCP) Armed Forces Institute of Pathology Soft Tissue Pathology 6825 16th St. NW Bldg 54 Rm. 3062 Washington, DC 20306 Fax: 202-782-9182 Phone: 202-782-1914 ------------------------------ Message: 5 Date: Tue, 5 Oct 2004 10:53:58 -0700 (PDT) From: Cindy DuBois Subject: [Histonet] Mounting media/ethanol To: histonet@lists.utsouthwestern.edu Message-ID: <20041005175358.18251.qmail@web41303.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii We use Clearium from Surgipath and coverslip out of Isopropyl alcohol. We have been doing this for several years. One drawback we have seen is if they dried to quickly (ie. put in oven for drying), we see lots of bubbles. We just put the flats on top of the embedding center for about 15 minuts and they are dry enough for the docs. Then when we get them back we let them sit at room temp overnight, then put in over (60 C) overnight. Another drawback is when using it with any alcohol soluble stain (Mucicarmine, Gram, etc.) the stain will leach out over time. On these slides, we merely coverslip out of xylene. Hope this helps, Cindy DuBois, HT ASCP Delta Pathology Stockton CA --------------------------------- Do you Yahoo!? Yahoo! Mail Address AutoComplete - You start. We finish. ------------------------------ Message: 6 Date: Tue, 5 Oct 2004 12:53:26 -0500 From: "Sanders, Julie, VHACIN" Subject: [Histonet] HPV Ventana Benchmark To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <457381D92B01BD44B21CF37CC02EBDFD28E97A@vhacinexc2.v10.med.va.gov> Content-Type: text/plain; charset="iso-8859-1" Are you looking for a quote for the Benchmark itself, or the cost of doing HPV's? You can contact Ventana to have a sales rep visit your site if looking for the machine. If you want cost for doing HPV's, I could figure up what it is costing us. Julie Julie Sanders, BA, HT(ASCP) Supervisor, Anatomic Pathology VAMC, Cincinnati, Oh. ------------------------------ Message: 7 Date: Tue, 5 Oct 2004 12:07:57 -0600 From: "Elizabeth Chlipala" Subject: RE: [Histonet] mouse paws To: "'joyce judge'" , Message-ID: <002601c4ab06$3e5ef320$76d48a80@AMY> Content-Type: text/plain; charset="US-ASCII" We routinely process mouse paws for arthritis studies and we do not take the skin off. This way the specimens remain intact. I agree with Gayle and that we also process on a longer processing cycle also. There are pictures on our web site of mouse paws and ankles from arthritis studies, just go to the gallery. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Histology Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 lizchlipala@premierhistology.com www.premierhistology.com Ship to Address: Premier Histology Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of joyce judge Sent: Tuesday, October 05, 2004 4:16 AM To: histonet@pathology.swmed.edu Subject: [Histonet] mouse paws I would like too know if people that are processing mouse paws deglove or process with the skin on. Joyce Judge Visen Medical jjudge@visenmedical.com _________________________________________________________________ Is your PC infected? Get a FREE online computer virus scan from McAfeeR Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Tue, 5 Oct 2004 20:32:12 +0200 From: Eva Alstr?mer Subject: SV: [Histonet] Mounting media to covberslip from ethano To: "'Paul Bradbury'" Cc: histonet@lists.utsouthwestern.edu Message-ID: <001501c4ab09$a3521f20$ba9843d5@HISSWE.lokal> Content-Type: text/plain; charset="iso-8859-1" Dear Paul, In Europ? it is two different mounting mediums(containing xylene), Mountex and Pertex in the market. Both give you possibility to mount direct after an extra jar with clean alcohol, these two mountingmedium are very fast drying and do not change in colour after storing of many years. Eva -----Ursprungligt meddelande----- Fr?n: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] F?r Paul Bradbury Skickat: den 5 oktober 2004 18:17 Till: HistoNet Server ?mne: Re: [Histonet] Mounting media to covberslip from ethano I have never encountered a mounting medium that uses alcohol as the solvent. If one exists, two disadvantages spring to mind immediately. One, the refractive index of the medium would be low, so the clarity and brilliance associated with conventional, xylene-based media would be absent. Two, a large proportion of counterstains (eosin, van Gieson, neutral red, methylene blue, etc) are alcohol -soluble and would "bleed" into the medium. Paul _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Tue, 05 Oct 2004 14:42:45 -0400 From: "Nicole Hedgecock" Subject: [Histonet] decalcified sections of bone To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=iso-8859-1; format=flowed Hi, All I am attempting to do TUNEL staining of osteocytes in cortical bone. I was wondering why the bone has to be decalcified in order to do this process. And Why do the sections have to be so thin? Thanks Nicole Nicole Hedgecock Master's Student Orthopedic Research Lab UCD Medical Center University of California, Davis _________________________________________________________________ ?Cu?nto vale tu auto? Tips para mantener tu carro. ?De todo en MSN Latino Autos! http://latino.msn.com/autos/ ------------------------------ Message: 10 Date: Tue, 05 Oct 2004 14:11:44 -0600 From: Gayle Callis Subject: [Histonet] Attn: Ann Featherstone, failed message delivery To: Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20041005141005.01b0a4b8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed To Ann Featherstone: Message with mast cell staining protocols could not be delivered to your address, recontact me to get them again. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 11 Date: Tue, 05 Oct 2004 13:59:13 -0700 From: "Robyn Vazquez" Subject: [Histonet] tissue processing To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Histoneters, I do microwave tissue processing, when tissue comes out, after the paraffin and it is still mushy and has the feel of a sponge. Does it mean that I should keep in isopropyl alittle longer to make it alittle firmer and that it isn't dehydrated all the way? And paraffin is having I hard time infiltrating the tissue? Robyn OHSU Portland, Or ------------------------------ Message: 12 Date: Tue, 05 Oct 2004 21:01:10 +0000 From: "Thom Jensen" Subject: RE: [Histonet] Tissuearrays To: Virginia.Achstetter@afip.osd.mil, histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain Hi Virgina, Have you looked on my website. I have several articles in the journal of histotechnology and some instructional videos on the same subjects that go into detail on the subject of array construction. My website is www.arrayworkshop.com 1. What I do is place the array block on a slide face down. Be sure to put the slide on it before you flip the block over. Just in case some of the cores fall out of the block depending on how loose the cores are. 2. Put the block ans slide (slide down) in an oven at approx. 37-40 'C for 15 to 20 minutes. 3. Press the slide and block together. You will notice that the paraffin will melt a little and the paraffin will spread out slightly. This is good, because this is how the punches set into the block. 4. DO NOT seperate the slide and block at this time. Place then together on an ice tray and allow to cool. The slide will seperate easily once they are cold. 5. Before cutting, trim the sides a little to make them straight and flat. This will help the ribboning to go smoother with flat edges. 6. Do not soak the block in ice water. I have found that ice water makes the punches swell thus making the tissue mushey. Use only Ice. If you feel you might get a better cut with water only soak for a minute or so. 7. Use a fresh blade to make a ribbon. I even knock down the blade with a Kim wipe to help the cutting go quickly. Running the kim wipe across a new blade removes oils also. You will find the knife make ribbons right away. The rest is normal histology techniques. Lay the ribbons on the water bath, etc... If you have any more questions please feel free to email me. Thom Jensen HT (ASCP) / Array Technician >From: "Achstetter, Virginia A." >To: >Subject: [Histonet] (no subject) >Date: Tue, 5 Oct 2004 13:45:11 -0400 > >Does anyone have a good protocol for cutting Microarray blocks? I understand that there are different methods of sealing the block cores so they stay put when sectioning. > >Ginny Achstetter HT (ASCP) >Armed Forces Institute of Pathology >Soft Tissue Pathology >6825 16th St. NW >Bldg 54 Rm. 3062 >Washington, DC 20306 >Fax: 202-782-9182 >Phone: 202-782-1914 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ [1]Rock, jazz, country, soul & more. Find the music you love on MSN Music! References 1. http://g.msn.com/8HMAENUS/2740??PS=47575 ------------------------------ Message: 13 Date: Tue, 05 Oct 2004 17:42:55 -0400 From: "Sylvia Poulos" Subject: [Histonet] NADPH-TR protocol To: Message-ID: Content-Type: text/plain; charset=US-ASCII Hey all, Does anyone have a protocol for NADPH-tetrazolium reductase staining on frozen tissues that they'd be willing to send me? As always, thanks for all of the help! Sylvia Sylvia P. Poulos USDA-ARS-Animal Physiology Research Unit Athens, GA 30605 706-583-8279 706-542-0399 (fax) ------------------------------ Message: 14 Date: Tue, 05 Oct 2004 17:44:50 -0400 From: "Nicole Hedgecock" Subject: [Histonet] More details: decalcified sections of bone To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=iso-8859-1; format=flowed I sent this out earlier (see below) and more details of my tissue processing and sectioning were requested. I want to identify apoptotic osteocytes in rabbit cortical bone as well as some morphological features especially microcracks. I fix the bones in 4% paraformaldehyde for 24 hours, then place then in 70% EtOH until I can decalcify them in EDTA at room temp. Between the EtOH and EDTA steps I rinse the bones in dH2O for about 3 days. After decalcification, the bones are again rinsed in dH2O and then embedded in paraffin and sectioned to 6 microns. I use the TUNEL kit from Roche for In situ labeling of DNA breaks. Do you think that I have to do decalcification for the TUNEL to work? Does sectioning of paraffin embedded tissue create a lot of microscopic damage to the section? I hope this clears up my situation and goals. Nicole >From: "Nicole Hedgecock" >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] decalcified sections of bone >Date: Tue, 05 Oct 2004 14:42:45 -0400 > >Hi, All > >I am attempting to do TUNEL staining of osteocytes in cortical bone. I was >wondering why the bone has to be decalcified in order to do this process. >And Why do the sections have to be so thin? > >Thanks >Nicole > > > >Nicole Hedgecock >Master's Student >Orthopedic Research Lab >UCD Medical Center >University of California, Davis > >_________________________________________________________________ >?Cu?nto vale tu auto? Tips para mantener tu carro. ?De todo en MSN Latino >Autos! http://latino.msn.com/autos/ > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Visita MSN Latino Entretenimiento: ?m?sica, cine, chismes, TV y m?s...! http://latino.msn.com/entretenimiento/ ------------------------------ Message: 15 Date: Tue, 5 Oct 2004 16:53:39 -0500 From: lkbauer@unmc.edu Subject: [Histonet] orienting mouse embryos To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Could I tap your collective experience to help solve a problem? I am trying to orient gestational day 9.5 mouse embryos in paraffin for cross sectioning. After processing, these specimens are like pieces of fragil white pepper. I can only see head from tail with a microscope. By the time I get everything in focus the parffin is turning white....like finding a ghost in a blizzard. Does anyone have any tips for this task? Thanks, Lin Linda(Lin)Bauer Department of Genetics, Cell Biology, and Anatomy University of Nebraska Medical Center Omaha, NE 68198-5455 Email: lkbauer@unmc.edu ------------------------------ Message: 16 Date: Tue, 5 Oct 2004 18:10:01 -0400 From: "Bonner, Janet" Subject: RE: [Histonet] orienting mouse embryos To: "'lkbauer@unmc.edu'" , histonet@lists.utsouthwestern.edu Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4092@fh2k093.fhmis.net> Content-Type: text/plain; charset="iso-8859-1" Put eosin-phloxine stain (80ml to 3500ml) in your first alcohol when processing, we use this on our biopsies and it makes all the difference!. Janet -----Original Message----- From: lkbauer@unmc.edu [mailto:lkbauer@unmc.edu] Sent: Tuesday, October 05, 2004 5:54 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] orienting mouse embryos Could I tap your collective experience to help solve a problem? I am trying to orient gestational day 9.5 mouse embryos in paraffin for cross sectioning. After processing, these specimens are like pieces of fragil white pepper. I can only see head from tail with a microscope. By the time I get everything in focus the parffin is turning white....like finding a ghost in a blizzard. Does anyone have any tips for this task? Thanks, Lin Linda(Lin)Bauer Department of Genetics, Cell Biology, and Anatomy University of Nebraska Medical Center Omaha, NE 68198-5455 Email: lkbauer@unmc.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. ------------------------------ Message: 17 Date: Wed, 06 Oct 2004 08:18:08 +1000 From: Laurie Reilly Subject: Re: [Histonet] orienting mouse embryos To: lkbauer@unmc.edu, histonet@lists.utsouthwestern.edu Message-ID: <5.1.0.14.0.20041006081451.00a39b30@mail.jcu.edu.au> Content-Type: text/plain; charset="us-ascii"; format=flowed Dear Lin, Try putting a few mls of aqueous eosin in your fixative. A pink specimen is a lot easier to see. The eosin can be washed out at a later stage if it is not required. Good luck and regards, Laurie. At 04:53 PM 10/05/04 -0500, lkbauer@unmc.edu wrote: >Could I tap your collective experience to help solve a problem? I am trying >to orient gestational day 9.5 mouse embryos in paraffin for cross >sectioning. After processing, these specimens are like pieces of fragil >white pepper. I can only see head from tail with a microscope. By the time >I get everything in focus the parffin is turning white....like finding a >ghost in a blizzard. Does anyone have any tips for this task? Thanks, Lin > >Linda(Lin)Bauer >Department of Genetics, Cell Biology, and Anatomy >University of Nebraska Medical Center >Omaha, NE 68198-5455 >Email: lkbauer@unmc.edu > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mr.Laurie Reilly Ph 07 4781 4468 Physiology & Pharmacology Fax 07 4779 1526 Aust.Inst.of Tropical Vet.& Animal Sc. James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. ------------------------------ Message: 18 Date: Tue, 5 Oct 2004 17:15:33 -0500 From: ajennings@unmc.edu Subject: Re: [Histonet] orienting mouse embryos To: lkbauer@unmc.edu Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Lin Run downstairs and I will show you the tricks of the trade.....thought I had already shown you :) ------------------------------ Message: 19 Date: Tue, 5 Oct 2004 19:01:18 EDT From: Gervaip@aol.com Subject: [Histonet] Reusing the filter for ThinPrep for HPV To: histonet@pathology.swmed.edu Message-ID: <110.398ec1e3.2e9481be@aol.com> Content-Type: text/plain; charset="US-ASCII" Is anyone in HistoLand doing this? We were told we can reuse the filter to prepare another slide for HPV. If you do, how do you dry the filter? Pearl in New Orleans ------------------------------ Message: 20 Date: Wed, 6 Oct 2004 12:51:05 +1300 From: "Darren James" Subject: [Histonet] Bone Saw and Dust Extraction To: "Histonet \(E-mail\)" Message-ID: <000e01c4ab36$2e3140e0$c864a8c0@medica.co.nz> Content-Type: text/plain; charset="iso-8859-1" Hello, I have a question, does anyone know if there is an electric bone saw available complete with a dust extraction unit for a Histology lab? If not, would a saw with a water spray eliminate aerosol and particulate matter? I am looking for a benchtop saw for a histo lab not a Stryker or similar mortuary saw. Thanks Darren James Technical Representative Medica Pacifica Ltd Office Address: 1A 153 Stoddard Road, Mt Roskill, Auckland Postal Address: PO Box 24-421, Royal Oak, Auckland 1030 Ph: +64 9 629 0823 Fax: +64 9 629 0542 Mob: + 64 21 633 820 ------------------------------ Message: 21 Date: Tue, 5 Oct 2004 23:01:49 -0400 (EDT) From: "SMITH,REBEKAH FELICIA" Subject: Re: [Histonet] DAB-colors... To: "Louro, Pedro" , Histonet@lists.utsouthwestern.edu Message-ID: <119304449.1097031709998.JavaMail.osg@osgjas02.cns.ufl.edu> Content-Type: text/plain; format=flowed; charset=us-ascii The only two colors I know of for DAB are brown and black. The black is due to adding nickel to DAB, I think. I think pierce sells them. On Tue Oct 05 10:48:28 EDT 2004, "Louro, Pedro" wrote: > Hello all, > > When at NSH -Toronto..I heard people talking about using DAB in > different > colors. > Can someone tell who sells this product. > > Thanks Pedro > > > ********************************************************************* > This message and any attachments are solely for the intended > recipient. If you are not the intended recipient, disclosure, > copying, use or distribution of the information included in this > message is prohibited -- Please immediately and permanently > delete. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" ------------------------------ Message: 22 Date: Wed, 6 Oct 2004 08:46:37 +0200 (CEST) From: Jose Luis Palazon Fernandez Subject: [Histonet] elastic fibers To: Histonet@lists.utsouthwestern.edu Message-ID: <20041006064637.2BF5831C921@perceval.uca.es> Content-Type: text/plain; charset="iso-8859-1" Dear List fellows Many thanks for all of you who answered to my question on elastic fibers stain. As I can see the more popular stains are in order of preference 1)Verhoeff's Van Gieson 2) Weigher resorcin fuchsin. Greetings Jos? Luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es ------------------------------ Message: 23 Date: Wed, 6 Oct 2004 06:16:45 -0500 From: "Molinari, Betsy" Subject: [Histonet] processing question To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hi histonetters, Is it harmful to tissue that is in 70% ETOH to be put on the processor and start at 10% NBF? Thanks . Betsy Molinari HT(ASCP) Texas Heart Institute Houston,TX 77030 832-355-6524 ------------------------------ Message: 24 Date: Wed, 6 Oct 2004 08:18:46 -0500 From: "Joe Nocito" Subject: RE: [Histonet] Reusing the filter for ThinPrep for HPV To: , Message-ID: Content-Type: text/plain; charset="us-ascii" Pearl, we routinely put the patient's filter in the Preservecyte vial after the Thin prep is made. If we need o go back to that vial, we empty whatever material is in the filter, back into the vial. Very carefully, we wipe both sides of the filter with a Kimwipe and let the filter air dry (about a minute. We try not to put too much pressure on the filter because sometimes it comes off. Depending on the cellularity of the specimen, I have been able to get 3 additional slides before I had to get another filter. I think they clog up after a while. Now, if anyone from Cytyc is listening, my rep is Billy Redding. Disclaimer: This method is not approved by Cytyc, the FDA, the FCC, the FBI and the CIA, the GOP, nor the DNC. I will be expecting a phone call from Billy any minute now. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gervaip@aol.com Sent: Tuesday, October 05, 2004 6:01 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Reusing the filter for ThinPrep for HPV Is anyone in HistoLand doing this? We were told we can reuse the filter to prepare another slide for HPV. If you do, how do you dry the filter? Pearl in New Orleans _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 25 Date: Wed, 6 Oct 2004 06:49:19 -0700 From: Paula Lucas Subject: [Histonet] Cassette Label System To: "Histonet (E-mail)" Message-ID: <01C4AB70.9B2AB250.plucas@biopath.org> Hello, My company is possibly interested in purchasing a histology cassette printer. Could you please provide information on the different manufactures that offer printers? What are the pro/cons? Is it really that worth it to purchase one? Does it really save that much time? Are they easy to use? Any information, suggestions, and comments are greatly appreciated. Thank you, Paula Lucas Bio-Path Medical Group ------------------------------ Message: 26 Date: Wed, 06 Oct 2004 08:48:23 -0500 From: "Jordan Brod" Subject: [Histonet] Eye Protection Question To: Message-ID: Content-Type: text/plain; charset=US-ASCII Good Morning! What kind of eye protection are technicians required to wear in the laboratory during grossing, sectioning, processor maintenance, etc.? Thanks and have a great day. Jordan TVMDL - College Station, TX ------------------------------ Message: 27 Date: Wed, 6 Oct 2004 09:22:40 -0500 From: Jackie.O'Connor@abbott.com Subject: Re: [Histonet] Eye Protection Question To: "Jordan Brod" Cc: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Protection from biohaz splash where appropriate, protection from chemical splash, where appropriate. As a note - just to walk into our labs here, even just to say hi to another person, we must have our safety glasses and labcoats on. The thinking is that someone else working may create a splash that sails across the room into your eye. "Jordan Brod" Sent by: histonet-bounces@lists.utsouthwestern.edu 10/06/2004 08:48 AM To: cc: Subject: [Histonet] Eye Protection Question Good Morning! What kind of eye protection are technicians required to wear in the laboratory during grossing, sectioning, processor maintenance, etc.? Thanks and have a great day. Jordan TVMDL - College Station, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 28 Date: Wed, 6 Oct 2004 10:55:37 -0500 From: "Joe Nocito" Subject: RE: [Histonet] Cassette Label System To: "Paula Lucas" , "Histonet \(E-mail\)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Paula, we just purchased a cassette printer from Surgipath. It prints one block at a time, but can be set to automatically increment. We tested one printer from TBS, but the cassettes kept jamming. The Surgipath printer maybe slower, but it has less moving parts to break down and it does a great job. Disclaimer: As usual, the opinions of the author in no way reflect the opinions of the company, it executives or relatives. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Paula Lucas Sent: Wednesday, October 06, 2004 8:49 AM To: Histonet (E-mail) Subject: [Histonet] Cassette Label System Hello, My company is possibly interested in purchasing a histology cassette printer. Could you please provide information on the different manufactures that offer printers? What are the pro/cons? Is it really that worth it to purchase one? Does it really save that much time? Are they easy to use? Any information, suggestions, and comments are greatly appreciated. Thank you, Paula Lucas Bio-Path Medical Group _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 29 Date: Wed, 6 Oct 2004 10:59:39 -0500 From: "Joe Nocito" Subject: RE: [Histonet] Eye Protection Question To: "Jordan Brod" , Message-ID: Content-Type: text/plain; charset="US-ASCII" Jordon, we have full-face safety shields as well as surgical masks with an attached eye shield. We wear either when changing the staining machines and tissue processors. When I gross, I wear my eyeglasses that are classified as safety wear. We have never had an incident, but we flush our eyewash stations weekly because you never know what will happen. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jordan Brod Sent: Wednesday, October 06, 2004 8:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Eye Protection Question Good Morning! What kind of eye protection are technicians required to wear in the laboratory during grossing, sectioning, processor maintenance, etc.? Thanks and have a great day. Jordan TVMDL - College Station, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 30 Date: Wed, 6 Oct 2004 12:40:36 -0400 From: "Fawn Jones" Subject: [Histonet] micronucleus staining To: Message-ID: <915E55B02E236E4D95258B181EEF6317075D2B@namsams01.namsa.int> Content-Type: text/plain; charset="us-ascii" Hi everyone, Our histology labs is doing a wright-giemsa stain on mice micronucleus slides and we are having a problem with our cells not picking up the stain. Are there any suggestions for us. We let them dry overnight then we put them in Methanol for 5 minutes, let them air dry for 20 minutes then they go into Wright's stain for 6 minutes, then Wright-giemsa stain for 6 minutes, and then go through 3 changes of sterile water for injection for 30 seconds each. Then they sit overnight before coverslipping. If anyone could help that would be great. Thanks Fawn Jones North American Science Associates ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 11, Issue 7 *************************************** From rkrug <@t> sial.com Wed Oct 6 12:26:50 2004 From: rkrug <@t> sial.com (Robert Krug) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] micronucleus staining Message-ID: Fawn: The sterile water you are using is too acidic for the staining you are performing. Bottle distilled water often has a pH around 4.0 to 4.5 Sterile water (if it has sat for any period of time) is probably as acidic as standard bottle distilled water. In any Romanowsky type staining procedure (of which Wright Giemsa is included) the water is actually a differentiator. With human peripheral blood or bone marrow cells, the proper pH is typically 6.8 to 7.2. Some animals require slightly lower pH. Paraffin processed tissue sections for some reason I don't understand appear to accept lower pH values. With the pH of the sterile water you are using, the color which is present in the cells is being stripped away by the acidic water you are using. Regardless of how well your Wright Stain solution may be working, exposing the slides to 3 changes of acidic water is more than likely stripping any color from the slides. The latest edition of Humason's Animal Tissues Techniques, 5th edition has the formulations for Sorensen's phosphate buffer listed on pages 473 and 474. Sigma-Aldrich sells a slightly lower molarity product as P3288. Try an appropriate pH phosphate or TRIS buffer and I think you will find your staining improves. I would also suggest you fix your slides after 30 minutes of drying in the methanol. Waiting overnight before fixation allows for oxidation of the hemoglobin. Best staining results occur if the blood is stained right after collection. You may also want to cut back on your staining times in Wright Stain and Wright Giemsa solutions. The original Wright Giemsa procedure had the user first staining in Wright Stain and then following up with a Giemsa procedure. I do not quite understand the concept of first staining in Wright Stain and then staining a second time in Wright Giemsa stain. More typical staining times for blood smears would range from about 15 seconds to 2 minutes for the Wright Stain and typically double the amount of time in deionized water or buffer. The 1 to 2 ratio is a good starting point for the procedure, but the times can be adjusted to suit personal color preference. Wright Giemsa or Giemsa solutions typically require longer incubations and are more commonly performed on bone marrow preparations sor cultured cells. Giemsa is used for identifying malarial parasites and for tissue sections. If you want to bring out more red in your red blood cells, try skipping the Wright Giemsa stain and use just the Wright Stain. Best Regards, Bob Krug Technical Marketing Specialist Sigma-Aldrich St Louis, Missouri e-mail:? clintech@sial.com (800) 325-0250 Order your new MedBasics Product Guide today-laboratory essentials for medical testing. Call 800-282-1298 or click here to reserve your copy? www.sigmaaldrich.com/medbasics "Fawn Jones" Sent by: histonet-bounces@lists.utsouthwestern.edu 10/06/2004 11:40 AM To: cc: Subject: [Histonet] micronucleus staining Hi everyone, Our histology labs is doing a wright-giemsa stain on mice micronucleus slides and we are having a problem with our cells not picking up the stain. Are there any suggestions for us. We let them dry overnight then we put them in Methanol for 5 minutes, let them air dry for 20 minutes then they go into Wright's stain for 6 minutes, then Wright-giemsa stain for 6 minutes, and then go through 3 changes of sterile water for injection for 30 seconds each. Then they sit overnight before coverslipping. If anyone could help that would be great. Thanks Fawn Jones North American Science Associates _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Wed Oct 6 12:36:18 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] thanks for answering IHC-dewaxing-question Message-ID: <002d01c4abcb$035e02a0$eeeea8c0@server> From lizchlipala <@t> premierhistology.com Wed Oct 6 12:57:08 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] fluorescent staining of nuclei or nuclear material Message-ID: <000601c4abcd$e6255c30$76d48a80@AMY> Hello everyone I have a client that has some tissue (fascia) samples which have been processed in an attempt to remove the intrinsic cells. They have some H&E sections, but want to use a more sensitive technique to ensure that no cell remnants or nuclear material remains. Is there is a fluorescent staining method to visualize cell nuclear material. I have gone to the molecular probes web page and there is a wide variety of nuclear stains available. Does anyone have some advice on what would be the best method for this application and whether or not it would work on paraffin embedded material. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Histology Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 lizchlipala@premierhistology.com www.premierhistology.com Ship to Address: Premier Histology Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From gcallis <@t> montana.edu Wed Oct 6 13:19:10 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] DAPI for fluorescent staining of DNAl In-Reply-To: <000601c4abcd$e6255c30$76d48a80@AMY> References: <000601c4abcd$e6255c30$76d48a80@AMY> Message-ID: <6.0.0.22.1.20041006121417.01b56e38@gemini.msu.montana.edu> Try DAPI, and you can buy mounting media from Vector called Vectashield that contains DAPI. You will have to have the correct filter in order to view DAPI, but VECTOR can provide that info., also Molecular Probes. You can talk to Craig Pow at Vector about specs with DAPI, but it should work and is an aqueous mounting media, rehydrate sections to water, and coverslip. You can contact Craig at cspow@vectorlabs.com if you have questions as to whether this works with FFPE. DAPI binds to DNA and fluoresces a pretty bluish/white color. At 11:57 AM 10/6/2004, you wrote: >Hello everyone > >I have a client that has some tissue (fascia) samples which have been >processed in an attempt to remove the intrinsic cells. They have some >H&E sections, but want to use a more sensitive technique to ensure that >no cell remnants or nuclear material remains. Is there is a fluorescent >staining method to visualize cell nuclear material. I have gone to the >molecular probes web page and there is a wide variety of nuclear stains >available. Does anyone have some advice on what would be the best >method for this application and whether or not it would work on paraffin >embedded material. > >Thanks in advance. > >Liz > >Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC >Premier Histology Laboratory, LLC >P.O. Box 18592 >Boulder, Colorado 80308 >Office: (303) 735-5001 >Fax: (303) 735-3540 >lizchlipala@premierhistology.com >www.premierhistology.com > >Ship to Address: >Premier Histology Laboratory >University of Colorado >MCDB, Room A3B40 >Boulder, Colorado 80309 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From jkiernan <@t> uwo.ca Wed Oct 6 13:52:44 2004 From: jkiernan <@t> uwo.ca (J. A. Kiernan) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] fluorescent staining of nuclei or nuclear material References: <000601c4abcd$e6255c30$76d48a80@AMY> Message-ID: <41643EFC.7404B29E@uwo.ca> Fluorescence microscopy is useful for seeing small numbers of bright objects against a dark background. An ordinary Feulgen stain, though done for ordinary microscopy, is also fluorescent. It is specific for DNA if you do the proper controls. Any fluorescent yellow cationic dye will stain nuclei. Acriflavine (about 0.001%) is cheap and effective. Bisbenzimide (also called Hoechst 33258) is more often used; it can be incorporated into an aqueous mounting medium. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Elizabeth Chlipala wrote: > > Hello everyone > > I have a client that has some tissue (fascia) samples which have been > processed in an attempt to remove the intrinsic cells. They have some > H&E sections, but want to use a more sensitive technique to ensure that > no cell remnants or nuclear material remains. Is there is a fluorescent > staining method to visualize cell nuclear material. I have gone to the > molecular probes web page and there is a wide variety of nuclear stains > available. Does anyone have some advice on what would be the best > method for this application and whether or not it would work on paraffin > embedded material. > > Thanks in advance. > > Liz > > Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC > Premier Histology Laboratory, LLC > P.O. Box 18592 > Boulder, Colorado 80308 > Office: (303) 735-5001 > Fax: (303) 735-3540 > lizchlipala@premierhistology.com > www.premierhistology.com > > Ship to Address: > Premier Histology Laboratory > University of Colorado > MCDB, Room A3B40 > Boulder, Colorado 80309 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcline <@t> wchsys.org Wed Oct 6 13:54:03 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] PROSTATE SEEDS Message-ID: <000001c4abd5$d93799e0$1d2a14ac@wchsys.org> Can someone share with me the info on receiving a specimen with prostate seeds? I already have a procedure for sentinel nodes but seems I need one for the unlikely chance I may receive prostate tissue with radiation seeds. Thanks Joyce ***** CONFIDENTIALITY NOTICE ***** This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. From flemons <@t> bhset.org Wed Oct 6 14:00:24 2004 From: flemons <@t> bhset.org (Fran Lemons) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Results Message-ID: Hello fellow netters...... Can anyone give me an educated guess as to when we can expect results for the practical exam for this cycle? I have 2 techs on pins & needles waiting to hear. They already have results for the written, but we were wondering how long before the practical gets graded & we hear anything. Thanks Fran Walker HTS Baptist Hospital of E. TN (Knoxville) From gentras <@t> vetmed.auburn.edu Wed Oct 6 14:08:50 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] mouse brains Message-ID: <6.0.1.1.0.20041006140230.025c7370@mailhost.vetmed.auburn.edu> Hello, those of you processing & sectioning coronal sections of mouse brain; from you experience can prolonged fixation in 4%PFA be the culprit of sectioning problems? What is the maximum fixation time period the brain should held in PFA before processing & sectioning? Thanks! Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From darrenj <@t> medica.co.nz Wed Oct 6 14:42:24 2004 From: darrenj <@t> medica.co.nz (Darren James) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Reusing the filter for ThinPrep for HPV In-Reply-To: Message-ID: <000501c4abdc$9af02240$c864a8c0@medica.co.nz> I have also been told that you can reuse the Thin Prep filter in exactly the way Joe mentions. It was also mentioned that you can "back flush" the filter by gently pouring thin prep fixative back into it to prevent clogging. I have never done this so don't know if it really works or merely anecdotal Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joe Nocito Sent: Thursday, 7 October 2004 2:19 a.m. To: Gervaip@aol.com; histonet@pathology.swmed.edu Subject: RE: [Histonet] Reusing the filter for ThinPrep for HPV Pearl, we routinely put the patient's filter in the Preservecyte vial after the Thin prep is made. If we need o go back to that vial, we empty whatever material is in the filter, back into the vial. Very carefully, we wipe both sides of the filter with a Kimwipe and let the filter air dry (about a minute. We try not to put too much pressure on the filter because sometimes it comes off. Depending on the cellularity of the specimen, I have been able to get 3 additional slides before I had to get another filter. I think they clog up after a while. Now, if anyone from Cytyc is listening, my rep is Billy Redding. Disclaimer: This method is not approved by Cytyc, the FDA, the FCC, the FBI and the CIA, the GOP, nor the DNC. I will be expecting a phone call from Billy any minute now. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gervaip@aol.com Sent: Tuesday, October 05, 2004 6:01 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Reusing the filter for ThinPrep for HPV Is anyone in HistoLand doing this? We were told we can reuse the filter to prepare another slide for HPV. If you do, how do you dry the filter? Pearl in New Orleans _______________________________________________ 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 From darrenj <@t> medica.co.nz Wed Oct 6 14:50:58 2004 From: darrenj <@t> medica.co.nz (Darren James) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Cassette Label System In-Reply-To: <01C4AB70.9B2AB250.plucas@biopath.org> Message-ID: <000601c4abdd$ccd4b270$c864a8c0@medica.co.nz> Hi Paula, As agents for Thermo Electron in New Zealand I am going to extoll the virutes of the Shandon models. Unlike some on the market the Shandon uses foil tape and heated stylus. This burns the info into the cassette. They can print conventional and 2D barcodes and manage to print up to 3 lines of 19 characters. The software is fully configurable to customise the printing to your requirements. I have contacted several users througout NZ and Australia who all love the product. A couple of labs have 2 or 3 running at the same time. I think the biggest benefit is having totally legible cassettes and therefore a clear audit trail. Other users may like to comment. To get more info I suggest contacting your Thermo Electron agent or email them on clinical.diagnostics@thermo.com. Hope this helps Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Paula Lucas Sent: Thursday, 7 October 2004 2:49 a.m. To: Histonet (E-mail) Subject: [Histonet] Cassette Label System Hello, My company is possibly interested in purchasing a histology cassette printer. Could you please provide information on the different manufactures that offer printers? What are the pro/cons? Is it really that worth it to purchase one? Does it really save that much time? Are they easy to use? Any information, suggestions, and comments are greatly appreciated. Thank you, Paula Lucas Bio-Path Medical Group _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From darrenj <@t> medica.co.nz Wed Oct 6 14:58:15 2004 From: darrenj <@t> medica.co.nz (Darren James) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Eye Protection Question In-Reply-To: Message-ID: <000c01c4abde$d1f94440$c864a8c0@medica.co.nz> Hi, I agree totally with Joe's point about flushing your eyewash stations regularly. These are often forgotten about and can become incredibly contaminated over a very short period. We had micro prepare some plates on the water in the eyewash stations and there were some fairly pathogenic species present. Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joe Nocito Sent: Thursday, 7 October 2004 5:00 a.m. To: Jordan Brod; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Eye Protection Question Jordon, we have full-face safety shields as well as surgical masks with an attached eye shield. We wear either when changing the staining machines and tissue processors. When I gross, I wear my eyeglasses that are classified as safety wear. We have never had an incident, but we flush our eyewash stations weekly because you never know what will happen. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jordan Brod Sent: Wednesday, October 06, 2004 8:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Eye Protection Question Good Morning! What kind of eye protection are technicians required to wear in the laboratory during grossing, sectioning, processor maintenance, etc.? Thanks and have a great day. Jordan TVMDL - College Station, TX _______________________________________________ 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 From histopath <@t> cvm.tamu.edu Wed Oct 6 14:58:23 2004 From: histopath <@t> cvm.tamu.edu (Histopath) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] antibody organizers Message-ID: <1453BDD1-17D2-11D9-8A9D-0005029D8A73@cvm.tamu.edu> Does anyone know of anyone that sells organizers/storage for stock antibody vials? If anyone has a creative solution that would be great also. Thanks Susan Hernandez From MAUGER <@t> email.chop.edu Wed Oct 6 15:19:14 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] mouse brains Message-ID: To Atoska, My experience shows that PFA isn't good enough as a fixative. I fix the tissue again in 10% NBF for 6-12 hours, and process as usual. Try it. It really is much better. Jo >>> "Atoska S. Gentry" 10/06/04 03:08PM >>> Hello, those of you processing & sectioning coronal sections of mouse brain; from you experience can prolonged fixation in 4%PFA be the culprit of sectioning problems? What is the maximum fixation time period the brain should held in PFA before processing & sectioning? Thanks! Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mbecker <@t> pathlabinc.com Wed Oct 6 15:21:10 2004 From: mbecker <@t> pathlabinc.com (Michele Becker) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] GI bx Message-ID: Please help.................... We are having problems with blurring and distorted nuclei on some of our GI biopsies. This has been happening on and off for the past few months. It is not every day and not all the GI bx on any given day. We have ruled out staining and processing. I really think it is something happening to the bxs before we even receive them but want to make sure we have not left any stones unturned on our end first. Any suggestions? Thank-you. Michele From darrenj <@t> medica.co.nz Wed Oct 6 15:34:35 2004 From: darrenj <@t> medica.co.nz (Darren James) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] antibody organizers In-Reply-To: <1453BDD1-17D2-11D9-8A9D-0005029D8A73@cvm.tamu.edu> Message-ID: <000e01c4abe3$e53247a0$c864a8c0@medica.co.nz> Hi Susan, Check out a company called Heathrow Scientific www.heathrowscientific.com They have some very useful plastic rack and storage systems. Thanks Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Histopath Sent: Thursday, 7 October 2004 8:58 a.m. To: histonet@pathology.swmed.edu Subject: [Histonet] antibody organizers Does anyone know of anyone that sells organizers/storage for stock antibody vials? If anyone has a creative solution that would be great also. Thanks Susan Hernandez _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RossS <@t> BaylorHealth.edu Wed Oct 6 15:36:04 2004 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] antibody organizers Message-ID: At a former lab, I filled a flat tupperware type container with paraffin and as it solidified on the cold plate of the embedding center I pushed an empty vial down into the paraffin to create the proper sized hole. I just kept punching out holes in the paraffin in rows until I had as many as the container would allow. Then you have the option of removing the paraffin from the container and re-using the container as a mold for more paraffin racks or leaving it in the container so that it looks a little neater. The drawback is that your "vial rack" is a bit heavy. The one I made lasted a few years. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histopath Sent: Wednesday, October 06, 2004 2:58 PM To: histonet@pathology.swmed.edu Subject: [Histonet] antibody organizers Does anyone know of anyone that sells organizers/storage for stock antibody vials? If anyone has a creative solution that would be great also. Thanks Susan Hernandez _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Wed Oct 6 15:53:32 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] mouse brains Message-ID: Are you perfusing, or just extracting and dropping in PFA? Sorry, what is PFA? Perfusion is necessary for good fixation, and is easy to do. Overfixation can lead to loss of immuno or other reactivity, but can only improve sectioning. I would guess you are not fixing fast enough, your tissue is bad before fixative gets a chance to do its work. Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Atoska S. Gentry Sent: Wednesday, October 06, 2004 2:09 PM To: Histonet Subject: [Histonet] mouse brains Hello, those of you processing & sectioning coronal sections of mouse brain; from you experience can prolonged fixation in 4%PFA be the culprit of sectioning problems? What is the maximum fixation time period the brain should held in PFA before processing & sectioning? Thanks! Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cforster <@t> umn.edu Wed Oct 6 16:26:19 2004 From: cforster <@t> umn.edu (Colleen Forster) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] 4% PFA Message-ID: <416462FB.1040409@umn.edu> 4% PFA is a very good fixative. The difference between 4% and 10% is that the PFA is made fresh each time and has no impurities added ex: methanols. The formalin content in both comes out to about the same. It is a standard fixative for many research labs. You can drop fix with PFA or perfuse and then drop fix for additional time for very good fixation and excellent results. I have used it for my mouse brain tissue for over 6 years with very nice results. Colleen Forster U of Mn From JMyers1 <@t> aol.com Wed Oct 6 17:01:31 2004 From: JMyers1 <@t> aol.com (JMyers1@aol.com) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Re: Antibody organizers Message-ID: Personally, I use the compartmental organizers designed for fishing tackle, with hinged lids, that are available at most discount and sporting goods stores. Alternatively, one could a 'divider box' like those offered by MarketLab (www. marketlabinc. com). J.D. Myers --------------------------------------------------------- Message: 14 Date: Wed, 6 Oct 2004 14:58:23 -0500 From: Histopath Subject: [Histonet] antibody organizers Does anyone know of anyone that sells organizers/storage for stock antibody vials? If anyone has a creative solution that would be great also. Thanks Susan Hernandez From Evelyn.Flynn <@t> childrens.harvard.edu Wed Oct 6 17:09:14 2004 From: Evelyn.Flynn <@t> childrens.harvard.edu (Flynn, Evelyn) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] antibody organizers Message-ID: Dear Susan, In my previous lab I used a plastic "hardware organizer" with a dozen or more small drawers that a person might use for nails, nuts, and bolts. Usually one drawer was designated and labelled for the vials from one or two companies. I believe it came from a hardware store. Good luck, Evelyn Flynn -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Histopath Sent: Wed 10/6/2004 3:58 PM To: histonet@pathology.swmed.edu Subject: [Histonet] antibody organizers Does anyone know of anyone that sells organizers/storage for stock antibody vials? If anyone has a creative solution that would be great also. Thanks Susan Hernandez _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JQB7 <@t> CDC.GOV Wed Oct 6 17:22:05 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Eye Protection Question Message-ID: We flush ours weekly as well. We cover them with a small plastic garbage bag and let them run a minimum of 10 minutes. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Darren James Sent: Wed 10/6/2004 3:58 PM To: 'Joe Nocito'; Histonet (E-mail) Cc: Subject: RE: [Histonet] Eye Protection Question Hi, I agree totally with Joe's point about flushing your eyewash stations regularly. These are often forgotten about and can become incredibly contaminated over a very short period. We had micro prepare some plates on the water in the eyewash stations and there were some fairly pathogenic species present. Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joe Nocito Sent: Thursday, 7 October 2004 5:00 a.m. To: Jordan Brod; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Eye Protection Question Jordon, we have full-face safety shields as well as surgical masks with an attached eye shield. We wear either when changing the staining machines and tissue processors. When I gross, I wear my eyeglasses that are classified as safety wear. We have never had an incident, but we flush our eyewash stations weekly because you never know what will happen. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jordan Brod Sent: Wednesday, October 06, 2004 8:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Eye Protection Question Good Morning! What kind of eye protection are technicians required to wear in the laboratory during grossing, sectioning, processor maintenance, etc.? Thanks and have a great day. Jordan TVMDL - College Station, TX _______________________________________________ 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 From JQB7 <@t> CDC.GOV Wed Oct 6 17:23:12 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Cassette Label System Message-ID: I saw the ThermoElectron unit demo'd at NSH a few weeks ago. It looked really amazing. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Darren James Sent: Wed 10/6/2004 3:50 PM To: 'Paula Lucas'; Histonet (E-mail) Cc: Subject: RE: [Histonet] Cassette Label System Hi Paula, As agents for Thermo Electron in New Zealand I am going to extoll the virutes of the Shandon models. Unlike some on the market the Shandon uses foil tape and heated stylus. This burns the info into the cassette. They can print conventional and 2D barcodes and manage to print up to 3 lines of 19 characters. The software is fully configurable to customise the printing to your requirements. I have contacted several users througout NZ and Australia who all love the product. A couple of labs have 2 or 3 running at the same time. I think the biggest benefit is having totally legible cassettes and therefore a clear audit trail. Other users may like to comment. To get more info I suggest contacting your Thermo Electron agent or email them on clinical.diagnostics@thermo.com. Hope this helps Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Paula Lucas Sent: Thursday, 7 October 2004 2:49 a.m. To: Histonet (E-mail) Subject: [Histonet] Cassette Label System Hello, My company is possibly interested in purchasing a histology cassette printer. Could you please provide information on the different manufactures that offer printers? What are the pro/cons? Is it really that worth it to purchase one? Does it really save that much time? Are they easy to use? Any information, suggestions, and comments are greatly appreciated. Thank you, Paula Lucas Bio-Path Medical Group _______________________________________________ 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 From jwatson <@t> gnf.org Wed Oct 6 17:32:44 2004 From: jwatson <@t> gnf.org (James Watson) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Cassette Label System Message-ID: Recently purchased a thermo Electron system and it is working very well. James Watson HT, ASCP Facilities Manager of Histology GNF, C015 858-332-4647 jwatson@gnf.org -----Original Message----- From: Bartlett, Jeanine [mailto:JQB7@CDC.GOV] Sent: Wednesday, October 06, 2004 3:23 PM To: darrenj@medica.co.nz; Paula Lucas; Histonet (E-mail) Subject: RE: [Histonet] Cassette Label System I saw the ThermoElectron unit demo'd at NSH a few weeks ago. It looked really amazing. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Darren James Sent: Wed 10/6/2004 3:50 PM To: 'Paula Lucas'; Histonet (E-mail) Cc: Subject: RE: [Histonet] Cassette Label System Hi Paula, As agents for Thermo Electron in New Zealand I am going to extoll the virutes of the Shandon models. Unlike some on the market the Shandon uses foil tape and heated stylus. This burns the info into the cassette. They can print conventional and 2D barcodes and manage to print up to 3 lines of 19 characters. The software is fully configurable to customise the printing to your requirements. I have contacted several users througout NZ and Australia who all love the product. A couple of labs have 2 or 3 running at the same time. I think the biggest benefit is having totally legible cassettes and therefore a clear audit trail. Other users may like to comment. To get more info I suggest contacting your Thermo Electron agent or email them on clinical.diagnostics@thermo.com. Hope this helps Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Paula Lucas Sent: Thursday, 7 October 2004 2:49 a.m. To: Histonet (E-mail) Subject: [Histonet] Cassette Label System Hello, My company is possibly interested in purchasing a histology cassette printer. Could you please provide information on the different manufactures that offer printers? What are the pro/cons? Is it really that worth it to purchase one? Does it really save that much time? Are they easy to use? Any information, suggestions, and comments are greatly appreciated. Thank you, Paula Lucas Bio-Path Medical Group _______________________________________________ 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 From victor <@t> pathology.washington.edu Wed Oct 6 17:34:26 2004 From: victor <@t> pathology.washington.edu (Victor Tobias) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Cassette Label System References: Message-ID: <416472F2.4090801@pathology.washington.edu> We have 2 ThermoShandon microwriter cassette labelers and knock on wood, have not had any downtime. One is 3 years old and the other 18 months. We average 300 + cassettes daily. We are upgrading our labelers so that we can print barcodes. Our labelers are also integrated with our LIS, PowerPath by IMPAC. As soon as the case is accessioned and block orders are placed, upon saving the cassettes are automatically printed and color coded. Victor Bartlett, Jeanine wrote: >I saw the ThermoElectron unit demo'd at NSH a few weeks ago. It looked really amazing. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Darren James > Sent: Wed 10/6/2004 3:50 PM > To: 'Paula Lucas'; Histonet (E-mail) > Cc: > Subject: RE: [Histonet] Cassette Label System > > > > Hi Paula, > > As agents for Thermo Electron in New Zealand I am going to extoll the > virutes of the Shandon models. > Unlike some on the market the Shandon uses foil tape and heated stylus. This > burns the info into the cassette. > They can print conventional and 2D barcodes and manage to print up to 3 > lines of 19 characters. The software is fully configurable to customise the > printing to your requirements. > I have contacted several users througout NZ and Australia who all love the > product. A couple of labs have 2 or 3 running at the same time. > I think the biggest benefit is having totally legible cassettes and > therefore a clear audit trail. > Other users may like to comment. > To get more info I suggest contacting your Thermo Electron agent or email > them on clinical.diagnostics@thermo.com. > > Hope this helps > > Darren James > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Paula > Lucas > Sent: Thursday, 7 October 2004 2:49 a.m. > To: Histonet (E-mail) > Subject: [Histonet] Cassette Label System > > > Hello, > > My company is possibly interested in purchasing a histology cassette > printer. > > Could you please provide information on the different manufactures that > offer printers? What are the pro/cons? Is it really that worth it to > purchase one? Does it really save that much time? Are they easy to use? > > Any information, suggestions, and comments are greatly appreciated. > > Thank you, > > Paula Lucas > Bio-Path Medical Group > > > _______________________________________________ > 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 > > -- Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax From ploykasek <@t> phenopath.com Wed Oct 6 18:01:01 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] antibody organizers In-Reply-To: <1453BDD1-17D2-11D9-8A9D-0005029D8A73@cvm.tamu.edu> Message-ID: We buy scintillation tubes that come in a great box with little dividers so that different size tubes can fit. I've also used the tackle or needlework craft organizers. Are all your tubes the same size or different sizes? Patti Loykasek Phenopath Laboratories Seattle, WA > Does anyone know of anyone that sells organizers/storage for stock > antibody vials? If anyone has a creative solution that would be great > also. Thanks > > Susan Hernandez > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JQB7 <@t> CDC.GOV Wed Oct 6 18:23:09 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Cassette Label System Message-ID: Sorry! I was talking about the slide laberler, not the cassette labeler. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bartlett, Jeanine Sent: Wed 10/6/2004 6:23 PM To: darrenj@medica.co.nz; Paula Lucas; Histonet (E-mail) Cc: Subject: RE: [Histonet] Cassette Label System I saw the ThermoElectron unit demo'd at NSH a few weeks ago. It looked really amazing. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Darren James Sent: Wed 10/6/2004 3:50 PM To: 'Paula Lucas'; Histonet (E-mail) Cc: Subject: RE: [Histonet] Cassette Label System Hi Paula, As agents for Thermo Electron in New Zealand I am going to extoll the virutes of the Shandon models. Unlike some on the market the Shandon uses foil tape and heated stylus. This burns the info into the cassette. They can print conventional and 2D barcodes and manage to print up to 3 lines of 19 characters. The software is fully configurable to customise the printing to your requirements. I have contacted several users througout NZ and Australia who all love the product. A couple of labs have 2 or 3 running at the same time. I think the biggest benefit is having totally legible cassettes and therefore a clear audit trail. Other users may like to comment. To get more info I suggest contacting your Thermo Electron agent or email them on clinical.diagnostics@thermo.com. Hope this helps Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Paula Lucas Sent: Thursday, 7 October 2004 2:49 a.m. To: Histonet (E-mail) Subject: [Histonet] Cassette Label System Hello, My company is possibly interested in purchasing a histology cassette printer. Could you please provide information on the different manufactures that offer printers? What are the pro/cons? Is it really that worth it to purchase one? Does it really save that much time? Are they easy to use? Any information, suggestions, and comments are greatly appreciated. Thank you, Paula Lucas Bio-Path Medical Group _______________________________________________ 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 From darrenj <@t> medica.co.nz Wed Oct 6 18:34:59 2004 From: darrenj <@t> medica.co.nz (Darren James) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Cassette Label System In-Reply-To: Message-ID: <002801c4abfd$18ea4b60$c864a8c0@medica.co.nz> RE: [Histonet] Cassette Label SystemThermo Electron (Shandon) also have a slide labeller. I have not seen this in action but it looks good on paper. -----Original Message----- From: Bartlett, Jeanine [mailto:JQB7@CDC.GOV] Sent: Thursday, 7 October 2004 12:23 p.m. To: Bartlett, Jeanine; darrenj@medica.co.nz; Paula Lucas; Histonet (E-mail) Subject: RE: [Histonet] Cassette Label System Sorry! I was talking about the slide laberler, not the cassette labeler. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bartlett, Jeanine Sent: Wed 10/6/2004 6:23 PM To: darrenj@medica.co.nz; Paula Lucas; Histonet (E-mail) Cc: Subject: RE: [Histonet] Cassette Label System I saw the ThermoElectron unit demo'd at NSH a few weeks ago. It looked really amazing. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Darren James Sent: Wed 10/6/2004 3:50 PM To: 'Paula Lucas'; Histonet (E-mail) Cc: Subject: RE: [Histonet] Cassette Label System Hi Paula, As agents for Thermo Electron in New Zealand I am going to extoll the virutes of the Shandon models. Unlike some on the market the Shandon uses foil tape and heated stylus. This burns the info into the cassette. They can print conventional and 2D barcodes and manage to print up to 3 lines of 19 characters. The software is fully configurable to customise the printing to your requirements. I have contacted several users througout NZ and Australia who all love the product. A couple of labs have 2 or 3 running at the same time. I think the biggest benefit is having totally legible cassettes and therefore a clear audit trail. Other users may like to comment. To get more info I suggest contacting your Thermo Electron agent or email them on clinical.diagnostics@thermo.com. Hope this helps Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Paula Lucas Sent: Thursday, 7 October 2004 2:49 a.m. To: Histonet (E-mail) Subject: [Histonet] Cassette Label System Hello, My company is possibly interested in purchasing a histology cassette printer. Could you please provide information on the different manufactures that offer printers? What are the pro/cons? Is it really that worth it to purchase one? Does it really save that much time? Are they easy to use? Any information, suggestions, and comments are greatly appreciated. Thank you, Paula Lucas Bio-Path Medical Group _______________________________________________ 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 From debbiekeith <@t> cox.net Wed Oct 6 19:21:13 2004 From: debbiekeith <@t> cox.net (Debbie Keith) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Mohs Tech needed in Peoria, AZ Message-ID: <5.2.0.9.0.20041006171310.020ac090@pop.central.cox.net> HI! I work for a busy (established) Mohs Surgeon in Peoria, Arizona. I am leaving on maternity leave shortly.. and will not be coming back. We are looking for a skilled tech that can make this transition easy for the Surgeon and the staff. Job title: MOHS Histotechnologist Description of job and areas of expertise required or preferred: Proper preparation of slides during MOHS Surgery following protocol to embed, cut and stain tissue, Maintain quality control logs and maintenance of cryostat. Certified Histotech preferred. Must be a self-directed individual with good organizational skills. Days: Mon-Wed 7am to 5pm. General: Excellent opportunity with well established, fellowship-trained practitioner. Position available immediately. Debbie please e-mail me at debbiekeith@cox.net if you have any questions OR fax your resume to Julie at 623-875-2621. thank you! :) From histophilhuff <@t> yahoo.com Thu Oct 7 01:29:46 2004 From: histophilhuff <@t> yahoo.com (Phillip Huff) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] antibody organizers - database for FileMaker Pro7 In-Reply-To: <1453BDD1-17D2-11D9-8A9D-0005029D8A73@cvm.tamu.edu> Message-ID: <20041007062946.723.qmail@web50302.mail.yahoo.com> Our lab uses rectangular plastic containers with lids that have a cardboard grid-like insert inside with each hole in the grid designated by a letter/number designation, ie. top left is A1 bottom right is E12. We then have numbered each of these boxes, designating each box for a specific type of antibody (ie. box 1 is alkaline phosphatase conjugated Abs) and have boxes in the 4C fridge, -20 freezer and -80 freezer. I then developed a database file for FileMaker Pro7 that organizes all of the antibodies so they are searchable by name/antigen, animal origin, conjugation, date purchased, company or any other characteristic you want. There are fields for catalog number, price, company and numerous other characteristics that makes searching for your antibody easy. You can even add notes about recommended dilution or pictures of optimal section staining. There is also an option to print a complete list in tabular form of all the antibodies so you can have a copy at the bench. If anyone is interested I can email a Mac or PC friendly version of the file, however you will need FileMaker Pro 7 for either respective OS. You can download FileMaker Pro 7 from the company's website and use it for 1 month for free just to see if the database for antibodies is practical for your usage. Phil I will look for the catalog number and company that sells the boxes tommorrow and post it. Histopath wrote: Does anyone know of anyone that sells organizers/storage for stock antibody vials? If anyone has a creative solution that would be great also. Thanks Susan Hernandez _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From lpwenk <@t> sbcglobal.net Thu Oct 7 04:18:27 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Results References: Message-ID: <007401c4ac4e$9c84ec40$78e0d445@domainnotset.invalid> Earliest for getting HT/HTL exam results - my guess - mid-January. The slides were due in Oct. 1. They will be graded the end of October. It normally takes around an additional 6-8 weeks after grading for ASCP to validate the grading. However, in the last 2 cycles, it has been taking up to an additional 4 weeks (10-12 weeks total after the grading date) before my students received their results. This is due to the extremely high number of people taking the exams, trying to get in before the Jan. 1, 2005 deadline where the high school route will be dropped. So, add 10 weeks onto the end of October grading, makes it about mid-January. At the earliest. Peggy A. Wenk, HTL(ASCP)SLS Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Fran Lemons" To: Sent: Wednesday, October 06, 2004 3:00 PM Subject: [Histonet] Results Hello fellow netters...... Can anyone give me an educated guess as to when we can expect results for the practical exam for this cycle? I have 2 techs on pins & needles waiting to hear. They already have results for the written, but we were wondering how long before the practical gets graded & we hear anything. Thanks Fran Walker HTS Baptist Hospital of E. TN (Knoxville) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MAUGER <@t> email.chop.edu Thu Oct 7 06:43:38 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] 4% PFA Message-ID: 4% PFA may be a good fixative in research setinng, but if the tissue is routinely processed through alcohols to xylene, and embedded in paraffin, the mouse brain tissue will be 'mushy' and impossible to cut well. If the same tissue is re-fixed in 10% NBF and processed,the cut sections will be great! Jo >>> "Colleen Forster" 10/06/04 05:26PM >>> 4% PFA is a very good fixative. The difference between 4% and 10% is that the PFA is made fresh each time and has no impurities added ex: methanols. The formalin content in both comes out to about the same. It is a standard fixative for many research labs. You can drop fix with PFA or perfuse and then drop fix for additional time for very good fixation and excellent results. I have used it for my mouse brain tissue for over 6 years with very nice results. Colleen Forster U of Mn _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MAUGER <@t> email.chop.edu Thu Oct 7 06:47:36 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] GI bx Message-ID: Michele, Could be crush artifact from clinicians or the gross bench. We use a shorter processing program for GI bxs. Make sure your 95% alc doesn't have methanol in it. This makes bxs very crispy(dries out too much)Drove us crazy too. Jo >>> "Michele Becker" 10/06/04 04:21PM >>> Please help.................... We are having problems with blurring and distorted nuclei on some of our GI biopsies. This has been happening on and off for the past few months. It is not every day and not all the GI bx on any given day. We have ruled out staining and processing. I really think it is something happening to the bxs before we even receive them but want to make sure we have not left any stones unturned on our end first. Any suggestions? Thank-you. Michele _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Thu Oct 7 06:50:28 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] GI bx Message-ID: I'll bet 5 bucks it goes back to your fixative - what type are you using? We experienced this problem when using zinc based fixatives with GI biopsies, and reverted back to plain old 10% NBF. Jackie O' "Michele Becker" Sent by: histonet-bounces@lists.utsouthwestern.edu 10/06/2004 03:21 PM To: "Histonet" cc: Subject: [Histonet] GI bx Please help.................... We are having problems with blurring and distorted nuclei on some of our GI biopsies. This has been happening on and off for the past few months. It is not every day and not all the GI bx on any given day. We have ruled out staining and processing. I really think it is something happening to the bxs before we even receive them but want to make sure we have not left any stones unturned on our end first. Any suggestions? Thank-you. Michele _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Thu Oct 7 07:07:35 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] GI bx Message-ID: <83AACDB0810528418AA106F9AE9B7F7E278451@sjhaexc02.sjha.org> This is the very first time I recall hearing that methanol might make them crispy! What kind of alcohols do you use? Thanks, j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joanne Mauger Sent: Thursday, October 07, 2004 7:48 AM To: mbecker@pathlabinc.com; histonet@pathology.swmed.edu Subject: Re: [Histonet] GI bx Michele, Could be crush artifact from clinicians or the gross bench. We use a shorter processing program for GI bxs. Make sure your 95% alc doesn't have methanol in it. This makes bxs very crispy(dries out too much)Drove us crazy too. Jo >>> "Michele Becker" 10/06/04 04:21PM >>> Please help.................... We are having problems with blurring and distorted nuclei on some of our GI biopsies. This has been happening on and off for the past few months. It is not every day and not all the GI bx on any given day. We have ruled out staining and processing. I really think it is something happening to the bxs before we even receive them but want to make sure we have not left any stones unturned on our end first. Any suggestions? Thank-you. Michele _______________________________________________ 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 Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From tsalavos <@t> fleming.gr Thu Oct 7 07:09:13 2004 From: tsalavos <@t> fleming.gr (Sotiris Tsalavos) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Background with DAB staining Message-ID: <8AE623DD13E596459F855D6C67F9850A0A2F51@server.fleming.local> Hallo. I'm currently trying out to stain murine spleen and thymous for dendritic cells using the hamster anti-mouseCD11c antibody. As a secondary Ab I use biotin goat anti-hamster IgG and then streptavidin HRP for DAB staining. I'm getting a lot of background in both types of tissues even in the negative control. Below I have the protocol that I use. Frozen tissues cut at 10um placed on gelatin embedded slides. Air dry for 30min, fix in cold acetone for 10min and then in PBS/0.3% H2O2 for 10min for the endogenous peroxidase. Slides blocked with PBS/5% Goat serum for 30min. Primary Ab anti-mouse CD11c at 1:100 dilution for 1 hour. Secondary anti-hamster IgG at 1:800 dilution for 1 hour. Streptavidin HRP at 1:500 for 30min. Use DAB prepared solution 0.05% DAB- 0.015% H2O2 in 0.01M PBS pH 6.8 The DAB I use is from Sigma. I'm not sure that the background is still from the endogenous peroxidases in the tissues or something else. I have tried staining the tissues with just the secondary Ab from 1:200 up to 1:3200 dilution. In the high 1:3200 get very little background however I'm afraid that it might be too dilute to bind the primary Ab. Any suggestions would be greatly appreciated. Thanks Sotiris Tsalavos Institute of Immunology B.S.R.C. Alexander Fleming Athens, Greece tsalavos@fleming,gr From sladd <@t> hsc.usf.edu Thu Oct 7 07:17:24 2004 From: sladd <@t> hsc.usf.edu (S Ladd) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] 4% PFA In-Reply-To: Message-ID: I completely agree with you. I have done side by side experiments and found the results to be far superior with 10% NBF or 10% formalin. For whatever reason, the sections expand and distort more on the waterbath with 4% formaldehyde made from paraformaldehyde (4%PFA) and are more difficult to section. I think its the methanol (the very methanol everyone is trying to avoid by using 4%PFA). Sharron USF -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joanne Mauger Sent: Thursday, October 07, 2004 7:44 AM To: histonet@pathology.swmed.edu; cforster@umn.edu Subject: Re: [Histonet] 4% PFA 4% PFA may be a good fixative in research setinng, but if the tissue is routinely processed through alcohols to xylene, and embedded in paraffin, the mouse brain tissue will be 'mushy' and impossible to cut well. If the same tissue is re-fixed in 10% NBF and processed,the cut sections will be great! Jo >>> "Colleen Forster" 10/06/04 05:26PM >>> 4% PFA is a very good fixative. The difference between 4% and 10% is that the PFA is made fresh each time and has no impurities added ex: methanols. The formalin content in both comes out to about the same. It is a standard fixative for many research labs. You can drop fix with PFA or perfuse and then drop fix for additional time for very good fixation and excellent results. I have used it for my mouse brain tissue for over 6 years with very nice results. Colleen Forster U of Mn _______________________________________________ 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 From Jackie.O'Connor <@t> abbott.com Thu Oct 7 07:20:54 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Background with DAB staining Message-ID: You're using an SABC protocol on biotin-rich (at least the spleen) tissues - you're probably seeing endogenous biotin background. Use an avidin/biotin block (commercially available from several suppliers, Biocare is one, Vector another). Jackie O'Connor Abbott Laboratories Global Pharmaceutical Research and Development "Sotiris Tsalavos" Sent by: histonet-bounces@lists.utsouthwestern.edu 10/07/2004 07:09 AM To: cc: Subject: [Histonet] Background with DAB staining Hallo. I'm currently trying out to stain murine spleen and thymous for dendritic cells using the hamster anti-mouseCD11c antibody. As a secondary Ab I use biotin goat anti-hamster IgG and then streptavidin HRP for DAB staining. I'm getting a lot of background in both types of tissues even in the negative control. Below I have the protocol that I use. Frozen tissues cut at 10um placed on gelatin embedded slides. Air dry for 30min, fix in cold acetone for 10min and then in PBS/0.3% H2O2 for 10min for the endogenous peroxidase. Slides blocked with PBS/5% Goat serum for 30min. Primary Ab anti-mouse CD11c at 1:100 dilution for 1 hour. Secondary anti-hamster IgG at 1:800 dilution for 1 hour. Streptavidin HRP at 1:500 for 30min. Use DAB prepared solution 0.05% DAB- 0.015% H2O2 in 0.01M PBS pH 6.8 The DAB I use is from Sigma. I'm not sure that the background is still from the endogenous peroxidases in the tissues or something else. I have tried staining the tissues with just the secondary Ab from 1:200 up to 1:3200 dilution. In the high 1:3200 get very little background however I'm afraid that it might be too dilute to bind the primary Ab. Any suggestions would be greatly appreciated. Thanks Sotiris Tsalavos Institute of Immunology B.S.R.C. Alexander Fleming Athens, Greece tsalavos@fleming,gr _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Mandy <@t> serotec.co.uk Thu Oct 7 07:25:03 2004 From: Mandy <@t> serotec.co.uk (Mandy Townsend) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Background with DAB staining Message-ID: Sounds like your secondary is reacting with your tissue, unless it has been claimed that is has been adsorbed against mouse. therefore you should try diluting the secondary in mouse serum to remove any cross reactivity, then hopefully your sections with just secondary will come out negative. mandy -----Original Message----- From: Sotiris Tsalavos [mailto:tsalavos@fleming.gr] Sent: Thursday, October 07, 2004 1:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Background with DAB staining Hallo. I'm currently trying out to stain murine spleen and thymous for dendritic cells using the hamster anti-mouseCD11c antibody. As a secondary Ab I use biotin goat anti-hamster IgG and then streptavidin HRP for DAB staining. I'm getting a lot of background in both types of tissues even in the negative control. Below I have the protocol that I use. Frozen tissues cut at 10um placed on gelatin embedded slides. Air dry for 30min, fix in cold acetone for 10min and then in PBS/0.3% H2O2 for 10min for the endogenous peroxidase. Slides blocked with PBS/5% Goat serum for 30min. Primary Ab anti-mouse CD11c at 1:100 dilution for 1 hour. Secondary anti-hamster IgG at 1:800 dilution for 1 hour. Streptavidin HRP at 1:500 for 30min. Use DAB prepared solution 0.05% DAB- 0.015% H2O2 in 0.01M PBS pH 6.8 The DAB I use is from Sigma. I'm not sure that the background is still from the endogenous peroxidases in the tissues or something else. I have tried staining the tissues with just the secondary Ab from 1:200 up to 1:3200 dilution. In the high 1:3200 get very little background however I'm afraid that it might be too dilute to bind the primary Ab. Any suggestions would be greatly appreciated. Thanks Sotiris Tsalavos Institute of Immunology B.S.R.C. Alexander Fleming Athens, Greece tsalavos@fleming,gr _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________ This e-mail has been scanned for all viruses by Star. The service is powered by MessageLabs. For more information on a proactive anti-virus service working around the clock, around the globe, visit: http://www.star.net.uk ________________________________________________________________________ From MAUGER <@t> email.chop.edu Thu Oct 7 07:28:07 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] GI bx Message-ID: We use 95% ethanol without methanol added, but our absolute does contain a small amount. For us, changing the 95% made all the difference. we had switched companies, and didn't realize that most ethanols had ethanol in them. Our bxs got real crunchy. When we read the 95 ingredients more carefully, we found the culprit, and when we replaced it with the non meth. 95 the bxs were gorgeous again! Our pharmacy orders the 95% from Aaper in Shelbyville,Kentucky. jo >>> "Weems, Joyce" 10/07/04 08:07AM >>> This is the very first time I recall hearing that methanol might make them crispy! What kind of alcohols do you use? Thanks, j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joanne Mauger Sent: Thursday, October 07, 2004 7:48 AM To: mbecker@pathlabinc.com; histonet@pathology.swmed.edu Subject: Re: [Histonet] GI bx Michele, Could be crush artifact from clinicians or the gross bench. We use a shorter processing program for GI bxs. Make sure your 95% alc doesn't have methanol in it. This makes bxs very crispy(dries out too much)Drove us crazy too. Jo >>> "Michele Becker" 10/06/04 04:21PM >>> Please help.................... We are having problems with blurring and distorted nuclei on some of our GI biopsies. This has been happening on and off for the past few months. It is not every day and not all the GI bx on any given day. We have ruled out staining and processing. I really think it is something happening to the bxs before we even receive them but want to make sure we have not left any stones unturned on our end first. Any suggestions? Thank-you. Michele _______________________________________________ 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 Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From pmarcum <@t> polysciences.com Thu Oct 7 08:00:16 2004 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] GI bx In-Reply-To: Message-ID: <001e01c4ac6d$97792c10$4f00a8c0@PMARCUM2K> Actually you may need to check that even more fully. The SDA 3 alcohols we use as reagent alcohol to avoid tax and the ATF contains both isopropanol and methanol at 3% to 5% of each type depending on the supplier and the license they have for manufacture. In a university or non-taxable institution you can afford the really good pure stuff while the tax often means the rest of us have to find something cheaper and carefully revamp our processing and thinking. I know there are other netters who can explain this in far more detail however this is the basic. Thanks, Pam Marcum? > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joanne > Mauger > Sent: Thursday, October 07, 2004 8:28 AM > To: mbecker@pathlabinc.com; histonet@pathology.swmed.edu; > JWEEMS@sjha.org > Subject: RE: [Histonet] GI bx > > > We use 95% ethanol without methanol added, but our absolute does > contain a small amount. For us, changing the 95% made all the > difference. we had switched companies, and didn't realize that > most ethanols had ethanol in them. Our bxs got real crunchy. When > we read the 95 ingredients more carefully, we found the culprit, > and when we replaced it with the non meth. 95 the bxs were > gorgeous again! Our pharmacy orders the 95% from Aaper in > Shelbyville,Kentucky. > jo > > >>> "Weems, Joyce" 10/07/04 08:07AM >>> > This is the very first time I recall hearing that methanol might > make them crispy! What kind of alcohols do you use? > > Thanks, j > > Joyce Weems > Pathology Manager > Saint Joseph's Hospital > 5665 Peachtree Dunwoody Rd NE > Atlanta, GA 30342 > 404-851-7376 - Phone > 404-851-7831 - Fax > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joanne > Mauger > Sent: Thursday, October 07, 2004 7:48 AM > To: mbecker@pathlabinc.com; histonet@pathology.swmed.edu > Subject: Re: [Histonet] GI bx > > > Michele, > Could be crush artifact from clinicians or the gross bench. We > use a shorter processing program for GI bxs. Make sure your 95% > alc doesn't have methanol in it. This makes bxs very crispy(dries > out too much)Drove us crazy too. > Jo > > >>> "Michele Becker" 10/06/04 04:21PM >>> > Please help.................... We are having problems with blurring and > distorted nuclei on some of our GI biopsies. This has been > happening on and > off for the past few months. It is not every day and not all the GI bx on > any given day. We have ruled out staining and processing. I > really think it > is something happening to the bxs before we even receive them but want to > make sure we have not left any stones unturned on our end first. Any > suggestions? > > Thank-you. > > Michele > > > > _______________________________________________ > 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 > > > Confidentiality Notice ** The information contained in this > message may be privileged and is confidential information > intended for the use of the addressee listed above. If you are > neither the intended recipient nor the employee or agent > responsible for delivering this message to the intended > recipient, you are hereby notified that any disclosure, copying, > distribution or the taking of any action in reliance on the > contents of this information is strictly prohibited. If you have > received this communication in error, please notify us > immediately by replying to the message and deleting it from your computer. > Thank you. Saint Josephs Health System, Inc. > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From dobbin <@t> upei.ca Thu Oct 7 08:01:59 2004 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Background with DAB staining In-Reply-To: Message-ID: <41651416.19366.3824CB@localhost> Sotiris, Have you considered blocking for endogenous biotin? Greg Date sent: Thu, 7 Oct 2004 13:25:03 +0100 From: "Mandy Townsend" To: "Sotiris Tsalavos" , Copies to: Subject: RE: [Histonet] Background with DAB staining > Sounds like your secondary is reacting with your tissue, unless it has been claimed that is has been adsorbed against mouse. therefore you should try diluting the secondary in mouse serum to remove any cross reactivity, then hopefully your sections with just secondary will come out negative. > > mandy > > -----Original Message----- > From: Sotiris Tsalavos [mailto:tsalavos@fleming.gr] > Sent: Thursday, October 07, 2004 1:09 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Background with DAB staining > > > Hallo. > > I'm currently trying out to stain murine spleen and thymous for > dendritic cells using the hamster anti-mouseCD11c antibody. > > As a secondary Ab I use biotin goat anti-hamster IgG and then > streptavidin HRP for DAB staining. I'm getting a lot of background in > both types of tissues even in the negative control. Below I have the > protocol that I use. > > > > Frozen tissues cut at 10um placed on gelatin embedded slides. > > Air dry for 30min, fix in cold acetone for 10min and then in PBS/0.3% > H2O2 for 10min for the endogenous peroxidase. > > Slides blocked with PBS/5% Goat serum for 30min. > > Primary Ab anti-mouse CD11c at 1:100 dilution for 1 hour. > > Secondary anti-hamster IgG at 1:800 dilution for 1 hour. > > Streptavidin HRP at 1:500 for 30min. > > Use DAB prepared solution 0.05% DAB- 0.015% H2O2 in 0.01M PBS pH 6.8 > > > > The DAB I use is from Sigma. > > I'm not sure that the background is still from the endogenous > peroxidases in the tissues or something else. > > I have tried staining the tissues with just the secondary Ab from 1:200 > up to 1:3200 dilution. In the high 1:3200 get very little background > however I'm afraid that it might be too dilute to bind the primary Ab. > > > > Any suggestions would be greatly appreciated. Thanks > > > > Sotiris Tsalavos > > Institute of Immunology > > B.S.R.C. Alexander Fleming > > Athens, Greece > > > > tsalavos@fleming,gr > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ________________________________________________________________________ > This e-mail has been scanned for all viruses by Star. The > service is powered by MessageLabs. For more information on a proactive > anti-virus service working around the clock, around the globe, visit: > http://www.star.net.uk > ________________________________________________________________________ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Pathology Lab Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Happiness is a journey, not a destination. From psanquin <@t> lugo.usc.es Thu Oct 7 09:52:44 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] 4% PFA (Carcinogenic?) In-Reply-To: References: Message-ID: <3.0.6.32.20041007165244.00805920@pop.lugo.usc.es> Whenever I have had problems with 4% PFA samples (mainly in brain samples) I have turned to Bouin's Fluid. It is wonderful: much easier to prepare than paraformaldehyde and fixation is really good with excellent preservation of the tissue and even nicer H-E staining than PFA. On the negative side some histological stainings and some antibodies do not work with Bouin's samples. But if you have problems with PFA is worth to have a trial with Bouin. On the negative side of PFA I have recently read that PFA is probably carcinogenic in humnas. Is that true? What cautions do you take, PFA people? During perfusion and dissection of the tissue it is not easy to me avoiding contact with the PFA fumes. Thanks for your input. Pablo Sanchez >I completely agree with you. I have done side by side experiments and found >the results to be far superior with 10% NBF or 10% formalin. For whatever >reason, the sections expand and distort more on the waterbath with 4% >formaldehyde made from paraformaldehyde (4%PFA) and are more difficult to >section. I think its the methanol (the very methanol everyone is trying to >avoid by using 4%PFA). > >Sharron >USF From pmarcum <@t> polysciences.com Thu Oct 7 09:46:03 2004 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] 4% PFA (Carcinogenic?) In-Reply-To: <3.0.6.32.20041007165244.00805920@pop.lugo.usc.es> Message-ID: <002a01c4ac7c$5e7a44d0$4f00a8c0@PMARCUM2K> Paraformaldehyde is the powder form of Formaldehyde so the rules for both are to be carefully considered and compared. We have many things we work with daily in Histology or any laboratory that are known or suspected carcinogens and should be handled as such. Always read your MDSD paperwork with new products or applications for warnings. It is wise to periodically look at standard products for as we sometimes become complacent and forget the dangers. Best Regards, Pamela A Marcum Histology/Microscopy Product Development Manager Polysciences, Inc. 400 Valley Road Warrington, PA 18976 Telephone: 800-523-2575???? Ext. 167 215-343-6484???? Ext. 167 Fax:? 800-343-3291 ???????? 215-343-0214 > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Pablo > S?nchez Quinteiro > Sent: Thursday, October 07, 2004 10:53 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] 4% PFA (Carcinogenic?) > > > Whenever I have had problems with 4% PFA samples (mainly in brain samples) > I have turned to Bouin's Fluid. It is wonderful: much easier to prepare > than paraformaldehyde and fixation is really good with excellent > preservation of the tissue and even nicer H-E staining than PFA. On the > negative side some histological stainings and some antibodies do not work > with Bouin's samples. But if you have problems with PFA is worth to have a > trial with Bouin. > > On the negative side of PFA I have recently read that PFA is probably > carcinogenic in humnas. Is that true? What cautions do you take, PFA > people? During perfusion and dissection of the tissue it is not easy to me > avoiding contact with the PFA fumes. > > Thanks for your input. > > Pablo Sanchez > > > >I completely agree with you. I have done side by side > experiments and found > >the results to be far superior with 10% NBF or 10% formalin. For whatever > >reason, the sections expand and distort more on the waterbath with 4% > >formaldehyde made from paraformaldehyde (4%PFA) and are more difficult to > >section. I think its the methanol (the very methanol everyone is > trying to > >avoid by using 4%PFA). > > > >Sharron > >USF > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From ajennings <@t> unmc.edu Thu Oct 7 09:54:13 2004 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Creative people needed In-Reply-To: <007401c4ac4e$9c84ec40$78e0d445@domainnotset.invalid> Message-ID: Most of you know that we are in the process of planning a hub where histonetters can gather at the 2005 C/S in Florida. Our "OutPost" for mingling. One of the suggestions that I am following up on is the distribution of 'Histonet' buttons. I would like help on the design. Send them to me personally, NOT via the histonet listserv, please. Preferably as .jpeg. Not real sure if you will receive anything except recognition from your fellow histonetters and my undying gratitude, that will be decided after we have a sponsor for the production costs. If possible the "winning" design should at least receive a few of the buttons for their own distribution. The only requirements for the buttons that I can see (open to suggestions) would be >something that catches your eye as unique so as to identify the wearer as a histonetter >the url for Histonet http://www.histonet.org/ > a 'white space' for writing in your name or screen name with a sharpie/marker (does not have to be white) >name of the company that sponsors the production I will pull out my old buttons and scan them to give you an idea of what we had before, if you need to see them for inspiration. Right now I just need your input on design Things we will discuss after the first of the year..... >'winning design' >people who can not make it to the C/S but would still like a button >number produced and distribution requirements Please save your comments/suggestions on these topics until I post the discussion after the first of the year. We have two companies represented in our work group, but I don't want to obligate them to "volunteer" their companies, since they are already helping so much on the team. If a company wants to be considered for production expenses they can contact me personally. From sladd <@t> hsc.usf.edu Thu Oct 7 10:03:37 2004 From: sladd <@t> hsc.usf.edu (S Ladd) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] 4% PFA (Carcinogenic?) In-Reply-To: <3.0.6.32.20041007165244.00805920@pop.lugo.usc.es> Message-ID: We do perfusions in a fume hood. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Pablo Sanchez Quinteiro Sent: Thursday, October 07, 2004 10:53 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] 4% PFA (Carcinogenic?) Whenever I have had problems with 4% PFA samples (mainly in brain samples) I have turned to Bouin's Fluid. It is wonderful: much easier to prepare than paraformaldehyde and fixation is really good with excellent preservation of the tissue and even nicer H-E staining than PFA. On the negative side some histological stainings and some antibodies do not work with Bouin's samples. But if you have problems with PFA is worth to have a trial with Bouin. On the negative side of PFA I have recently read that PFA is probably carcinogenic in humnas. Is that true? What cautions do you take, PFA people? During perfusion and dissection of the tissue it is not easy to me avoiding contact with the PFA fumes. Thanks for your input. Pablo Sanchez >I completely agree with you. I have done side by side experiments and found >the results to be far superior with 10% NBF or 10% formalin. For whatever >reason, the sections expand and distort more on the waterbath with 4% >formaldehyde made from paraformaldehyde (4%PFA) and are more difficult to >section. I think its the methanol (the very methanol everyone is trying to >avoid by using 4%PFA). > >Sharron >USF _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eenriquez52234 <@t> yahoo.com Thu Oct 7 10:06:12 2004 From: eenriquez52234 <@t> yahoo.com (esteban enriquez) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] tissue processors In-Reply-To: Message-ID: <20041007150612.52609.qmail@web61110.mail.yahoo.com> Tissue Tek VIPs is reliable, they don't has the problems with line clogging continually like some Company's Polaris tissue processor. What the good of higher throughput if the thing is always needing to be fixed??? I stay with the Tissue Tek. EE "GUTIERREZ, JUAN" wrote: Nothing beats the Tissue Teks. They keep going and going and going. Sorry but I can not say anything bad about equipment or companies anymore. Too many damn lawyers here in the USofA. Juan -----Original Message----- From: Andres Kulla [mailto:Andres.Kulla@kliinikum.ee] Sent: Tuesday, October 05, 2004 1:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] tissue processors To the experts in automated tissue processing, Could anybody share his/her experience with TBS (Triangle Biomedical Sciences) ATP1 tissue processor. Positive vs negative aspects in comparison with other devices eg Sakuras Tissue Tek VIP 500. With best regards from Andres Kulla, Estonia _______________________________________________ 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 --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From gcallis <@t> montana.edu Thu Oct 7 10:11:41 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Background with DAB staining In-Reply-To: <8AE623DD13E596459F855D6C67F9850A0A2F51@server.fleming.loca l> References: <8AE623DD13E596459F855D6C67F9850A0A2F51@server.fleming.local> Message-ID: <6.0.0.22.1.20041007083506.01b45110@gemini.msu.montana.edu> We do CD11c antibody all the time on frozen sections, and do not get background. First, cut your sections at 5 um instead of 10 um, 6 um is ok too. Pick up your sections on Silane coated or plus charged slides or poly L lysine coated but NOT gelatin coated. You can prepare these slides yourself, HISTONET has methods to use, do a search in Histonet Archives www.histosearch.org. Air dry sections overnight, then fix in an acetone alcohol mixture at RT for 5 min then go directly to 3 changes buffer. Do NOT air dry again. Proceed with staining. Our staining buffers whether we use either TRIS buffered saline OR Dulbeccos PBS (Sigma) contain 0.2% goat serum and 0.05% Tween 20. All diluents, normal serum blocks are made up in this buffer. We prefer the Glucose oxidase peroxidase block for cleaning up endogenous peroxidase particularly for dendritic cell staining, if you want - I will attach to you privately. Most of the time we have success with DAKO peroxidase blocker, S2001 for 10 - 15 minutes, we do not follow their instructions for 5 minutes, and it has NOT hurt our staining but does a more efficienct job of quenching endog peroxidase. On thing to remember, CD11c is a clone from Armenian hamster and you must use a secondary that detects the Armenian hamster, and the secondary MUST be adsorbed to the mouse. There are two sources of secondaries we use, our favorite is Jackson Immunoresearch, Goat antiArmenian hamster, adsorbed to mouse Cat #127-065-160 and BD Pharmingen has a cocktail to detect Armenian AND Golden Syrian Cat #550335and this one will work nicely too. Both are biotinylated and used diluted 1:250 or so. If your antibody is NOT adsorbed against mouse, this may be a major source of your background. If you already have an antiArmenian hamster secondary, you can do the adsorption yourself by diluting your secondary antibody in 1 or 2% mouse serum, let it stand for an hour or so and then apply it to the mouse spleen. We prefer to buy the antibody already adsorbed and stay away from mouse serum with hamster antibodies, less work, reliable. Our staining protocol: Fix, block endogenous peroxidase (yours should work as long as you don't chew section off with higher conc of hydrogen peroxide). I discussed this previously. Normal serum block - yours looks fine, we go as high at 10% goat serum but we heat inactivate our normal serums to get rid of complement. Do an Avidin/biotin block!! Vector has a kit. CD11c, your antibody concentration is in the ballpark, but we only incubate 30 min at RT. This antibody is diluted in 1% goat serum. Secondary, goat antiArmenian Hamster, adsorbed to mouse, Jackson or BD Pharm cocktain diluted 1:250 or so, 30 min,. This is diluted in the NORMAL SERUM BLOCK of 5% goat. Strepavidin-HRP (TAGO/Biosource) diluted 1:500 (0.5 to 0.6 mg/ml original concentration) 20 min Rinse with buffer containing only 0.05% Tween 20, and add chromogen. Develop your POSITIVE control using a microscope, no exceptions in this lab. We let a timer run to have approx time just for record keeping, an awareness of what to expect for future stainings. We prefer AEC to DAB, better contrast. We like Chris van der Loos's AEC, excellent!! Rinse off DAB, and proceed with hematoxylin counterstain, very light. If you think your DAB is giving you trouble, try others, DAKO DAB+ or Pierce DAB two part system or Pierce DAB enhanced for black ppt. If the chromogen develops too fast, you may have to readjust the primary concentration. Did you do a dilution panel to determine optimal working concentration of your primary? If not, you may have it too concentrated for your other reagents, particularly since you are using DAB. For dilution panel, work with 10 ug/ml and do dilutions from there. You may be surprised at results and also eliminate background. Good luck Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman Bozeman MT 59717-3610 At 06:09 AM 10/7/2004, you wrote: >Hallo. > >I'm currently trying out to stain murine spleen and thymous for >dendritic cells using the hamster anti-mouseCD11c antibody. > >As a secondary Ab I use biotin goat anti-hamster IgG and then >streptavidin HRP for DAB staining. I'm getting a lot of background in >both types of tissues even in the negative control. Below I have the >protocol that I use. > >Frozen tissues cut at 10um placed on gelatin embedded slides. > >Air dry for 30min, fix in cold acetone for 10min and then in PBS/0.3% >H2O2 for 10min for the endogenous peroxidase. > >Slides blocked with PBS/5% Goat serum for 30min. > >Primary Ab anti-mouse CD11c at 1:100 dilution for 1 hour. > >Secondary anti-hamster IgG at 1:800 dilution for 1 hour. > >Streptavidin HRP at 1:500 for 30min. > >Use DAB prepared solution 0.05% DAB- 0.015% H2O2 in 0.01M PBS pH 6.8 > > > >The DAB I use is from Sigma. > >I'm not sure that the background is still from the endogenous >peroxidases in the tissues or something else. > >I have tried staining the tissues with just the secondary Ab from 1:200 >up to 1:3200 dilution. In the high 1:3200 get very little background >however I'm afraid that it might be too dilute to bind the primary Ab. > > > >Any suggestions would be greatly appreciated. Thanks > > > >Sotiris Tsalavos > >Institute of Immunology > >B.S.R.C. Alexander Fleming > >Athens, Greece > > > >tsalavos@fleming,gr > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eenriquez52234 <@t> yahoo.com Thu Oct 7 10:10:47 2004 From: eenriquez52234 <@t> yahoo.com (esteban enriquez) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Ventana Wins! Message-ID: <20041007151047.54522.qmail@web61110.mail.yahoo.com> For those who hadnt check Ventana company site lately, they announced a preliminary victory over Vision Biosystem re bar code patent infringerment. No more Bond systems for sale in USA??????? EE --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From gentras <@t> vetmed.auburn.edu Thu Oct 7 10:26:33 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:24:08 2005 Subject: Fwd: Re: [Histonet] mouse brains Message-ID: <6.0.1.1.0.20041007102624.01b67188@mailhost.vetmed.auburn.edu> >Date: Thu, 07 Oct 2004 09:43:12 -0500 >To: meyenhmf@umdnj.edu >From: "Atoska S. Gentry" >Subject: Re: [Histonet] mouse brains > > > >Thanks for your inquiry. Acquiring fairly decent sections is involving a >lot more trial and error than previous sections processed & done a few >months back using the same procedure. Pointers & input will be much >appreciated. Atoska > >At 02:54 PM 10/6/2004, you wrote: >>What IS the problem?/ >> >>Quoting "Atoska S. Gentry" : >> >> > >> > Hello, those of you processing & sectioning coronal sections of mouse >> > brain; from you experience can prolonged fixation in 4%PFA be the >> > culprit >> > of sectioning problems? What is the maximum fixation time period the >> > brain >> > should held in PFA before processing & sectioning? Thanks! Atoska >> > >> > >> > Atoska S. Gentry B.S., HT(ASCP) >> > Research Assistant III >> > Scott-Ritchey Research Center >> > College of Veterinary Medicine >> > Auburn University, AL 36849 >> > Phone# (334)844-5579 Fax# (334)844-5850 >> > >> > >> > _______________________________________________ >> > Histonet mailing list >> > Histonet@lists.utsouthwestern.edu >> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > >> >> >>-- From gentras <@t> vetmed.auburn.edu Thu Oct 7 10:26:56 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:24:08 2005 Subject: Fwd: Re: [Histonet] mouse brains Message-ID: <6.0.1.1.0.20041007102646.01b59648@mailhost.vetmed.auburn.edu> >Date: Thu, 07 Oct 2004 10:26:10 -0500 >To: Gayle Callis >From: "Atoska S. Gentry" >Subject: Re: [Histonet] mouse brains > > >Gayle, thanks for your reply. Actually, I'm processing these samples for a >professor from the Biological Science Department on main campus. And she's >conducting various IHC stains for "Tau Filament Formation in Transgenic >Mice Expressing P301L Tau" & neurofibrillary changes in CNS. She's working >in conjunction with some Spanish Scientists. I believe the samples were >shipped from Spain in PFA back in February 04'. When we receive them they >are whole brains in PFA from which coronal slices are taken, processed, >and sectioned. The first few sets of brain received a few months ago >sectioned much easier, but the two she brought out a couple of weeks ago >are requiring a fair amount of trial & error to obtain fairly decent >sections. Atoska > > >At 03:11 PM 10/6/2004, you wrote: >>Atoska, >> >>More info please. Are you processing brain slices cut with a matrix >>device or whole brain. We do both and find the time in processing is >>critical. We just did whole mouse brains fixed for three weeks in NBF >>for LFB staining and H&E and had wonderful sections but used a longer >>processing schedule than for our coronal mouse brain slices. >> >>For our coronal sections, we perfuse after anesthetizing animal - first >>with saline followed by PLP via heart (a morning protocol then fix for an >>additonal 5 hours, slice into 3 mm thick slices with matrix device prior >>to a special mouse brain processing schedule on a VIP the same >>evening. PLP has paraformaldehyde so will be similar to your PFA >>fixation and are able to obtain excellent coronal sections. I have not >>noticed any differences between longer NBF nor PLP fixation on sectioning >>effects. >> >>If we fix brain with PFA, we do this overnight at 4C, and if the fixation >>needs to be longer, we change the PFA to fresh. Are you perfusing >>(ideal situation) then immersing samples overnight, immersion only, at RT >>or 4C. >> >>What sectioning problems do you encounter? >> >> >> >>At 01:08 PM 10/6/2004, you wrote: >> >>>Hello, those of you processing & sectioning coronal sections of mouse >>>brain; from you experience can prolonged fixation in 4%PFA be the >>>culprit of sectioning problems? What is the maximum fixation time period >>>the brain should held in PFA before processing & sectioning? Thanks! Atoska >>> >>> >>>Atoska S. Gentry B.S., HT(ASCP) >>>Research Assistant III >>>Scott-Ritchey Research Center >>>College of Veterinary Medicine >>>Auburn University, AL 36849 >>>Phone# (334)844-5579 Fax# (334)844-5850 >>> >>>_______________________________________________ >>>Histonet mailing list >>>Histonet@lists.utsouthwestern.edu >>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >>Gayle Callis >>MT,HT,HTL(ASCP) >>Research Histopathology Supervisor >>Veterinary Molecular Biology >>Montana State University - Bozeman >>PO Box 173610 >>Bozeman MT 59717-3610 >>406 994-6367 (lab with voice mail) >>406 994-4303 (FAX) From srishan <@t> mail.holyname.org Thu Oct 7 10:39:11 2004 From: srishan <@t> mail.holyname.org (srishan@mail.holyname.org) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Staining with recycled xylene and alcohol Message-ID: __________________ Hi All, currently we have a demo recycler for xylene and alcohol. The pathologist tend to see the H&E staining not as crisp as it used to be. Any suggestions and advice will be appreciated. Thanks in advance Nirmala Srishan Histology Supervisor Holy Name Hospital Teaneck NJ, 07666 From nperson211 <@t> comcast.net Thu Oct 7 12:08:56 2004 From: nperson211 <@t> comcast.net (nperson211@comcast.net) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] b-Gal antibody Message-ID: <100720041708.10111.41657828000A9ADD0000277F2200735446CECECD02019C9D0A9F02@comcast.net> Histonetters, Does anyone have a favorite b-gal antibody for use in FFPE tissues? I would really appreciate some information. Thanks in advance, Nancy Lemke Hermelin Brain Tumor Center Henry Ford Hospital Detroit From kalschev <@t> svm.vetmed.wisc.edu Thu Oct 7 13:17:10 2004 From: kalschev <@t> svm.vetmed.wisc.edu (Vicki Kalscheur) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Kermit Groothuis Memorial Scholarship Message-ID: <001301c4ac99$dcff9e50$65d56880@kalscheur426> Kermit Groothuis Honored at Retirement Reception Kermit Groothuis, an 18-year veteran at the University of Wisconsin-Madison School of Veterinary Medicine, retired from his position in histopathology in August. Even Bucky Badger, the University of Wisconsin mascot, came to share in the celebration. Over the years, Kermit made numerous friends at the SVM as well as state and national histology forums. In recognition, for his service, the Wisconsin Histotechnology Society and the National Society for Histotechnology, as well as colleagues from his labs purchased several SVM tiles in his name, honoring his many contributions. These tiles have been installed in front of the University of Wisconsin - School of Veterinary Medicine. A good time was had by all at the retirement reception. Regretfully, our friend and colleague, Kermit B. Groothuis, (HT. ASCP.), 64 years young, passed away after a short bout with cancer. A lover of all sports, he refereed team sports and worked UW football and basketball games, and was also active in the Boy Scouts. He enjoyed all people and was a friend to all. Our heartfelt condolences go out to his wife, Mary, and family members. Kermit B. Groothuis September 25, 1939-September 21, 2004 God Bless For interested parties, colleagues and friends: A Kermit Groothuis Memorial Scholarship is being established, for a veterinary student, to be awarded, each year at a graduation banquet. If you wish to contribute make checks payable to: University of Wisconsin Foundation. Mail to Att: Nancy J Nelson, School of Veterinary Medicine, 2015 Linden Dr. Madison, WI 53706. Or visit www.vetmed.wisc.edu to give online. Please specify the Kermit Groothuis Memorial Scholarship. Email questions to nelson@svm.vetmed.wisc.edu From RSRICHMOND <@t> aol.com Thu Oct 7 13:24:13 2004 From: RSRICHMOND <@t> aol.com (RSRICHMOND@aol.com) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Re: GI bx Message-ID: <06609E92.2D708D4B.0BF1BB09@aol.com> In regard to alcohols: Do people have really strong evidence that denatured alcohol behaves any differently from pure ethanol? Absolute (100%) ethanol is a major security problem for the laboratory, since tax authorities (Bureau of Alcohol, Tobacco, and Firearms - Department of Justice - in the USA) want to make really sure that it isn't diverted for beverage use. Many hospitals just don't have the storage facilities. In common use is what's generically called "reagent alcohol", with 90% ethanol, 5% methanol, and 5% isopropanol. This is denaturation formula "S3DA modified", the "modified" referring to the fact that isopropanol is also added to the methanol-denatured S3DA mix. - Another denaturant permitted in histology is methylisobutylketone (MIBK), which smells so bad that most labs don't want to use it. (Alcohol denatured with acetone is also available - unacceptable because it removes eosin instantly.) This old pathologist hasn't seen any difference between pure ethanol and reagent alcohol. Is any of you sure that there is a difference? Bob Richmond Samurai Pathologist Knoxville TN and Gastonia NC From kosmicdog <@t> hotmail.com Thu Oct 7 13:24:19 2004 From: kosmicdog <@t> hotmail.com (jason m) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] anhydrous EtOH for laser capture? Message-ID: Simple question...does anyone know of a good source for anhydrous EtOH for use in LCM? Preferably a company that will ship to Montréal. Or is there somewhere to purchase the "hydroscopic-beads" which can dry EtOH? Thanks, Jason. _________________________________________________________________ Take charge with a pop-up guard built on patented Microsoft® SmartScreen Technology http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines Start enjoying all the benefits of MSN® Premium right now and get the first two months FREE*. From darrenj <@t> medica.co.nz Thu Oct 7 14:40:24 2004 From: darrenj <@t> medica.co.nz (Darren James) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Eye Protection Question In-Reply-To: Message-ID: <000901c4aca5$7d2e1590$c864a8c0@medica.co.nz> We used weak hypochlorite (ie Milton tablets), rinsed thoroughly after cleaning of course. Darren James -----Original Message----- From: Molinari, Betsy [mailto:BMolinari@heart.thi.tmc.edu] Sent: Friday, 8 October 2004 12:10 a.m. To: darrenj@medica.co.nz Subject: RE: [Histonet] Eye Protection Question Our stations get flushed regularly also but the idea of doing cultures is one I would like to start. How did you clean the dirty stations. Betsy Molinari HT(ASCP) Texas Heart Institute Houston,Texas 77030 832-355-6524 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Darren James Sent: Wednesday, October 06, 2004 2:58 PM To: 'Joe Nocito'; Histonet (E-mail) Subject: RE: [Histonet] Eye Protection Question Hi, I agree totally with Joe's point about flushing your eyewash stations regularly. These are often forgotten about and can become incredibly contaminated over a very short period. We had micro prepare some plates on the water in the eyewash stations and there were some fairly pathogenic species present. Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joe Nocito Sent: Thursday, 7 October 2004 5:00 a.m. To: Jordan Brod; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Eye Protection Question Jordon, we have full-face safety shields as well as surgical masks with an attached eye shield. We wear either when changing the staining machines and tissue processors. When I gross, I wear my eyeglasses that are classified as safety wear. We have never had an incident, but we flush our eyewash stations weekly because you never know what will happen. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jordan Brod Sent: Wednesday, October 06, 2004 8:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Eye Protection Question Good Morning! What kind of eye protection are technicians required to wear in the laboratory during grossing, sectioning, processor maintenance, etc.? Thanks and have a great day. Jordan TVMDL - College Station, TX _______________________________________________ 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 From gentras <@t> vetmed.auburn.edu Thu Oct 7 14:48:14 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:24:08 2005 Subject: Fwd: Re: Fwd: Re: [Histonet] mouse brains Message-ID: <6.0.1.1.0.20041007144800.01b55ca8@mailhost.vetmed.auburn.edu> >Date: Thu, 07 Oct 2004 14:47:42 -0500 >To: LuAnn Anderson >From: "Atoska S. Gentry" >Subject: Re: Fwd: Re: [Histonet] mouse brains > > >Please accept my apology, no offense intended. My original inquiry was the >maximum amount of time one can hold mouse brain in PFA.. You're exactly >correct in regards to the sectioning of animal tissues, but human tissues >sectioned better when chilled 23+ years ago when I was in that >discipline. Sounds like your tissues are undergoing extreme dehydration >and you're having to rehydrate in order to get sections. Thanks again. Atoska > >At 01:22 PM 10/7/2004, you wrote: >>I've been doing this for 20 years--it ALWAYS works. Wild type or knock >>out, makes no difference. You put them on ice after soaking to cool them >>down at that point. I do this on a daily basis and have never had >>problems or come across blocks that require complete cold at all times??? >>This does not make sense to me. Anyway, This is what works for me and I >>get beautiful sections---first time, every time. If you don''t want to >>try it that's fine. It was just a suggestion. >> >> >>At 12:20 PM 10/7/04, you wrote: >> >>>Thanks! yes, I'm familiar with that procedure. I used it for the knock >>>out brains and it worked pretty good, but the wild type are requiring >>>complete cold. Fortunately, as mentioned through trial & error I've been >>>able to obtain sections. I'm just curious about avenues to avoid for >>>mouse brains to be sectioned beyond this point. Atoska >>> >>>At 11:46 AM 10/7/2004, you wrote: >>>>Face blocks and soak face down in warm water or warm water with a drop >>>>if dish soap. Soak for at least 20 minutes (or longer). Place blocks on >>>>ice to cool. Sectioning should be easy after this. In 20 years, this >>>>method has worked flawlessly every time with animal brain sections. >>>> >>>> >>>>At 10:26 AM 10/7/04, you wrote: >>>> >>>>>>Date: Thu, 07 Oct 2004 10:26:10 -0500 >>>>>>To: Gayle Callis >>>>>>From: "Atoska S. Gentry" >>>>>>Subject: Re: [Histonet] mouse brains >>>>>> >>>>>> >>>>>>Gayle, thanks for your reply. Actually, I'm processing these samples >>>>>>for a professor from the Biological Science Department on main >>>>>>campus. And she's conducting various IHC stains for "Tau Filament >>>>>>Formation in Transgenic Mice Expressing P301L Tau" & neurofibrillary >>>>>>changes in CNS. She's working in conjunction with some Spanish >>>>>>Scientists. I believe the samples were shipped from Spain in PFA back >>>>>>in February 04'. When we receive them they are whole brains in >>>>>>PFA from which coronal slices are taken, processed, and sectioned. >>>>>>The first few sets of brain received a few months ago sectioned much >>>>>>easier, but the two she brought out a couple of weeks ago are >>>>>>requiring a fair amount of trial & error to obtain fairly decent >>>>>>sections. Atoska >>>>>> >>>>>> >>>>>>At 03:11 PM 10/6/2004, you wrote: >>>>>>>Atoska, >>>>>>> >>>>>>>More info please. Are you processing brain slices cut with a matrix >>>>>>>device or whole brain. We do both and find the time in processing >>>>>>>is critical. We just did whole mouse brains fixed for three weeks >>>>>>>in NBF for LFB staining and H&E and had wonderful sections but used >>>>>>>a longer processing schedule than for our coronal mouse brain slices. >>>>>>> >>>>>>>For our coronal sections, we perfuse after anesthetizing animal - >>>>>>>first with saline followed by PLP via heart (a morning protocol then >>>>>>>fix for an additonal 5 hours, slice into 3 mm thick slices with >>>>>>>matrix device prior to a special mouse brain processing schedule on >>>>>>>a VIP the same evening. PLP has paraformaldehyde so will be similar >>>>>>>to your PFA fixation and are able to obtain excellent coronal >>>>>>>sections. I have not noticed any differences between longer NBF nor >>>>>>>PLP fixation on sectioning effects. >>>>>>> >>>>>>>If we fix brain with PFA, we do this overnight at 4C, and if the >>>>>>>fixation needs to be longer, we change the PFA to fresh. Are you >>>>>>>perfusing (ideal situation) then immersing samples overnight, >>>>>>>immersion only, at RT or 4C. >>>>>>> >>>>>>>What sectioning problems do you encounter? >>>>>>> >>>>>>> >>>>>>> >>>>>>>At 01:08 PM 10/6/2004, you wrote: >>>>>>> >>>>>>>>Hello, those of you processing & sectioning coronal sections of >>>>>>>>mouse brain; from you experience can prolonged fixation in >>>>>>>>4%PFA be the culprit of sectioning problems? What is the maximum >>>>>>>>fixation time period the brain should held in PFA before processing >>>>>>>>& sectioning? Thanks! Atoska >>>>>>>> >>>>>>>> >>>>>>>>Atoska S. Gentry B.S., HT(ASCP) >>>>>>>>Research Assistant III >>>>>>>>Scott-Ritchey Research Center >>>>>>>>College of Veterinary Medicine >>>>>>>>Auburn University, AL 36849 >>>>>>>>Phone# (334)844-5579 Fax# (334)844-5850 >>>>>>>> >>>>>>>>_______________________________________________ >>>>>>>>Histonet mailing list >>>>>>>>Histonet@lists.utsouthwestern.edu >>>>>>>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>>>>>> >>>>>>>Gayle Callis >>>>>>>MT,HT,HTL(ASCP) >>>>>>>Research Histopathology Supervisor >>>>>>>Veterinary Molecular Biology >>>>>>>Montana State University - Bozeman >>>>>>>PO Box 173610 >>>>>>>Bozeman MT 59717-3610 >>>>>>>406 994-6367 (lab with voice mail) >>>>>>>406 994-4303 (FAX) >>>>> >>>>> >>>>>_______________________________________________ >>>>>Histonet mailing list >>>>>Histonet@lists.utsouthwestern.edu >>>>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gentras <@t> vetmed.auburn.edu Thu Oct 7 14:56:41 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:24:08 2005 Subject: Fwd: Re: [Histonet] mouse brains Message-ID: <6.0.1.1.0.20041007145233.01b55ca8@mailhost.vetmed.auburn.edu> Thanks so much. You're exactly correct. Our in house fixation in 4%PFA is done strictly at 4C overnight, but we've not done mouse brains before. Therefore, I had to devise a whole new processing schedule for them. Atoska >X-Sender: gcallis@gemini.msu.montana.edu >X-Mailer: QUALCOMM Windows Eudora Version 6.0.0.22 >Date: Thu, 07 Oct 2004 13:20:57 -0600 >To: "Atoska S. Gentry" >From: Gayle Callis >Subject: Re: [Histonet] mouse brains >X-spam: ESP<2>=RBL:<0> RDNS:<0> SHA:<2> UHA:<0> SLS:<0> BAYES:<0> SPF:<0> >Porn Dictionary (TRU5):<0> Spam Dictionary (TRU5):<0> HTML Dictionary >(TRU5):<0> DRUGS2:<0> Obscenities Dictionary (TRU5):<0> CAN-SPAM >Compliance Dictionary (TRU5):<0> NigeriaScam Dictionary (TRU5):<0> Embed >HTML Dictionary (TRU5):<0> URL Dictionary (TRU5):<0> porn2:<0> > >Atoska, > >For one thing, PFA is NOT stable, it tends to break down, go bad far too >fast. The brains should have been totally fixed on site, then transferred >to either 70% ethanol, even 50% alcohol after rinsing with PBS and shipped >that way. RT fixation with PFA has always been a no no, guess left over >from EM days, but everyone I work with doing PFA fixation does the >fixation at 4C, then proceeds from there. I suggest they suspend (if they >do immersion fixation) the brains, hang them so brains are not sitting on >bottom of a container to allow all sides/surfaces to have contact with >PFA, particularly if they do immersion fixation. I would hope they start >doing perfusion followed by immersion, far better with brain and PFA. > >We would never store brains in PFA, rather do fixation and transfer to >alcohol for storage, then process. If we go longer than overnight in PFA, >the fixative is changed to fresh just because it is unstable. We make >ours up in Dulbeccos PBS, and never exceed 60C during PFA solution >preparation. Always far more difficult to deal with other peoples problems!!! > >Gayle Callis > > >At 09:26 AM 10/7/2004, you wrote: > >>Gayle, thanks for your reply. Actually, I'm processing these samples for >>a professor from the Biological Science Department on main campus. And >>she's conducting various IHC stains for "Tau Filament Formation in >>Transgenic Mice Expressing P301L Tau" & neurofibrillary changes in CNS. >>She's working in conjunction with some Spanish Scientists. I believe the >>samples were shipped from Spain in PFA back in February 04'. When we >>receive them they are whole brains in PFA from which coronal slices are >>taken, processed, and sectioned. The first few sets of brain received a >>few months ago sectioned much easier, but the two she brought out a >>couple of weeks ago are requiring a fair amount of trial & error to >>obtain fairly decent sections. Atoska >> >> >>At 03:11 PM 10/6/2004, you wrote: >>>Atoska, >>> >>>More info please. Are you processing brain slices cut with a matrix >>>device or whole brain. We do both and find the time in processing is >>>critical. We just did whole mouse brains fixed for three weeks in NBF >>>for LFB staining and H&E and had wonderful sections but used a longer >>>processing schedule than for our coronal mouse brain slices. >>> >>>For our coronal sections, we perfuse after anesthetizing animal - first >>>with saline followed by PLP via heart (a morning protocol then fix for >>>an additonal 5 hours, slice into 3 mm thick slices with matrix device >>>prior to a special mouse brain processing schedule on a VIP the same >>>evening. PLP has paraformaldehyde so will be similar to your PFA >>>fixation and are able to obtain excellent coronal sections. I have not >>>noticed any differences between longer NBF nor PLP fixation on >>>sectioning effects. >>> >>>If we fix brain with PFA, we do this overnight at 4C, and if the >>>fixation needs to be longer, we change the PFA to fresh. Are you >>>perfusing (ideal situation) then immersing samples overnight, immersion >>>only, at RT or 4C. >>> >>>What sectioning problems do you encounter? >>> >>> >>> >>>At 01:08 PM 10/6/2004, you wrote: >>> >>>>Hello, those of you processing & sectioning coronal sections of mouse >>>>brain; from you experience can prolonged fixation in 4%PFA be the >>>>culprit of sectioning problems? What is the maximum fixation time >>>>period the brain should held in PFA before processing & sectioning? >>>>Thanks! Atoska >>>> >>>> >>>>Atoska S. Gentry B.S., HT(ASCP) >>>>Research Assistant III >>>>Scott-Ritchey Research Center >>>>College of Veterinary Medicine >>>>Auburn University, AL 36849 >>>>Phone# (334)844-5579 Fax# (334)844-5850 >>>> >>>>_______________________________________________ >>>>Histonet mailing list >>>>Histonet@lists.utsouthwestern.edu >>>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>> >>>Gayle Callis >>>MT,HT,HTL(ASCP) >>>Research Histopathology Supervisor >>>Veterinary Molecular Biology >>>Montana State University - Bozeman >>>PO Box 173610 >>>Bozeman MT 59717-3610 >>>406 994-6367 (lab with voice mail) >>>406 994-4303 (FAX) From GauchV <@t> mail.amc.edu Thu Oct 7 15:04:02 2004 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] IMF Tissue Message-ID: I am posting this for one of our tech specialists here....Is anyone out there using anything to to mark tissue that is being cut for immunofluorescence in order to make it stand out against the OCT during cutting? Currently some of the renal biopsies are so thin that once they are rinsed they are very white and difficult to see when cutting. We tried using the colored OCT which did help a little but she is looking for something which will make it even better since these biopsies are so tiny and there is no tissue to spare. Thanks in advance for any help you can give me.... Have a great day !!! Vicki Gauch AMCH Albany, NY From lleach <@t> uic.edu Thu Oct 7 15:37:12 2004 From: lleach <@t> uic.edu (Lu Leach) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Collagen IV antibody Message-ID: <6.1.0.6.2.20041007153416.02479828@tigger.uic.edu> Dear Histonetters, Can anyone tell me where to acquire Collagen IV A5 antibody? Thanks, Lu Leach University of Illinois at Chicago Research Pathology Core 312-996-3869 From jrutherford <@t> sfbr.org Thu Oct 7 16:47:41 2004 From: jrutherford <@t> sfbr.org (Julienne Rutherford) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] placental IHC for the novice Message-ID: Hello. I'm new to histonet and am not trained in histo or IHC, but am very interested. I'm throwing myself into the fray, and hope you can help me. In response, which I welcome privately or in the forum, please use simple language for I am but a simple woman with a simple dream of studying monkey placentas. I want to study IGF-1 localization in the marmoset monkey placenta. I'm doing a lot of reading regarding methods, but I need help as to what I need to do (and by extension, how to ask for the money to do it in grant applications!). The folks in our histo lab are going to help me and train me but I want to come to them more prepared and I have to provide the specific materials I'll need. I have both 10% NBF-fixed tissues (most sitting around in formalin for a very long time), and I also have frozen, but by this I mean stuck in the regular old lab freezer to freeze conventionally. I will begin snap-freezing placentae we collect from now on, but what can I get out of this old stuff? Can I thaw out what I have and snap-freeze them in OCT? Should I try to do IHC on both the formalin-fixed AND the frozen tissue? In order to build a budget I need to know everything I need to make these slides. So, what kind of slides and coverslips do I need, what are all the chemicals involved, what is a secondary antibody and why do I need it, is a chromagen different from a stain, etc.? Also, is there something akin to and "IHC for Dummies" resource? I've looked at ihcworld.com for protocols but it's over my head at this point. I recognize that this is a skill perfected over many years, but I really would like a primer on the basics so I can have a more informed discussion about it. If the elementary pitch of my questions is inappropriate to the forum, I sincerely apologize. I'm just not sure where else to turn. Please respond to my Texas e-mail if you'd prefer to school (or scold) me privately! Thanks in advance, Julienne Rutherford Research Assistant (Adjunct) Southwest Foundation for Biomedical Research Southwest National Primate Research Center P.O. Box 760549 San Antonio, Texas 78245-0549 phone (210) 258-9864 fax (210) 258-9883 jrutherford@icarus.sfbr.org Ph.D. candidate Indiana University Bloomington, Indiana jnruther@indiana.edu From Jacqueline.Miller <@t> UTSouthwestern.edu Thu Oct 7 17:12:46 2004 From: Jacqueline.Miller <@t> UTSouthwestern.edu (Jacqueline Miller) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] anhydrous EtOH for laser capture? Message-ID: We have some that came as a kit, but they may sell it individually. It's sold by Arcturus. Catalog #KIT0411. 1-888-446-7911 650-962-3020 www.arctur.com Don't know if they will ship to Montreal, but they might be able to tell you someone who can. Jackie Miller >>> "jason m" 10/07/04 1:24 PM >>> Simple question...does anyone know of a good source for anhydrous EtOH for use in LCM? Preferably a company that will ship to Montr?al. Or is there somewhere to purchase the "hydroscopic-beads" which can dry EtOH? Thanks, Jason. _________________________________________________________________ Take charge with a pop-up guard built on patented Microsoft? SmartScreen Technology http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines Start enjoying all the benefits of MSN? Premium right now and get the first two months FREE*. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From darkdaym <@t> mindspring.com Thu Oct 7 18:11:30 2004 From: darkdaym <@t> mindspring.com (Mark Ray) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] Re: GI bx In-Reply-To: <06609E92.2D708D4B.0BF1BB09@aol.com> References: <06609E92.2D708D4B.0BF1BB09@aol.com> Message-ID: <4165CD22.3040504@mindspring.com> I have to agree with Bob. E K Industries has been selling Reagent Alcohol to clinical and research labs for quite a while. Longer than I care to think about. Never had a complaint and never heard anything like this. A while back, someone on Histonet explained how the combination of the two denaturing agents produced a mixture indistinguishable in its properties from absolute ethanol in histology applications. I can't remember who or when. Any other Reagent Alcohol users care to comment? Mark Ray E K Industries RSRICHMOND@aol.com wrote: >In regard to alcohols: > >Do people have really strong evidence that denatured alcohol behaves any differently from pure ethanol? > >Absolute (100%) ethanol is a major security problem for the laboratory, since tax authorities (Bureau of Alcohol, Tobacco, and Firearms - Department of Justice - in the USA) want to make really sure that it isn't diverted for beverage use. Many hospitals just don't have the storage facilities. > >In common use is what's generically called "reagent alcohol", with 90% ethanol, 5% methanol, and 5% isopropanol. This is denaturation formula "S3DA modified", the "modified" referring to the fact that isopropanol is also added to the methanol-denatured S3DA mix. - Another denaturant permitted in histology is methylisobutylketone (MIBK), which smells so bad that most labs don't want to use it. (Alcohol denatured with acetone is also available - unacceptable because it removes eosin instantly.) > >This old pathologist hasn't seen any difference between pure ethanol and reagent alcohol. Is any of you sure that there is a difference? > >Bob Richmond >Samurai Pathologist >Knoxville TN and Gastonia NC > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From aldo.anile <@t> HDScientific.com.au Thu Oct 7 18:18:34 2004 From: aldo.anile <@t> HDScientific.com.au (Aldo Anile) Date: Fri Sep 16 15:24:08 2005 Subject: [Histonet] RE: Tissue Processors Message-ID: The Medite TPC15 Tissue Processor has a tissue transfer system similar to an x-y staining machine. The advantage of such a system is that you can run two similar or different processing schedules simultaneously. Effectively you have two processors for the price of one. Vaccuum and pressure can still be applied during processing. The other benefit is that there is no retort system present that relies on pumps, solenoids or clean lines. Aldo Anile Applications Consultant HD SCIENTIFIC SUPPLIES PTY LTD E-mail: aldo.anile@hdscientific.com.au Web: www.hdscientific.com.au From TJasper <@t> smdc.org Thu Oct 7 18:53:35 2004 From: TJasper <@t> smdc.org (Jasper, Thomas G.) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Kermit Groothuis Memorial Scholarship Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA02E44FC6@harrier> Dear Vicki and friends of Kermit, I am saddened to hear that Kermit has passed away. I first met Kermit about 20 years ago in Madison. You knew immediately that he was a good soul. As I moved on in my career I lost track of Kermit and then began running into him at NSH conventions. I last saw him in Louisville and he always genuinely glad to see you. It probably didn't hurt any that I was/am a die-hard Badger fan. I'm glad that Bucky showed up at his retirement. Although I was not a close acquaintance I certainly feel fortunate to have known Kermit. Perhaps (Vicki) you would pass my condolences to his family. Kermit really was one of the "good guys". Thank you, Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: Vicki Kalscheur [mailto:kalschev@svm.vetmed.wisc.edu] Sent: Thursday, October 07, 2004 1:17 PM To: Histonet Discussion Subject: [Histonet] Kermit Groothuis Memorial Scholarship Kermit Groothuis Honored at Retirement Reception Kermit Groothuis, an 18-year veteran at the University of Wisconsin-Madison School of Veterinary Medicine, retired from his position in histopathology in August. Even Bucky Badger, the University of Wisconsin mascot, came to share in the celebration. Over the years, Kermit made numerous friends at the SVM as well as state and national histology forums. In recognition, for his service, the Wisconsin Histotechnology Society and the National Society for Histotechnology, as well as colleagues from his labs purchased several SVM tiles in his name, honoring his many contributions. These tiles have been installed in front of the University of Wisconsin - School of Veterinary Medicine. A good time was had by all at the retirement reception. Regretfully, our friend and colleague, Kermit B. Groothuis, (HT. ASCP.), 64 years young, passed away after a short bout with cancer. A lover of all sports, he refereed team sports and worked UW football and basketball games, and was also active in the Boy Scouts. He enjoyed all people and was a friend to all. Our heartfelt condolences go out to his wife, Mary, and family members. Kermit B. Groothuis September 25, 1939-September 21, 2004 God Bless For interested parties, colleagues and friends: A Kermit Groothuis Memorial Scholarship is being established, for a veterinary student, to be awarded, each year at a graduation banquet. If you wish to contribute make checks payable to: University of Wisconsin Foundation. Mail to Att: Nancy J Nelson, School of Veterinary Medicine, 2015 Linden Dr. Madison, WI 53706. Or visit www.vetmed.wisc.edu to give online. Please specify the Kermit Groothuis Memorial Scholarship. Email questions to nelson@svm.vetmed.wisc.edu From kcboswell <@t> grandecom.net Thu Oct 7 20:18:32 2004 From: kcboswell <@t> grandecom.net (Boswell) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] immunostaining Message-ID: <002101c4acd4$bd5672a0$e1edbcd8@Boswell> If our lab wanted to start learning and doing immunostaining first by hand then with strainer where would we start. Are there any textbooks out there. No one in our lab has ever been around immuno's and there is not a lab close by that we could learn from. Please help!!!! Thank you, Chantel From kosmicdog <@t> hotmail.com Thu Oct 7 20:45:03 2004 From: kosmicdog <@t> hotmail.com (jason m) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] anhydrous EtOH clarification. Message-ID: For laser capture the EtOH must remain 100% dry. Using a bottle of regular anhydrous EtOH after it has been opened once or twice sometimes has absorbed enough moisture from the air to cause interferance with the LCM film and negatively affect the capture. I know that arcturus sells EtOH which has small beads mopping up any moisture contaimination. I just don't want to keep buying a whole 'kit' just for 500ml's of 100% EtOH. The kit is certified RNase free and works very well but not econimal for our purposes. The only other source i found for this type of material no longer ships to Canada. Hopefully that clears things up a little bit. As for those who wrote to me explaining that 'all' EtOH is anhydrous, please excuse me for assuming that you were familiar with problems facing laser capture as I am sure you would have better understood the nature of my problem. Thanks again for all the help. I have learned a lot from histonet over the last year, it is an excellent resource. aurevoir, jason. _________________________________________________________________ Designer Mail isn't just fun to send, it's fun to receive. Use special stationery, fonts and colors. http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines Start enjoying all the benefits of MSN® Premium right now and get the first two months FREE*. From pengbw <@t> sjtu.edu.cn Fri Oct 8 02:44:46 2004 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Smears Message-ID: <20041008074446.44D8710D2A1C@sjtu.edu.cn> Dear all histoneters, Are there anyone can help me out of the smears staining problem. I\'m doing lymphocyte smears staining these days as following: smears were made from lymphocyte which were in 1640. After air dry, smears were dip into methanol for several times and were stained in Giemsa satin to indetify macrophages. The result shows that almost holf of the cells in the smears have broken and leak out their cytosol and nucleus contents. What I have done wrong and what can I prevent the cells from broken? Yours sincerely, Baowei Peng Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China From ajennings <@t> unmc.edu Fri Oct 8 08:24:20 2004 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Vendors please In-Reply-To: <007401c4ac4e$9c84ec40$78e0d445@domainnotset.invalid> Message-ID: I left my email address with many of the vendors at NSH this year and I appreciate all of the information that I have received and anticipate receiving. I would like to ask those that send information to multiple listing please either use bcc or create an address list that does not share my email address with all of the people that you send it to. The address I left with you for information on your products is my work email and it is just a matter of nettiquette to not circulate it. Thank you From peoshel <@t> wisc.edu Fri Oct 8 08:44:43 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] anhydrous EtOH clarification. In-Reply-To: References: Message-ID: Jason, The beads are most likely either silica gel or molecular sieve. I don't know of specific sources in Canada, but cases of silica-gel beads in little capsules for drying electronics and pharmaceuticals can be gotten relatively cheaply from suppliers for those industries. I'm trying to track down such a supplier now that someone else here used, and if I get the information, I'll pass it on. Molecular sieves are better, in that they have a greater capacity for H20. We use them routinely to keep are EtOH dry for electron microscopy preparation. Molecular sieves can be purchased from industrial gas suppliers -- they're typically used for drying gases. Shop around, though, the prices can vary by a factor of 2 or more. You want 4 Angstrom beads -- the pore size that traps H20 -- and preferably with some indicator beads. Get the small size beads for greater surface area. Be sure to pipette from the top of the liquid if you use molecular sieves to avoid dust, or put the sieves in dialysis tubing. You should also be able to get these from electron microscopy supply houses, but for higher prices. It's worth while doing some shopping on the internet. Phil >For laser capture the EtOH must remain 100% dry. Using a bottle of >regular anhydrous EtOH after it has been opened once or twice >sometimes has absorbed enough moisture from the air to cause >interferance with the LCM film and negatively affect the capture. > >I know that arcturus sells EtOH which has small beads mopping up any >moisture contaimination. I just don't want to keep buying a whole >'kit' just for 500ml's of 100% EtOH. The kit is certified RNase free >and works very well but not econimal for our purposes. The only >other source i found for this type of material no longer ships to >Canada. > >Hopefully that clears things up a little bit. As for those who wrote >to me explaining that 'all' EtOH is anhydrous, please excuse me for >assuming that you were familiar with problems facing laser capture >as I am sure you would have better understood the nature of my >problem. > >Thanks again for all the help. I have learned a lot from histonet >over the last year, it is an excellent resource. >aurevoir, >jason. > >_________________________________________________________________ >Designer Mail isn't just fun to send, it's fun to receive. Use >special stationery, fonts and colors. >http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines >Start enjoying all the benefits of MSN? Premium right now and get >the first two months FREE*. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From georgecole <@t> ev1.net Fri Oct 8 09:57:33 2004 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Peggy Wenk, Vicki Gauch, Lynette Rlby, Peggy Lagaretta and Steve Joy Message-ID: <000001c4ad47$3082fab0$014dbad0@hppav> Peggy, Vicki, Lynette, Peggy and Steve; Your muscle and nerve packets should be arriving to you if they haven't arrived all ready. I hope they are of use to you. georgecole@ev1.net From gentras <@t> vetmed.auburn.edu Fri Oct 8 10:19:48 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:24:09 2005 Subject: Fwd: { SPAM 1 }::Re: Fwd: Re: [Histonet] mouse brains Message-ID: <6.0.1.1.0.20041008101927.01b43ad0@mailhost.vetmed.auburn.edu> Thanks!!! >X-Umn-Remote-Mta: [N] x141-96.psych.umn.edu [134.84.141.96] #+LO+HN >X-Sender: ander093@ander093.email.umn.edu >X-Mailer: QUALCOMM Windows Eudora Version 6.1.2.0 >Date: Thu, 07 Oct 2004 15:48:01 -0500 >To: "Atoska S. Gentry" >From: LuAnn Anderson >X-spam: ESP<53>=RBL:<0> RDNS:<0> SHA:<3> UHA:<0> SLS:<0> BAYES:<0> SPF:<0> >Porn Dictionary (TRU5):<0> Spam Dictionary (TRU5):<0> HTML Dictionary >(TRU5):<0> DRUGS2:<0> Obscenities Dictionary (TRU5):<50> CAN-SPAM >Compliance Dictionary (TRU5):<0> NigeriaScam Dictionary (TRU5):<0> Embed >HTML Dictionary (TRU5):<0> URL Dictionary (TRU5):<0> porn2:<0> >Subject: { SPAM 1 }::Re: Fwd: Re: [Histonet] mouse brains > >Aha!! The 4C is your problem. Cold slows down fixation immensely. It is an >"old wive's tale" that tissues must be fixed in the refrigerator with PFA. >Try fixing some at room temp and compare with some fixed at 4C. You will >get much better fixation and therefore better results at room temp. Hope >this helps. > >At 02:56 PM 10/7/04, you wrote: > >>Thanks so much. You're exactly correct. Our in house fixation in 4%PFA is >>done strictly at 4C overnight, but we've not done mouse brains >>before. Therefore, I had to devise a whole new processing schedule for >>them. Atoska >> >> >>>X-Sender: gcallis@gemini.msu.montana.edu >>>X-Mailer: QUALCOMM Windows Eudora Version 6.0.0.22 >>>Date: Thu, 07 Oct 2004 13:20:57 -0600 >>>To: "Atoska S. Gentry" >>>From: Gayle Callis >>>Subject: Re: [Histonet] mouse brains >>>X-spam: ESP<2>=RBL:<0> RDNS:<0> SHA:<2> UHA:<0> SLS:<0> BAYES:<0> >>>SPF:<0> Porn Dictionary (TRU5):<0> Spam Dictionary (TRU5):<0> HTML >>>Dictionary (TRU5):<0> DRUGS2:<0> Obscenities Dictionary (TRU5):<0> >>>CAN-SPAM Compliance Dictionary (TRU5):<0> NigeriaScam Dictionary >>>(TRU5):<0> Embed HTML Dictionary (TRU5):<0> URL Dictionary (TRU5):<0> porn2:<0> >>> >>>Atoska, >>> >>>For one thing, PFA is NOT stable, it tends to break down, go bad far too >>>fast. The brains should have been totally fixed on site, then >>>transferred to either 70% ethanol, even 50% alcohol after rinsing with >>>PBS and shipped that way. RT fixation with PFA has always been a no no, >>>guess left over from EM days, but everyone I work with doing PFA >>>fixation does the fixation at 4C, then proceeds from there. I suggest >>>they suspend (if they do immersion fixation) the brains, hang them so >>>brains are not sitting on bottom of a container to allow all >>>sides/surfaces to have contact with PFA, particularly if they do >>>immersion fixation. I would hope they start doing perfusion followed by >>>immersion, far better with brain and PFA. >>> >>>We would never store brains in PFA, rather do fixation and transfer to >>>alcohol for storage, then process. If we go longer than overnight in >>>PFA, the fixative is changed to fresh just because it is unstable. We >>>make ours up in Dulbeccos PBS, and never exceed 60C during PFA solution >>>preparation. Always far more difficult to deal with other peoples problems!!! >>> >>>Gayle Callis >>> >>> >>>At 09:26 AM 10/7/2004, you wrote: >>> >>>>Gayle, thanks for your reply. Actually, I'm processing these samples >>>>for a professor from the Biological Science Department on main campus. >>>>And she's conducting various IHC stains for "Tau Filament Formation in >>>>Transgenic Mice Expressing P301L Tau" & neurofibrillary changes in CNS. >>>>She's working in conjunction with some Spanish Scientists. I believe >>>>the samples were shipped from Spain in PFA back in February 04'. When >>>>we receive them they are whole brains in PFA from which coronal slices >>>>are taken, processed, and sectioned. The first few sets of brain >>>>received a few months ago sectioned much easier, but the two she >>>>brought out a couple of weeks ago are requiring a fair amount of trial >>>>& error to obtain fairly decent sections. Atoska >>>> >>>> >>>>At 03:11 PM 10/6/2004, you wrote: >>>>>Atoska, >>>>> >>>>>More info please. Are you processing brain slices cut with a matrix >>>>>device or whole brain. We do both and find the time in processing is >>>>>critical. We just did whole mouse brains fixed for three weeks in >>>>>NBF for LFB staining and H&E and had wonderful sections but used a >>>>>longer processing schedule than for our coronal mouse brain slices. >>>>> >>>>>For our coronal sections, we perfuse after anesthetizing animal - >>>>>first with saline followed by PLP via heart (a morning protocol then >>>>>fix for an additonal 5 hours, slice into 3 mm thick slices with matrix >>>>>device prior to a special mouse brain processing schedule on a VIP the >>>>>same evening. PLP has paraformaldehyde so will be similar to your PFA >>>>>fixation and are able to obtain excellent coronal sections. I have >>>>>not noticed any differences between longer NBF nor PLP fixation on >>>>>sectioning effects. >>>>> >>>>>If we fix brain with PFA, we do this overnight at 4C, and if the >>>>>fixation needs to be longer, we change the PFA to fresh. Are you >>>>>perfusing (ideal situation) then immersing samples overnight, >>>>>immersion only, at RT or 4C. >>>>> >>>>>What sectioning problems do you encounter? >>>>> >>>>> >>>>> >>>>>At 01:08 PM 10/6/2004, you wrote: >>>>> >>>>>>Hello, those of you processing & sectioning coronal sections of mouse >>>>>>brain; from you experience can prolonged fixation in 4%PFA be the >>>>>>culprit of sectioning problems? What is the maximum fixation time >>>>>>period the brain should held in PFA before processing & sectioning? >>>>>>Thanks! Atoska >>>>>> >>>>>> >>>>>>Atoska S. Gentry B.S., HT(ASCP) >>>>>>Research Assistant III >>>>>>Scott-Ritchey Research Center >>>>>>College of Veterinary Medicine >>>>>>Auburn University, AL 36849 >>>>>>Phone# (334)844-5579 Fax# (334)844-5850 >>>>>> >>>>>>_______________________________________________ >>>>>>Histonet mailing list >>>>>>Histonet@lists.utsouthwestern.edu >>>>>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>>>> >>>>>Gayle Callis >>>>>MT,HT,HTL(ASCP) >>>>>Research Histopathology Supervisor >>>>>Veterinary Molecular Biology >>>>>Montana State University - Bozeman >>>>>PO Box 173610 >>>>>Bozeman MT 59717-3610 >>>>>406 994-6367 (lab with voice mail) >>>>>406 994-4303 (FAX) >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet From georgecole <@t> ev1.net Fri Oct 8 10:25:45 2004 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Muscle and Nerve packet Message-ID: <000001c4ad4b$1856ca30$094dbad0@hppav> Floyd; Your muscle and nerve packet was returned to me in the mail. Is there something wrong with the address I sent it to?: I mailed it 8/24/04 to: FLOYD GRIMM III INDIANA UNIVERSITY SCHOOL OF MEDICINE COLEMAN HALL ROOM 322 1140 WEST MICHIGAN STREET IMDIANAPOLIS. IN 46202-5113 From BWinters <@t> NCH.ORG Fri Oct 8 11:04:38 2004 From: BWinters <@t> NCH.ORG (Winters, Bert) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] PROFICENCY TRAINING IN HISTOLOGY Message-ID: <270614B321ACB44D8C1D91F4F921FDC3627782@NCH01EX02.nch.org> For histotech we have always had a competency check list but have never had a preficiency test per se. Can anyone out there give me a example of what they would include or how to run proficency test for histotechnolgists. Bert Winters HTL (ASCP) NORTHWEST COMMUNITY HOSPITAL ARLINGTON HEIGHTS, ILL EMAIL-----BWINTERS@NCH.ORG ******************* PLEASE NOTE ******************* This E-Mail/telefax message and any documents accompanying this transmission may contain information that is privileged, confidential, and/or exempt from disclosure under applicable law and is intended solely for the addressee(s) named above. If you are not the intended addressee/recipient, you are hereby notified that any use of, disclosure, copying, distribution, or reliance on the contents of this E-Mail/telefax information is strictly prohibited and may result in legal action against you. Please reply to the sender advising of the error in transmission and immediately delete/destroy the message and any accompanying documents. Thank you. From gentras <@t> vetmed.auburn.edu Fri Oct 8 11:22:27 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] LuAnn Anderson Message-ID: <6.0.1.1.0.20041008111911.01affd98@mailhost.vetmed.auburn.edu> LuAnn, good morning! I really appreciate all of your pointers on mouse brain sectioning, and today I'm going to give your recommended protocol a try. I notice you said the faced blocks can soak at least 20 min. or longer, please I'm wondering do you chill for a specific amount of time? Is more chilled or less better in your experience? Thanks again. Atoska From SRenn <@t> CGR.Harvard.edu Fri Oct 8 11:35:23 2004 From: SRenn <@t> CGR.Harvard.edu (Susan Renn) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] cryostat brain in situ / tecan Message-ID: <339D68B133EAD311971E009027DC479701D489A0@montecarlo.cgr.harvard.edu> Hello I am looking for a good protocol for DIG labeled in situ on brain cryostat sections (from a fish). I am using a tecan liquid handler. The protocol that I have right now has a dehydration step which I am not familiar with from my previous work with in situ hyb in drosophila.... Is this step necessary for brain sections? What does it do? Also, I have seen protocols which call for fixation before sectioning and others that call for quick freezing in oct and fixation after sectioning. What are the pros and cons of these two techniques. Furhtermore, if one does fix after sectioning, should it be done immediately after sectioning, after the sections have warm dried for 20 min, or just before hybridization after the sections have been stored at -80 for some time? Any answers are greatly appreciated. Suzy From tmhpath <@t> amigo.net Fri Oct 8 17:53:40 2004 From: tmhpath <@t> amigo.net (Michelle D. Moore) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] tissue processors References: <20041007150612.52609.qmail@web61110.mail.yahoo.com> Message-ID: <001d01c4ad89$a8678d60$4000a8c0@ever> I recently purchaed a TBS ATP-1 processor and have had wonderful results from it. No problems currently. Runs well is quiet, and has been my life saver lately. I like all the features on it shows you when things have gone wrong and you can look back at the alarm archive to see if you have missed any alarms that weren't critical. If there is a problem and the process is interrupted it goes back to the last station it was on and will hold there til you can fix the problem. It has a special station to set up for reverse proceesing if you need it and you can set up many different processing schedules, times, vacumn on or off, you can set agitation times on all stations, and you can set different temperatures on each stations. I think it is a wonderful unit but that is just one opinion others may think differently. I hope this helps you with you decision. If you want to contact me off the histonet my e-mail address is tmhpath@amigo.net . Good luck with you decision. Michelle Moore HT(ASCP) The Memorial Hospital Craig, Colorado ----- Original Message ----- From: "esteban enriquez" To: "GUTIERREZ, JUAN" ; "Andres Kulla" ; Sent: Thursday, October 07, 2004 9:06 AM Subject: RE: [Histonet] tissue processors > Tissue Tek VIPs is reliable, they don't has the problems with line clogging continually like some Company's Polaris tissue processor. What the good of higher throughput if the thing is always needing to be fixed??? I stay with the Tissue Tek. > > EE > > "GUTIERREZ, JUAN" wrote: > Nothing beats the Tissue Teks. They keep going and going and going. Sorry but I can not say anything bad about equipment or companies anymore. Too many damn lawyers here in the USofA. > > Juan > > -----Original Message----- > From: Andres Kulla [mailto:Andres.Kulla@kliinikum.ee] > Sent: Tuesday, October 05, 2004 1:11 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] tissue processors > > To the experts in automated tissue processing, > Could anybody share his/her experience with TBS (Triangle Biomedical Sciences) ATP1 tissue processor. Positive vs negative aspects in comparison with other devices eg Sakuras Tissue Tek VIP 500. > With best regards from > Andres Kulla, > Estonia > > _______________________________________________ > 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 > > > --------------------------------- > Do you Yahoo!? > vote.yahoo.com - Register online to vote today! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From CrochiereSteve <@t> aol.com Fri Oct 8 19:48:29 2004 From: CrochiereSteve <@t> aol.com (CrochiereSteve@aol.com) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Proficiency Testing Message-ID: Hi, I subscribe to the Histology QIP program. One receives a list of specimens to cut and stain, then submits the slides to CAP for "grading" Steven M. Crochiere, HT(ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center Springfield, MA 01104 From mariatere <@t> infovia.com.ar Sat Oct 9 05:10:44 2004 From: mariatere <@t> infovia.com.ar (Teresa Dominguez) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Smears References: <20041008074446.44D8710D2A1C@sjtu.edu.cn> Message-ID: <01fd01c4ade8$3fd61870$f77c46c8@DFI> What I do with old smears is : To place the smears in a Fisiologic solution for , at least 10 minutes, and then re-fixation in 95% alcohol, after that you can stian the smean with the technicque you want. Good luck! Ht. Maria Teresa Dominguez Patology Service Rio Grande Reg. Hospital Rio Grande, Tierra del Fuego, Argentina. ----- Original Message ----- From: "Baowei Peng" To: Sent: Friday, October 08, 2004 4:44 AM Subject: [Histonet] Smears > Dear all histoneters, > > Are there anyone can help me out of the smears staining problem. > I\'m doing lymphocyte smears staining these days as following: > smears were made from lymphocyte which were in 1640. After air dry, smears were dip into methanol for several times and were stained in Giemsa satin to indetify macrophages. The result shows that almost holf of the cells in the smears have broken and leak out their cytosol and nucleus contents. > What I have done wrong and what can I prevent the cells from broken? > Yours sincerely, > > > > Baowei Peng > Pharmacy School > Shanghai Jiaotong University > Shanghai, 200030 > China > ---------------------------------------------------------------------------- ---- > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From amosbrooks <@t> earthlink.net Sat Oct 9 10:40:53 2004 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] General IHC In-Reply-To: <200410080936.1cfXJ31QJ3NZFmQ0@tanager.mail.pas.earthlink.n et> References: <200410080936.1cfXJ31QJ3NZFmQ0@tanager.mail.pas.earthlink.net> Message-ID: <6.0.0.22.0.20041009110321.01c734a0@pop.earthlink.net> Chantel & Julienne, Both of these questions are very similar. Likewise for both of you I would suggest that you read a publication by DAKO called Basic Immunohistochemistry, you can find them at www.dakocytomation.com it is a very easy read and is the best crash course that I've seen. It will give you an overview that should give you enough information to get you started. Lawrence True also has a book (by the same title I think) that is excellent as well. (Check with Amazon.com for this) You really should know as much about the how and why as possible when doing IHC as if anything goes wrong in the process you'll know how to fix it. Don't let this intimidate you as most IHC can be as easy or as complex as you want it to be. With most automatic stainers you can train a monkey to use them. I don't recommend it for the above reasons but it can be done. Generally animal IHC is more difficult as diagnostic IHC usually is geared toward humans. I mostly work with humans so I can only suggest sources of information for animal work. For example I know DAKO has a secondary detection kit specifically for animals. There are other vendors as well I am sure any of them would love to give you more information. Fixation is very important and will radically change the whole process. Formalin fixed tissues usually require some step to (for lack of a better term) undo the fixation. So, Julienne, you will find there will be a difference in the fixed placentas and the frozen ones. I suggest that you figure out what antibodies you are going to use then determine a vendor from there (usually base this decision on price and availability). Then contact a sales rep and ask them to guide you thru the process. Most vendors will have very good tech support. (DAKO, Ventana, BioCare etc) Some offer classes and tutorials to really train you in how things work. DAKO actually has workshops in California that are very good. Good Luck, Amos Brooks At 12:36 PM 10/8/2004, you wrote: >Message: 10 >Date: Thu, 7 Oct 2004 16:47:41 -0500 >From: "Julienne Rutherford" >Subject: [Histonet] placental IHC for the novice >To: "Histonet@Lists. Utsouthwestern. Edu" > >Message-ID: >Content-Type: text/plain; charset="iso-8859-1" > >Hello. I'm new to histonet and am not trained in histo or IHC, but am very >interested. I'm throwing myself into the fray, and hope you can help me. In >response, which I welcome privately or in the forum, please use simple >language for I am but a simple woman with a simple dream of studying monkey >placentas. I want to study IGF-1 localization in the marmoset monkey >placenta. I'm doing a lot of reading regarding methods, but I need help as >to what I need to do (and by extension, how to ask for the money to do it in >grant applications!). The folks in our histo lab are going to help me and >train me but I want to come to them more prepared and I have to provide the >specific materials I'll need. I have both 10% NBF-fixed tissues (most >sitting around in formalin for a very long time), and I also have frozen, >but by this I mean stuck in the regular old lab freezer to freeze >conventionally. I will begin snap-freezing placentae we collect from now on, >but what can I get out of this old stuff? Can I thaw out what I have and >snap-freeze them in OCT? Should I try to do IHC on both the formalin-fixed >AND the frozen tissue? > >In order to build a budget I need to know everything I need to make these >slides. So, what kind of slides and coverslips do I need, what are all the >chemicals involved, what is a secondary antibody and why do I need it, is a >chromagen different from a stain, etc.? Also, is there something akin to and >"IHC for Dummies" resource? I've looked at ihcworld.com for protocols but >it's over my head at this point. I recognize that this is a skill perfected >over many years, but I really would like a primer on the basics so I can >have a more informed discussion about it. > >If the elementary pitch of my questions is inappropriate to the forum, I >sincerely apologize. I'm just not sure where else to turn. Please respond to >my Texas e-mail if you'd prefer to school (or scold) me privately! > >Thanks in advance, >Julienne Rutherford > >Research Assistant (Adjunct) >Southwest Foundation for Biomedical Research >Southwest National Primate Research Center >P.O. Box 760549 >San Antonio, Texas 78245-0549 >phone (210) 258-9864 >fax (210) 258-9883 >jrutherford@icarus.sfbr.org > >Ph.D. candidate >Indiana University >Bloomington, Indiana >jnruther@indiana.edu From JKOBLER <@t> PARTNERS.ORG Sat Oct 9 12:17:32 2004 From: JKOBLER <@t> PARTNERS.ORG (Kobler, James) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] fibroblast marker(s)? Message-ID: <57531340B9FDD611A8580008026158F1096EF5DE@phsexch26.mgh.harvard.edu> I would appreciate any suggestions for distinguishing fibroblasts from other connective tissue cells using any possible histological techniques. Is it true that there is still not one molecular marker (or combination of markers) that is unambiguously specific for a fibroblast? Thanks very much, Jim Kobler Department of Surgery, Massachusetts General Hospital From lpwenk <@t> sbcglobal.net Sat Oct 9 15:46:18 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] PROFICENCY TRAINING IN HISTOLOGY References: <270614B321ACB44D8C1D91F4F921FDC3627782@NCH01EX02.nch.org> Message-ID: <002f01c4ae41$09c48d80$8cf1ff44@domainnotset.invalid> Competency assessment is done internally by the laboratory, and assesses each person as to their level of competency in terms of knowledge and performing each task. Proficiency assessment is done by an agency outside the laboratory, and assesses the ability of the laboratory as a whole, in its ability to perform specific tasks. There is one agency, CAP, that has set up a proficiency assessment in histology - HistoQIP. A quick overview can be found on the NSH web site at: http://www.nsh.org/education/HistoQip.html Call 800-323-4040 to order. Twice a year, you will get a list of tissue and/or stain requirements. Usually 2 H&E, 2 special stains and 1 IHC. You will recut the tissues from previous cases or control blocks, stain them and submit them. A panel of histotechs and pathologists evaluate your slides/stains for fixation, processing, embedding, microtomy, staining and coverslipping. Your stained slides will be sent back to you along with the panel's assessment of your slides, and a summary of how everyone else who is participating did, so you can compare your lab to other labs. Also included is an explanation of how the stain works, and discussion of troubleshooting the stain. Check with your Cytology or Clinical Pathology departments (Chemistry, Microbiology, etc.). Many of theme may already be participating in their own CAP proficiency assessments. They might still have a catalog. For the CAP catalog, go to: http://www.cap.org On the left side, click on Survey. Then towards the bottom, click on Catalog. The catalog is over 200 pages long, so look in the index for histology. Or, once on the Survey page, click on Request a Catalog. Very educational for labs, in my opinion (no, I'm not on the panel, but our lab does participate). Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Winters, Bert" To: Sent: Friday, October 08, 2004 12:04 PM Subject: [Histonet] PROFICENCY TRAINING IN HISTOLOGY For histotech we have always had a competency check list but have never had a preficiency test per se. Can anyone out there give me a example of what they would include or how to run proficency test for histotechnolgists. Bert Winters HTL (ASCP) NORTHWEST COMMUNITY HOSPITAL ARLINGTON HEIGHTS, ILL EMAIL-----BWINTERS@NCH.ORG ******************* PLEASE NOTE ******************* This E-Mail/telefax message and any documents accompanying this transmission may contain information that is privileged, confidential, and/or exempt from disclosure under applicable law and is intended solely for the addressee(s) named above. If you are not the intended addressee/recipient, you are hereby notified that any use of, disclosure, copying, distribution, or reliance on the contents of this E-Mail/telefax information is strictly prohibited and may result in legal action against you. Please reply to the sender advising of the error in transmission and immediately delete/destroy the message and any accompanying documents. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From je22r <@t> udcf.gla.ac.uk Mon Oct 11 03:01:01 2004 From: je22r <@t> udcf.gla.ac.uk (Julia Edgar) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] bone marrow for immunostaining - how? Message-ID: <002b01c4af68$733e9f80$40e9d182@vet> Dear All I need to collect some bone marrow to test out an antibody to CD45. Can you please tell me how to go about it and how to store the material once collected. Thank you in advance Julia Julia Edgar, BSc (Hons), PhD University of Glasgow Veterinary School Glasgow G61 1QH e-mail: je22r@udcf.gla.ac.uk From susan.wells <@t> bms.com Mon Oct 11 06:31:46 2004 From: susan.wells <@t> bms.com (Susan Q Wells) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] bone marrow for immunostaining - how? In-Reply-To: <002b01c4af68$733e9f80$40e9d182@vet> References: <002b01c4af68$733e9f80$40e9d182@vet> Message-ID: <416A6F22.3010001@bms.com> I'd like to be copied on these responses also. Thanks! Susan Wells Julia Edgar wrote: >Dear All >I need to collect some bone marrow to test out an antibody to CD45. Can you please tell me how to go about it and how to store the material once collected. >Thank you in advance >Julia >Julia Edgar, BSc (Hons), PhD >University of Glasgow Veterinary School >Glasgow G61 1QH >e-mail: je22r@udcf.gla.ac.uk >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From Jackie.O'Connor <@t> abbott.com Mon Oct 11 06:39:46 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] bone marrow for immunostaining - how? Message-ID: Are you collecting human or animal bone marrow? I am able to do CD45 immunostaining on mouse bone marrow by harvesting the entire femur in 10% formalin. The femur is then decalcified in 5% formic acid, and processed for paraffin embedding. Immunostaining is performed routinely, with Citrate buffer antigen retrieval. Jackie O'Connor. "Julia Edgar" Sent by: histonet-bounces@lists.utsouthwestern.edu 10/11/2004 03:01 AM To: cc: Subject: [Histonet] bone marrow for immunostaining - how? Dear All I need to collect some bone marrow to test out an antibody to CD45. Can you please tell me how to go about it and how to store the material once collected. Thank you in advance Julia Julia Edgar, BSc (Hons), PhD University of Glasgow Veterinary School Glasgow G61 1QH e-mail: je22r@udcf.gla.ac.uk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From IKirbis <@t> onko-i.si Mon Oct 11 06:39:00 2004 From: IKirbis <@t> onko-i.si (Kirbis Srebotnik Irena) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] bone marrow for immunostaining - how? Message-ID: you can either suspend bone marrow in tissue culture medium, prepare cytospins (fix in methanol) and stained them by immunocytochemistry or you can perform flow cytometry immunophenotyping on this cell suspension. Irena Srebotnik Kirbis, MSc Institute of Oncology Dept. of Cytopathology Zaloska 2 1000 Ljubljana Slovenia phone +386 1 522 3826 fax +38615879400 -----Original Message----- From: Julia Edgar [mailto:je22r@udcf.gla.ac.uk] Sent: Monday, October 11, 2004 10:01 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] bone marrow for immunostaining - how? Dear All I need to collect some bone marrow to test out an antibody to CD45. Can you please tell me how to go about it and how to store the material once collected. Thank you in advance Julia Julia Edgar, BSc (Hons), PhD University of Glasgow Veterinary School Glasgow G61 1QH e-mail: je22r@udcf.gla.ac.uk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Mon Oct 11 06:41:36 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] bone marrow for immunostaining - how? Message-ID: I see now, Julia would obviously be collecting animal bone marrow. I only harvest "end point" bone marrow. Jackie Susan Q Wells Sent by: histonet-bounces@lists.utsouthwestern.edu 10/11/2004 06:31 AM To: Julia Edgar cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] bone marrow for immunostaining - how? I'd like to be copied on these responses also. Thanks! Susan Wells Julia Edgar wrote: >Dear All >I need to collect some bone marrow to test out an antibody to CD45. Can you please tell me how to go about it and how to store the material once collected. >Thank you in advance >Julia >Julia Edgar, BSc (Hons), PhD >University of Glasgow Veterinary School >Glasgow G61 1QH >e-mail: je22r@udcf.gla.ac.uk >_______________________________________________ >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 From ploykasek <@t> phenopath.com Mon Oct 11 10:31:47 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] immunostaining In-Reply-To: <002101c4acd4$bd5672a0$e1edbcd8@Boswell> Message-ID: Chantel, I second the recommendation about the Dako IHC handbook. It will give you a good overview. Have you checked with your state histo society about any upcoming meetings or seminars? If you do some homework (reading) and go about setting up IHC methodically & carefully, I'm sure you'll do well. Patti Loykasek PhenoPath Laboratories Seattle, WA> If our lab wanted to start learning and doing immunostaining first by hand > then with strainer where would we start. Are there any textbooks out there. > No one in our lab has ever been around immuno's and there is not a lab close > by that we could learn from. Please help!!!! Thank you, Chantel > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From fjones <@t> namsa.com Mon Oct 11 13:09:15 2004 From: fjones <@t> namsa.com (Fawn Jones) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] micronucleus staining Message-ID: <915E55B02E236E4D95258B181EEF6317075D31@namsams01.namsa.int> Does anyone have a wright-giemsa procedure for micronucleus slides that they would be willing to share. Our company wants us to play around with staining times and solutions. Any help would be appreciated. Thanks Fawn Jones From Jackie.O'Connor <@t> abbott.com Mon Oct 11 13:42:58 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Necrosis staining Message-ID: Can anyone give me an idea on a vital stain to differentiate necrotic areas from healthy tissue? Basically, we're looking for a monochromatic stain, easier than an H+E, that we can use for image analysis percentage of necrosis. I thought about toluidine blue, but I'm not sure if it stains dead stuff. Thanks! Happy Monday - Jackie O' From tsanger <@t> biology2.wustl.edu Mon Oct 11 13:51:11 2004 From: tsanger <@t> biology2.wustl.edu (Thomas Sanger) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] calcein bone labeling Message-ID: <8F939618D1D01941B8EC772D5813FEF309BE4F@biology2.wustl.edu> Hello all-??? ? I am trying to get calcein into solution to label growing bones but ??? some particles always seem to remain undissolved.? Is this usual or do I ??? need to adjust the solution slightly?? I have played with the pH but this ??? doesn't seem to fully solve the problem.? Any suggestions would be most appreciated. ??? Thom Sanger From lindas <@t> awesomenet.net Mon Oct 11 14:39:40 2004 From: lindas <@t> awesomenet.net (Linda Davis) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] IHC for Helicobacter Message-ID: <004101c4afca$0e5dcf20$c1a26243@D6JLZ851> Is anyone doing IHC for Helicobacter, and if so what antibody is being used? Thanks. Linda Davis, H.T. (ASCP), B.S. Histology Supervisor Rio Grande Regional Hospital McAllen, TX. From Rcartun <@t> harthosp.org Mon Oct 11 15:02:19 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] IHC for Helicobacter Message-ID: DakoCytomation's polyclonal. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Linda Davis" 10/11/04 03:39PM >>> Is anyone doing IHC for Helicobacter, and if so what antibody is being used? Thanks. Linda Davis, H.T. (ASCP), B.S. Histology Supervisor Rio Grande Regional Hospital McAllen, TX. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Mon Oct 11 15:10:25 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] placental IHC for the novice In-Reply-To: Message-ID: <002401c4afce$587123b0$83020a0a@IHCTech> Julienne, DAKO will provide you with a free handbook on IHC methods. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Julienne Rutherford Sent: Thursday, October 07, 2004 2:48 PM To: Histonet@Lists. Utsouthwestern. Edu Subject: [Histonet] placental IHC for the novice Hello. I'm new to histonet and am not trained in histo or IHC, but am very interested. I'm throwing myself into the fray, and hope you can help me. In response, which I welcome privately or in the forum, please use simple language for I am but a simple woman with a simple dream of studying monkey placentas. I want to study IGF-1 localization in the marmoset monkey placenta. I'm doing a lot of reading regarding methods, but I need help as to what I need to do (and by extension, how to ask for the money to do it in grant applications!). The folks in our histo lab are going to help me and train me but I want to come to them more prepared and I have to provide the specific materials I'll need. I have both 10% NBF-fixed tissues (most sitting around in formalin for a very long time), and I also have frozen, but by this I mean stuck in the regular old lab freezer to freeze conventionally. I will begin snap-freezing placentae we collect from now on, but what can I get out of this old stuff? Can I thaw out what I have and snap-freeze them in OCT? Should I try to do IHC on both the formalin-fixed AND the frozen tissue? In order to build a budget I need to know everything I need to make these slides. So, what kind of slides and coverslips do I need, what are all the chemicals involved, what is a secondary antibody and why do I need it, is a chromagen different from a stain, etc.? Also, is there something akin to and "IHC for Dummies" resource? I've looked at ihcworld.com for protocols but it's over my head at this point. I recognize that this is a skill perfected over many years, but I really would like a primer on the basics so I can have a more informed discussion about it. If the elementary pitch of my questions is inappropriate to the forum, I sincerely apologize. I'm just not sure where else to turn. Please respond to my Texas e-mail if you'd prefer to school (or scold) me privately! Thanks in advance, Julienne Rutherford Research Assistant (Adjunct) Southwest Foundation for Biomedical Research Southwest National Primate Research Center P.O. Box 760549 San Antonio, Texas 78245-0549 phone (210) 258-9864 fax (210) 258-9883 jrutherford@icarus.sfbr.org Ph.D. candidate Indiana University Bloomington, Indiana jnruther@indiana.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From HornHV <@t> archildrens.org Mon Oct 11 15:24:09 2004 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] IHC for Helicobacter Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6322B968@EMAIL.archildrens.org> We get our antibody from CellMarque. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Davis Sent: Monday, October 11, 2004 2:40 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC for Helicobacter Is anyone doing IHC for Helicobacter, and if so what antibody is being used? Thanks. Linda Davis, H.T. (ASCP), B.S. Histology Supervisor Rio Grande Regional Hospital McAllen, TX. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From pjfnefro <@t> duke.edu Mon Oct 11 15:30:35 2004 From: pjfnefro <@t> duke.edu (Pat Flannery) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Calcein Bone Labeling Message-ID: <416AED6B.9070808@duke.edu> Thom- Try dissolving the calcein at 1 mg/ml in 2% Sodium Bicarbonate with 0.9% NaCl. The bicarb makes up for the pH of the calcein. If you want to see the pH difference, try dissolving the calcein in the NaCl first then add the bicarb and you'll see a dramatic color change from dull orange-yellow to brilliant glowing yellow. Hope this helps. -- -Patrick J. (Pat) Flannery Division of Nephrology (that's kidneys to you) Box 3014 (that's NOT "PO" just "Box") Duke University Medical Center Durham, NC 27710 E-mail: pjfnefro@duke.edu (preferred) FLANN002@MC.DUKE.EDU (also works) Voice: (919)660-6863 Fax: (919)684-2929 From pruegg <@t> ihctech.net Mon Oct 11 15:30:57 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] placental IHC for the novice In-Reply-To: Message-ID: <002501c4afd1$36b713d0$83020a0a@IHCTech> I have just been made aware of a book to recommend for an introduction to IHC. It is called: Polak J.M., and Van Noorden S. 1997. INTRODUCTION TO IMMUNOCYTOCHEMISTRY, 2nd edition. Bios Scientific Publishers. ISBN 0-387-91513-3. It costs $46.50 at Amazon.com. The 2nd edition of Jules Elias Immunohistochemistry book can be purchased at ASCP Press, also Amos pointed out an older book by Lawrence True called "Atlas of Diagnostic IHC" which may be found on Amazon, or at least a used copy if it is no longer in print. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Julienne Rutherford Sent: Thursday, October 07, 2004 2:48 PM To: Histonet@Lists. Utsouthwestern. Edu Subject: [Histonet] placental IHC for the novice Hello. I'm new to histonet and am not trained in histo or IHC, but am very interested. I'm throwing myself into the fray, and hope you can help me. In response, which I welcome privately or in the forum, please use simple language for I am but a simple woman with a simple dream of studying monkey placentas. I want to study IGF-1 localization in the marmoset monkey placenta. I'm doing a lot of reading regarding methods, but I need help as to what I need to do (and by extension, how to ask for the money to do it in grant applications!). The folks in our histo lab are going to help me and train me but I want to come to them more prepared and I have to provide the specific materials I'll need. I have both 10% NBF-fixed tissues (most sitting around in formalin for a very long time), and I also have frozen, but by this I mean stuck in the regular old lab freezer to freeze conventionally. I will begin snap-freezing placentae we collect from now on, but what can I get out of this old stuff? Can I thaw out what I have and snap-freeze them in OCT? Should I try to do IHC on both the formalin-fixed AND the frozen tissue? In order to build a budget I need to know everything I need to make these slides. So, what kind of slides and coverslips do I need, what are all the chemicals involved, what is a secondary antibody and why do I need it, is a chromagen different from a stain, etc.? Also, is there something akin to and "IHC for Dummies" resource? I've looked at ihcworld.com for protocols but it's over my head at this point. I recognize that this is a skill perfected over many years, but I really would like a primer on the basics so I can have a more informed discussion about it. If the elementary pitch of my questions is inappropriate to the forum, I sincerely apologize. I'm just not sure where else to turn. Please respond to my Texas e-mail if you'd prefer to school (or scold) me privately! Thanks in advance, Julienne Rutherford Research Assistant (Adjunct) Southwest Foundation for Biomedical Research Southwest National Primate Research Center P.O. Box 760549 San Antonio, Texas 78245-0549 phone (210) 258-9864 fax (210) 258-9883 jrutherford@icarus.sfbr.org Ph.D. candidate Indiana University Bloomington, Indiana jnruther@indiana.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kathy.Paton <@t> waitematadhb.govt.nz Mon Oct 11 15:48:38 2004 From: Kathy.Paton <@t> waitematadhb.govt.nz (Kathy Paton (WDHB)) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] IHC for Helicobacter Message-ID: Dakocytomation H.py is good Kathy Paton -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Davis Sent: Tuesday, 12 October 2004 08:40 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC for Helicobacter Is anyone doing IHC for Helicobacter, and if so what antibody is being used? Thanks. Linda Davis, H.T. (ASCP), B.S. Histology Supervisor Rio Grande Regional Hospital McAllen, TX. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From NBerghoff <@t> cvm.tamu.edu Mon Oct 11 16:01:11 2004 From: NBerghoff <@t> cvm.tamu.edu (Nora Berghoff) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Movat's stain/hematoxylin Message-ID: Hello everyone! I am new to this list, so first I would like to say hi to everyone!! I have been reading the archives and find that they are a wonderful source of knowledge! I am also new in the world of histology - which is why I am asking for your help: I want to stain canine intestinal biopsies for collagen using Russell's modification of Movat's pentachrome stain. I have stained my first sections today and have encountered some problems: First, most of my muscle tissue is not really red, but rather orange/yellowish from the saffron, as it seems. But some areas of muscle have stained red. Fibrin does seem to be stained well (if what I have identified as fibrin is in fact fibrin...;)) What can cause such irregular staining? How long after cutting of the sections are they "stable" (meaning they don't lose any staining ability) on the slide? Could it be that my cut sections are old and "dried out"? I have had a histo lab embed and cut them, which is about five weeks ago...so I figured something may have happened to the sections in the meantime? Also, my elastic fibers and nuclei are not really black (I stained them according to the protocol for 15 minutes in hematoxylin sol.). I found it difficult to find the right hematoxylin (there are so many of them!), so maybe I have the wrong one after all... Does anyone know how long the individual staining solutions are stable after preparing them? Maybe my solutions are bad... So many questions and "maybe's" - I would really appreciate it very much if I could get some suggestions on any of them! Thank you so much in advance!! Nora Berghoff Research Assistant Gastrointestinal Laboratory Dept of Small Animal Clinical Sciences College of Veterinary Medicine Texas A&M University From gcallis <@t> montana.edu Mon Oct 11 17:30:59 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] DAKO Handbook on DAKO website In-Reply-To: <002401c4afce$587123b0$83020a0a@IHCTech> References: <002401c4afce$587123b0$83020a0a@IHCTech> Message-ID: <6.0.0.22.1.20041011162938.01b2e378@gemini.msu.montana.edu> You can download the Handbook from DAKOCYTOMATION website Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From weneng2004 <@t> yahoo.com Mon Oct 11 17:45:34 2004 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] cut the old paraffin block Message-ID: <20041011224534.12457.qmail@web53409.mail.yahoo.com> Dear histonetters, I was trying to cut the about 3 years old paraffin today but failed. I keep all my paraffin blocks in file drawer at room temperature. I tried to cut the sample after it was put on ice water for 1hr, 3 hr and even 5 hrs but still feel dry. Anybody know why I cannot make it? Any suggestion would be highly appreciated! Thanks, Wendy --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From lindas <@t> awesomenet.net Mon Oct 11 18:03:58 2004 From: lindas <@t> awesomenet.net (Linda Davis) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] IHC for Helicobacter Message-ID: <002c01c4afe6$988b9580$d0a16243@D6JLZ851> Thank all of you who responsed to my question about IHC for Helicobacter. Linda Davis From pruegg <@t> ihctech.net Mon Oct 11 18:15:01 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] cut the old paraffin block In-Reply-To: <20041011224534.12457.qmail@web53409.mail.yahoo.com> Message-ID: <002e01c4afe8$221ce910$83020a0a@IHCTech> First try reembedding it. Sometimes I put glycerine in the ice bath that helps soften brittle tissue as well. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of wen eng Sent: Monday, October 11, 2004 3:46 PM To: histonet Subject: [Histonet] cut the old paraffin block Dear histonetters, I was trying to cut the about 3 years old paraffin today but failed. I keep all my paraffin blocks in file drawer at room temperature. I tried to cut the sample after it was put on ice water for 1hr, 3 hr and even 5 hrs but still feel dry. Anybody know why I cannot make it? Any suggestion would be highly appreciated! Thanks, Wendy --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From S2118495 <@t> student.rmit.edu.au Mon Oct 11 18:21:58 2004 From: S2118495 <@t> student.rmit.edu.au (Christopher Ian Hill) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Artifacts or real - unusual mouse liver apperance in H&E Message-ID: <1097536918.912a9f60S2118495@student.rmit.edu.au> Can anyone with experiance processing mouse liver help? We have consistently been getting cytoplasmic clearing and apparent autophagy of hepatocytes in mouse liver sections, as well as some regions which appear to be showing localised necrosis some with and some without infiltrate. We have tried different strains of mice and different people doing all steps. We have even tried different fixatives (Bouins and 10% NBF). These mice have not been treated with anything, and we see this both immediatly after they arrive as well as after a week and after months. Is there something different about processing mouse livers that we should know? Or may we be seeign the manifestation of a hepatitis? Chris Hill From pruegg <@t> ihctech.net Mon Oct 11 18:23:32 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] knife holder for microtome in cryostat Message-ID: <003101c4afe9$5342bd20$83020a0a@IHCTech> Folks, I have a Reichert-Jung Cryocut 1800 cryostat, the microtome inside says Leica 2020, I am looking for a stage to fit on the microtome to hold a steel knife, the one on it now is for disposable blades. I set up the Intermedics Tape Transfer system in this cryostat and want to use a tungsten carbide D profile knife to cut calcified bone. Perhaps someone has one of these stages for this microtome they could sell me? Patsy Ruegg pruegg@ihctech.net From JQB7 <@t> CDC.GOV Mon Oct 11 18:35:26 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] cut the old paraffin block Message-ID: I also would re-embed it. Then I would put it in a container of crushed ice to which you have added a little ammonia. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of wen eng Sent: Mon 10/11/2004 6:45 PM To: histonet Cc: Subject: [Histonet] cut the old paraffin block Dear histonetters, I was trying to cut the about 3 years old paraffin today but failed. I keep all my paraffin blocks in file drawer at room temperature. I tried to cut the sample after it was put on ice water for 1hr, 3 hr and even 5 hrs but still feel dry. Anybody know why I cannot make it? Any suggestion would be highly appreciated! Thanks, Wendy --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kwuny <@t> EMAIL.CS.NSW.GOV.AU Mon Oct 11 20:23:38 2004 From: kwuny <@t> EMAIL.CS.NSW.GOV.AU (Young Kwun) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Collagen IV antibody In-Reply-To: <6.1.0.6.2.20041007153416.02479828@tigger.uic.edu> Message-ID: <20041012111946.SM05496@crgcsls814> Lu, You can get anti-Collagen IV, alpha 5 chain (Clone A7, Cat No: MC-885) from KAMIYA BIOMEDICAL COMPANY, Seatle, WA. www.kamiyabiomedical.com They also have alpha 1 and alpha 3 chains as well. Young Young Kwun Senior Hospital Scientist Dept of Anatomical Pathology Concord Hospital Concord NSW 2139 Australia Phone:61-2-9767 6075 Fax:61-2-9767 8427 Email:kwuny@email.cs.nsw.gov.au -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lu Leach Sent: Friday, 8 October 2004 6:37 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Collagen IV antibody Dear Histonetters, Can anyone tell me where to acquire Collagen IV A5 antibody? Thanks, Lu Leach University of Illinois at Chicago Research Pathology Core 312-996-3869 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "This message is intended for the addressee named and may contain confidential information. If you are not the intended recipient, please destroy it and notify the sender. Views expressed in this message are those of the individual sender, and are not necessarily the views of the Central Sydney Area Health Service." From pengbw <@t> sjtu.edu.cn Mon Oct 11 20:27:37 2004 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Smears Message-ID: <20041012012737.376C61108B92@sjtu.edu.cn> Thank you all who respond to help solve my problem. I use 1640 to stand for cell culture medium RPMI-1640, sorry for the confusing. Yes, I prepared direct smear from cell suspension. Irena\'s suggestion to reducing fluid when preparing smears indeed can ease the problem. Baowei Peng Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China From Inga.Hansson <@t> neuro.uu.se Tue Oct 12 04:41:39 2004 From: Inga.Hansson <@t> neuro.uu.se (Inga Hansson) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] anti-GFP Message-ID: Does anyone know if the mixed monoclonal anti-GFP from Roche works on cryosections (or paraffin sections ) and in that case what fixation to use? Thanks in advance! Inga -- Inga Hansson dept. neuroscience, div. neurobiology PO Box 587 Biomedical Centre Husargatan 3 S-751 23 Uppsala SWEDEN phone:+46-18-4714384 fax: +46-18-559017 From jluis.palazon <@t> icman.csic.es Tue Oct 12 05:33:37 2004 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] bone decalcification Message-ID: <20041012103337.5060C3326BD@perceval.uca.es> Dear List-members I have to dacalcify some fish vertebrae in order to see the vertebral articulation and intervertebral discs, and have to use a quieck method. I usually decalcify with EDTA but this method is very slow. I would appreciate your advise. Thanks in advance! Jos? Luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es From bruyntjes <@t> voeding.tno.nl Tue Oct 12 06:49:43 2004 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Eyes Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079D2F@ntexch1.voeding.tno.nl> Hi all Is anyone of you familiar with a staining of (chicken) eyes which visualizes the bowman's membrane? J.P. Bruijntjes TNO Nutrition and Food Research Toxicology and Appllied Pharmacology Utrechtse weg 48 PO Box 360 3700 AJ Zeist The Netherlands T +31 30 6944480 F +31 30 6960264 This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From akbitting <@t> geisinger.edu Tue Oct 12 06:40:01 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] H. pylori Message-ID: We're perplexed. We've been using Dako's H. pylori AB with Proteinase K antigen retreival for years now and have never had this problem. Our control is teeming with organisms but some are staining brown and others are more of a bluish color. I'm sure they are all H. Pyl organisms, but I can't figure out why they aren't all staining the same. Dako says they haven't had any complaints about their Lot number. We've tried different vials from that same Lot # and both act the same. Trying different dilutions didn't help. Now, we're extending the Proteinase K time. After this, we're out of ideas. Help! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From kmerriam2003 <@t> yahoo.com Tue Oct 12 09:44:17 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] alternates to OCT embedding media Message-ID: <20041012144417.57647.qmail@web52506.mail.yahoo.com> Hello all, I was wondering if anyone knew of alternates to OCT-brand cryo-embedding media. I know that there are other brands of the stuff, but it is my understanding that they are all basically made out of the same thing. Does anyone know of any other media that is used to prepare frozen blocks? Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Express yourself with Y! Messenger! Free. Download now. From Don.Birgerson <@t> leica-microsystems.com Tue Oct 12 09:56:43 2004 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] knife holder for microtome in cryostat Message-ID: Hi Patsy, While the 1800 Cryostat is one generation behind the CM1850, the current CN holder (steel/carbide ) will fit your 2020 Microtome. Give me a call for numbers as this will depend on your current holder. Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Patsy Ruegg" To: Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] knife holder for microtome in cryostat western.edu 10/11/2004 06:23 PM Folks, I have a Reichert-Jung Cryocut 1800 cryostat, the microtome inside says Leica 2020, I am looking for a stage to fit on the microtome to hold a steel knife, the one on it now is for disposable blades. I set up the Intermedics Tape Transfer system in this cryostat and want to use a tungsten carbide D profile knife to cut calcified bone. Perhaps someone has one of these stages for this microtome they could sell me? Patsy Ruegg pruegg@ihctech.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From vazquezr <@t> ohsu.edu Tue Oct 12 09:58:09 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] alternates to OCT embedding media Message-ID: TBS makes a embedding media, but OCT is better. If you are like me and constantly using the media during surgery, TBS bottle is much harder to squeeze, as in comparison to the OCT bottle. And the quality of OCT is better. Robyn OHSU Portland,OR >>> Kim Merriam 10/12/2004 7:44:17 AM >>> Hello all, I was wondering if anyone knew of alternates to OCT-brand cryo-embedding media. I know that there are other brands of the stuff, but it is my understanding that they are all basically made out of the same thing. Does anyone know of any other media that is used to prepare frozen blocks? Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Express yourself with Y! Messenger! Free. Download now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Tue Oct 12 10:00:29 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] alternates to OCT embedding media - tangent from Message-ID: One of the ways American is different from English is in the word alternate, which in American, is so often used as an alternative to alternative. In English this is never so, keeping the meaning of something on the lines or "you-me-you-me" etc. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kim Merriam [mailto:kmerriam2003@yahoo.com] Sent: 12 October 2004 15:44 To: Histonet Subject: [Histonet] alternates to OCT embedding media Hello all, I was wondering if anyone knew of alternates to OCT-brand cryo-embedding media. I know that there are other brands of the stuff, but it is my understanding that they are all basically made out of the same thing. Does anyone know of any other media that is used to prepare frozen blocks? Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Express yourself with Y! Messenger! Free. Download now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Tue Oct 12 10:03:42 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] alternates to OCT embedding media Message-ID: What is this stuff made from? It always reminds me of wallpaper hanging gel - something like "Polycell" in England. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Robyn Vazquez [mailto:vazquezr@ohsu.edu] Sent: 12 October 2004 15:58 To: histonet@lists.utsouthwestern.edu; kmerriam2003@yahoo.com Subject: Re: [Histonet] alternates to OCT embedding media TBS makes a embedding media, but OCT is better. If you are like me and constantly using the media during surgery, TBS bottle is much harder to squeeze, as in comparison to the OCT bottle. And the quality of OCT is better. Robyn OHSU Portland,OR >>> Kim Merriam 10/12/2004 7:44:17 AM >>> Hello all, I was wondering if anyone knew of alternates to OCT-brand cryo-embedding media. I know that there are other brands of the stuff, but it is my understanding that they are all basically made out of the same thing. Does anyone know of any other media that is used to prepare frozen blocks? Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Express yourself with Y! Messenger! Free. Download now. _______________________________________________ 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 From gentras <@t> vetmed.auburn.edu Tue Oct 12 10:27:54 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:24:09 2005 Subject: Fwd: [Histonet] alternates to OCT embedding media Message-ID: <6.0.1.1.0.20041012102337.01b05bc0@mailhost.vetmed.auburn.edu> Hello, I'm currently using Tissue Freezing Medium (TFM), TBS. Itt is manufactured by Triangle Biomedical sciences and can be purchased through either Fisher Scientific or VWR. The VWR catalog number is 15148-031. It works fine for me. Best wishes. Atoska >Date: Tue, 12 Oct 2004 07:44:17 -0700 (PDT) >From: Kim Merriam >To: Histonet >X-Scan-Signature: fafe997e7c8aede70e4d53d18f8e9e53 >X-Content-Filtered-By: Mailman/MimeDel 2.1.3 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.3 >List-Id: For the exchange of information pertaining to histotechnology and > related fields >List-Unsubscribe: >, > > >List-Archive: >List-Post: >List-Help: >List-Subscribe: , > >Sender: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: 1d06f19e0b5ef274560ff195311cd0ef >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >Subject: [Histonet] alternates to OCT embedding media >X-Spam-Checker-Version: SpamAssassin 2.63 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: *** >X-Spam-Status: No, hits=3.2 required=6.5 tests=FROM_ENDS_IN_NUMS,RCVD_IN_DSBL, > RCVD_IN_NJABL,RCVD_IN_NJABL_PROXY autolearn=no version=2.63 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >X-spam: ESP<37>=RBL:<0> RDNS:<0> SHA:<34> UHA:<0> SLS:<0> BAYES:<3> >SPF:<0> Porn Dictionary (TRU5):<0> Spam Dictionary (TRU5):<0> HTML >Dictionary (TRU5):<0> DRUGS2:<0> Obscenities Dictionary (TRU5):<0> >CAN-SPAM Compliance Dictionary (TRU5):<0> NigeriaScam Dictionary >(TRU5):<0> Embed HTML Dictionary (TRU5):<0> URL Dictionary (TRU5):<0> >porn2:<0> > >Hello all, > >I was wondering if anyone knew of alternates to OCT-brand cryo-embedding >media. I know that there are other brands of the stuff, but it is my >understanding that they are all basically made out of the same >thing. Does anyone know of any other media that is used to prepare frozen >blocks? > > > >Kim Merriam >Novartis >Cambridge, MA > >--------------------------------- >Do you Yahoo!? >Express yourself with Y! Messenger! Free. Download now. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gentras <@t> vetmed.auburn.edu Tue Oct 12 10:29:46 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:24:09 2005 Subject: Fwd: Re: [Histonet] alternates to OCT embedding media Message-ID: <6.0.1.1.0.20041012102904.01b63420@mailhost.vetmed.auburn.edu> Hello, please who is your source for OCT these days??? Thanks, Atoska >X-Server-Uuid: E8EE9E4E-CE80-4A3D-84F1-1599A05BA687 >X-Mailer: Novell GroupWise Internet Agent 6.5.2 Beta >Date: Tue, 12 Oct 2004 07:58:09 -0700 >From: "Robyn Vazquez" >To: histonet@lists.utsouthwestern.edu, kmerriam2003@yahoo.com >X-WSS-ID: 6D752E8E2PW11625-01-01 >X-Scan-Signature: 21b68cbd38606e624bfc9734c34a1e71 >X-Content-Filtered-By: Mailman/MimeDel 2.1.3 >Cc: >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.3 >List-Id: For the exchange of information pertaining to histotechnology and > related fields >List-Unsubscribe: >, > > >List-Archive: >List-Post: >List-Help: >List-Subscribe: , > >Sender: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: 2fb82c3b59105628058b6e8043468fa4 >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >Subject: Re: [Histonet] alternates to OCT embedding media >X-Spam-Checker-Version: SpamAssassin 2.63 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: * >X-Spam-Status: No, hits=1.1 required=6.5 tests=MAILTO_TO_SPAM_ADDR > autolearn=no version=2.63 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >X-spam: ESP<36>=RBL:<0> RDNS:<0> SHA:<33> UHA:<0> SLS:<0> BAYES:<3> >SPF:<0> Porn Dictionary (TRU5):<0> Spam Dictionary (TRU5):<0> HTML >Dictionary (TRU5):<0> DRUGS2:<0> Obscenities Dictionary (TRU5):<0> >CAN-SPAM Compliance Dictionary (TRU5):<0> NigeriaScam Dictionary >(TRU5):<0> Embed HTML Dictionary (TRU5):<0> URL Dictionary (TRU5):<0> >porn2:<0> > >TBS makes a embedding media, but OCT is better. If you are like me and >constantly using the media during surgery, TBS bottle is much harder to >squeeze, as in comparison to the OCT bottle. And the quality of OCT is >better. > >Robyn >OHSU >Portland,OR > > >>> Kim Merriam 10/12/2004 7:44:17 AM >>> > >Hello all, > >I was wondering if anyone knew of alternates to OCT-brand >cryo-embedding media. I know that there are other brands of the stuff, >but it is my understanding that they are all basically made out of the >same thing. Does anyone know of any other media that is used to prepare >frozen blocks? > > > >Kim Merriam >Novartis >Cambridge, MA > >--------------------------------- >Do you Yahoo!? >Express yourself with Y! Messenger! Free. Download now. >_______________________________________________ >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 From gcallis <@t> montana.edu Tue Oct 12 10:49:32 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Alternates to OCT embedding media In-Reply-To: <20041012144417.57647.qmail@web52506.mail.yahoo.com> References: <20041012144417.57647.qmail@web52506.mail.yahoo.com> Message-ID: <6.0.0.22.1.20041012090150.01b22198@gemini.msu.montana.edu> Kim, Caveat: Be very aware that they are NOT the same. I strongly suggest try samples of different brands. Vendors (bless their hearts) have always provided us with samples. Our experience (a huge eye opener) with a new cryoembedding product compared in parallel to OCT was a huge eyeopener. We always fill mouse intestinal lumen with cryoembedding media, also mouse lung. The new product failed to work as villi were not supported, and sections were compressed and torn up. When I asked vendor if their new cryomedia had ever been tested on murine, the answer was "NO". They had only tried it for human application. I appreciated their honesty and they also learned something too. This is no way meant their product was defective, it just didn't work for our research application and not compatible for our funky mouse work. So I strongly advise anyone to test cryoembedding media with their applications, sectioning temperature requirements, failure to stay around tissue during sectioning, etc, etc, to may sure you have a media optimal for your work. We have found some seem watery, have poor sectioning or support qualities. These products ONLY failed for us and others may not experience the same problems. It is strictly our laboratory's choice to stay with OCT, but we continue to try any new cryomedias available with feedback (privately) to the vendor(s). Have fun experimenting - it takes time but the effort pays off. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman Bozeman MT 59717-3610 At 08:44 AM 10/12/2004, you wrote: >Hello all, > >I was wondering if anyone knew of alternates to OCT-brand cryo-embedding >media. I know that there are other brands of the stuff, but it is my >understanding that they are all basically made out of the same >thing. Does anyone know of any other media that is used to prepare frozen >blocks? > > > >Kim Merriam >Novartis >Cambridge, MA > >--------------------------------- >Do you Yahoo!? >Express yourself with Y! Messenger! Free. Download now. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From min_minlu <@t> hotmail.com Tue Oct 12 10:48:22 2004 From: min_minlu <@t> hotmail.com (Min-Min Lu) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Automated coversliper Message-ID: Hi all, I am looking for a small size Automated coverslipper for our Histology Core Lab. We only have about 100 HE slides to be stained and covered each week. We do it all manually. Is it a good idea to have an Automated coverslipper? Which company makes good Automated coverslipper? Is it reliable and easy to operate? Your information is greatly appreciated. MinMin Lu Director, Histology and Gene Expression Core Molecular Cardiology Research Center University of Pennsylvania 982 BRB II 421 Curie Blvd Philadelphia, PA 19104 215-898-0251 _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From gcallis <@t> montana.edu Tue Oct 12 10:56:29 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] What OCT contains In-Reply-To: References: Message-ID: <6.0.0.22.1.20041012095049.01b56c98@gemini.msu.montana.edu> The ingredients of OCT are on the bottle, a Polyvinyl alcohol, polyethylene glyol and inert ingredients. They give percentages but not molecular weights, that is proprietary. I had an interesting discussion with Dr. McCormick, inventor of OCT for Miles, many years ago and he indicated he could change the ingredients and probably the concentrations to have a cryomedia specific for other temperatures, colder or warmer than the temperature range for OCT. Fisherbrand cryoembedding media ingredients used to be on the bottle too. At 09:03 AM 10/12/2004, you wrote: >What is this stuff made from? >It always reminds me of wallpaper hanging gel - something like "Polycell" >in England. > >Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path > Consultant Pathologist > Rotherham General Hospital > South Yorkshire > England > terry.marshall@rothgen.nhs.uk > >-----Original Message----- >From: Robyn Vazquez [mailto:vazquezr@ohsu.edu] >Sent: 12 October 2004 15:58 >To: histonet@lists.utsouthwestern.edu; kmerriam2003@yahoo.com >Subject: Re: [Histonet] alternates to OCT embedding media > > >TBS makes a embedding media, but OCT is better. If you are like me and >constantly using the media during surgery, TBS bottle is much harder to >squeeze, as in comparison to the OCT bottle. And the quality of OCT is >better. > >Robyn >OHSU >Portland,OR > > >>> Kim Merriam 10/12/2004 7:44:17 AM >>> > >Hello all, > >I was wondering if anyone knew of alternates to OCT-brand >cryo-embedding media. I know that there are other brands of the stuff, >but it is my understanding that they are all basically made out of the >same thing. Does anyone know of any other media that is used to prepare >frozen blocks? > > > >Kim Merriam >Novartis >Cambridge, MA > >--------------------------------- >Do you Yahoo!? >Express yourself with Y! Messenger! Free. Download now. >_______________________________________________ >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 Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From jkiernan <@t> uwo.ca Tue Oct 12 11:00:05 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Movat's stain/hematoxylin References: Message-ID: <416BFF85.1B24D6C@uwo.ca> Welcome to Histonet! You have several questions. Try again, asking just one with each email enquiry. If you are "new in the world of histology" you have been thrown in at the deep end with Movat's pentachrome (even deeper with someone's modification). Does your boss know anything about these methods? There are plenty of simple (3 or 4 steps) ways to stain collagen. You don't need a method such as Movat's pentachrome (10 solutions and 14 steps) to do such a simple stainng job. You should persuade your boss to buy the lab one or more books about staining methods. Books in this field cost about $100. How many slides stained by the Movat method would you get from a commercial lab for $100? Ten? Five? (Will someone tell us?) The van Gieson stain shows collagen very well using only two staining solutions. The submucosa of the intestine always stands out very well. If you need to see very thin fibres, the related picro-sirius red method is just as easy to do. For multiple colors (if that's a necessity) one of the one-step trichrome techniques will probably be adequate. All the methods mentioned in this paragraph are suitable for a beginner and can be completed in 15-20 minutes. John Kiernan London, Canada. _______________________________________________ Nora Berghoff wrote: > > Hello everyone! > > I am new to this list, so first I would like to say hi to everyone!! I > have been reading the archives and find that they are a wonderful source > of knowledge! > > I am also new in the world of histology - which is why I am asking for > your help: > I want to stain canine intestinal biopsies for collagen using Russell's > modification of Movat's pentachrome stain. I have stained my first > sections today and have encountered some problems: > > First, most of my muscle tissue is not really red, but rather > orange/yellowish from the saffron, as it seems. But some areas of muscle > have stained red. > Fibrin does seem to be stained well (if what I have identified as > fibrin is in fact fibrin...;)) > > What can cause such irregular staining? How long after cutting of the > sections are they "stable" (meaning they don't lose any staining > ability) on the slide? > Could it be that my cut sections are old and "dried out"? I have had a > histo lab embed and cut them, which is about five weeks ago...so I > figured something may have happened to the sections in the meantime? > > Also, my elastic fibers and nuclei are not really black (I stained them > according to the protocol for 15 minutes in hematoxylin sol.). I found > it difficult to find the right hematoxylin (there are so many of them!), > so maybe I have the wrong one after all... > > Does anyone know how long the individual staining solutions are stable > after preparing them? Maybe my solutions are bad... > > So many questions and "maybe's" - I would really appreciate it very > much if I could get some suggestions on any of them! > > Thank you so much in advance!! > > Nora Berghoff > > Research Assistant > Gastrointestinal Laboratory > Dept of Small Animal Clinical Sciences > College of Veterinary Medicine > Texas A&M University > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Tue Oct 12 10:48:35 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:09 2005 Subject: Fwd: Re: [Histonet] alternates to OCT embedding media Message-ID: Atoska, I get it from Cardinal Health Medical Products and Services, they used to be Allegiance. They do not have an catalog to send. They are online only now. I have used both and you can sure tell the difference. Robyn OHSU Portland, OR >>> "Atoska S. Gentry" 10/12/2004 8:29:46 AM >>> Hello, please who is your source for OCT these days??? Thanks, Atoska >X-Server-Uuid: E8EE9E4E-CE80-4A3D-84F1-1599A05BA687 >X-Mailer: Novell GroupWise Internet Agent 6.5.2 Beta >Date: Tue, 12 Oct 2004 07:58:09 -0700 >From: "Robyn Vazquez" >To: histonet@lists.utsouthwestern.edu, kmerriam2003@yahoo.com >X-WSS-ID: 6D752E8E2PW11625-01-01 >X-Scan-Signature: 21b68cbd38606e624bfc9734c34a1e71 >X-Content-Filtered-By: Mailman/MimeDel 2.1.3 >Cc: >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.3 >List-Id: For the exchange of information pertaining to histotechnology and > related fields >List-Unsubscribe: >, > > >List-Archive: >List-Post: >List-Help: >List-Subscribe: , > >Sender: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: 2fb82c3b59105628058b6e8043468fa4 >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >Subject: Re: [Histonet] alternates to OCT embedding media >X-Spam-Checker-Version: SpamAssassin 2.63 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: * >X-Spam-Status: No, hits=1.1 required=6.5 tests=MAILTO_TO_SPAM_ADDR > autolearn=no version=2.63 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >X-spam: ESP<36>=RBL:<0> RDNS:<0> SHA:<33> UHA:<0> SLS:<0> BAYES:<3> >SPF:<0> Porn Dictionary (TRU5):<0> Spam Dictionary (TRU5):<0> HTML >Dictionary (TRU5):<0> DRUGS2:<0> Obscenities Dictionary (TRU5):<0> >CAN-SPAM Compliance Dictionary (TRU5):<0> NigeriaScam Dictionary >(TRU5):<0> Embed HTML Dictionary (TRU5):<0> URL Dictionary (TRU5):<0> >porn2:<0> > >TBS makes a embedding media, but OCT is better. If you are like me and >constantly using the media during surgery, TBS bottle is much harder to >squeeze, as in comparison to the OCT bottle. And the quality of OCT is >better. > >Robyn >OHSU >Portland,OR > > >>> Kim Merriam 10/12/2004 7:44:17 AM >>> > >Hello all, > >I was wondering if anyone knew of alternates to OCT-brand >cryo-embedding media. I know that there are other brands of the stuff, >but it is my understanding that they are all basically made out of the >same thing. Does anyone know of any other media that is used to prepare >frozen blocks? > > > >Kim Merriam >Novartis >Cambridge, MA > >--------------------------------- >Do you Yahoo!? >Express yourself with Y! Messenger! Free. Download now. >_______________________________________________ >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 From amosbrooks <@t> earthlink.net Tue Oct 12 11:15:27 2004 From: amosbrooks <@t> earthlink.net (amosbrooks@earthlink.net) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] DAKO Handbook on DAKO website Message-ID: <6143096.1097597729290.JavaMail.root@bert.psp.pas.earthlink.net> Gayle, When I first suggested the handbook I was going to include a link to it but when I went to the DAKO website to find it I found they had changed the site a lot since I was last there and I couldn't find the hand book. Do you happen to have a URL for it? I knew I should have bookmarked it when I saw it last! thanks, Amos Message: 12 Date: Mon, 11 Oct 2004 16:30:59 -0600 From: Gayle Callis Subject: [Histonet] DAKO Handbook on DAKO website To: Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20041011162938.01b2e378@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed You can download the Handbook from DAKOCYTOMATION website Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From juan.gutierrez <@t> christushealth.org Tue Oct 12 11:15:22 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] alternates to OCT embedding media Message-ID: I believe they're called Cardinal these days, use to be Baxter and Allegiance for a while. The phone number I have is for Baxter, but I think it still works. 1-800-964-5227 -----Original Message----- From: Atoska S. Gentry [mailto:gentras@vetmed.auburn.edu] Sent: Tuesday, October 12, 2004 10:30 AM To: Histonet Subject: Fwd: Re: [Histonet] alternates to OCT embedding media Hello, please who is your source for OCT these days??? Thanks, Atoska >X-Server-Uuid: E8EE9E4E-CE80-4A3D-84F1-1599A05BA687 >X-Mailer: Novell GroupWise Internet Agent 6.5.2 Beta >Date: Tue, 12 Oct 2004 07:58:09 -0700 >From: "Robyn Vazquez" >To: histonet@lists.utsouthwestern.edu, kmerriam2003@yahoo.com >X-WSS-ID: 6D752E8E2PW11625-01-01 >X-Scan-Signature: 21b68cbd38606e624bfc9734c34a1e71 >X-Content-Filtered-By: Mailman/MimeDel 2.1.3 >Cc: >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.3 >List-Id: For the exchange of information pertaining to histotechnology and > related fields >List-Unsubscribe: >, > > >List-Archive: >List-Post: >List-Help: >List-Subscribe: , > >Sender: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: 2fb82c3b59105628058b6e8043468fa4 >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >Subject: Re: [Histonet] alternates to OCT embedding media >X-Spam-Checker-Version: SpamAssassin 2.63 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: * >X-Spam-Status: No, hits=1.1 required=6.5 tests=MAILTO_TO_SPAM_ADDR > autolearn=no version=2.63 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >X-spam: ESP<36>=RBL:<0> RDNS:<0> SHA:<33> UHA:<0> SLS:<0> BAYES:<3> >SPF:<0> Porn Dictionary (TRU5):<0> Spam Dictionary (TRU5):<0> HTML >Dictionary (TRU5):<0> DRUGS2:<0> Obscenities Dictionary (TRU5):<0> >CAN-SPAM Compliance Dictionary (TRU5):<0> NigeriaScam Dictionary >(TRU5):<0> Embed HTML Dictionary (TRU5):<0> URL Dictionary (TRU5):<0> >porn2:<0> > >TBS makes a embedding media, but OCT is better. If you are like me and >constantly using the media during surgery, TBS bottle is much harder to >squeeze, as in comparison to the OCT bottle. And the quality of OCT is >better. > >Robyn >OHSU >Portland,OR > > >>> Kim Merriam 10/12/2004 7:44:17 AM >>> > >Hello all, > >I was wondering if anyone knew of alternates to OCT-brand >cryo-embedding media. I know that there are other brands of the stuff, >but it is my understanding that they are all basically made out of the >same thing. Does anyone know of any other media that is used to prepare >frozen blocks? > > > >Kim Merriam >Novartis >Cambridge, MA > >--------------------------------- >Do you Yahoo!? >Express yourself with Y! Messenger! Free. Download now. >_______________________________________________ >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 From nancy.troiano <@t> yale.edu Tue Oct 12 11:18:54 2004 From: nancy.troiano <@t> yale.edu (Nancy W. Troiano) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] calcein labeling of bone Message-ID: <5.2.1.1.2.20041012121336.01c52ea8@email.med.yale.edu> We prepare our calcein for mouse injections as follows: Dissolve 0.219 grams NaCl in 25 ml distilled water. Add 0.500 grams sodium bicarbonate and stir until dissolved. Add 0.250g calcein powder slowly while gently heating above solution (be careful - it can foam up). Stir until all calcein dissolves. Filter sterilize. Mouse dose of calcein is 30 mg/kg. Nancy Troiano Associate in Research III Yale Core Center for Musculoskeletal Disorders PO Box 208071 New Haven, CT 06520 (203)785-5136 From vazquezr <@t> ohsu.edu Tue Oct 12 10:52:05 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] alternates to OCT embedding media Message-ID: Terry, I believe it is unscented hairgel. I would imagine that it is less toxic and less sticky:) Robyn OHSU Portland, OR >>> "Marshall Terry Dr, Consultant Histopathologist" 10/12/2004 8:03:42 AM >>> What is this stuff made from? It always reminds me of wallpaper hanging gel - something like "Polycell" in England. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Robyn Vazquez [mailto:vazquezr@ohsu.edu] Sent: 12 October 2004 15:58 To: histonet@lists.utsouthwestern.edu; kmerriam2003@yahoo.com Subject: Re: [Histonet] alternates to OCT embedding media TBS makes a embedding media, but OCT is better. If you are like me and constantly using the media during surgery, TBS bottle is much harder to squeeze, as in comparison to the OCT bottle. And the quality of OCT is better. Robyn OHSU Portland,OR >>> Kim Merriam 10/12/2004 7:44:17 AM >>> Hello all, I was wondering if anyone knew of alternates to OCT-brand cryo-embedding media. I know that there are other brands of the stuff, but it is my understanding that they are all basically made out of the same thing. Does anyone know of any other media that is used to prepare frozen blocks? Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Express yourself with Y! Messenger! Free. Download now. _______________________________________________ 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 From MElliott <@t> mrl.ubc.ca Tue Oct 12 11:37:01 2004 From: MElliott <@t> mrl.ubc.ca (Mark Elliott) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] alternates to OCT embedding media Message-ID: Kim We use Cryomatrix which comes from Shandon (we order it through Fisher). What is nice about it is you can get it in a gallon jug, which is good for us because we use it quite a bit (inflate human lung samples with it) and is a lot chepaer then OCT. If you have empty OCT bottles you can just refill with Cryomatrix. I think it may come in a smaller bottle (OCT size). It also used to come in different colours if you wanted to colour code things. The dye didn't affect any of the staining we did-washed out in processing of the slides. W. Mark Elliott, PhD Research Associate James Hogg iCAPTURE Centre for Cardiovascular and Pulmonary Research Room 166, St. Paul's Hospital 1081 Burrard Street Vancouver, BC CAnada V6Z 1Y6 >>> Kim Merriam 10/12/2004 7:44:17 AM >>> Hello all, I was wondering if anyone knew of alternates to OCT-brand cryo-embedding media. I know that there are other brands of the stuff, but it is my understanding that they are all basically made out of the same thing. Does anyone know of any other media that is used to prepare frozen blocks? Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Express yourself with Y! Messenger! Free. Download now. From contact <@t> excaliburpathology.com Tue Oct 12 11:44:08 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] cut the old paraffin blocks Message-ID: <20041012164408.55323.qmail@web50301.mail.yahoo.com> Wendy, sounds like they were not processed correctlly, not sealed, or both. You can melt them down, go back through xylene 2 to 3 changes for 2 hours each, then go back to 100% alcohol for 2 to 3 changes for 2 hours each, then reprocess back to paraffin. FYI: Museums use a method of boiling in Weck soap to bring back mummy tissue. Had to use that method once when a tech left fresh breast sections on the xray tray overnight. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From contact <@t> excaliburpathology.com Tue Oct 12 11:49:27 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] eyes Message-ID: <20041012164927.57846.qmail@web50301.mail.yahoo.com> Bowman's membrane is in the cornea and can be demonstated with a standard periodic acid- Schiff stain. Avian eyes have an extra ring of bone around them that will need decalcification. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From jwatson <@t> gnf.org Tue Oct 12 11:49:53 2004 From: jwatson <@t> gnf.org (James Watson) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] alternates to OCT embedding media Message-ID: Kim, Thermo electron sells a m-1 embedding matrix, which is for -12 to -17 degrees. This works well on mouse brains because of the warmer temperature needed to section the tissue. OCT used to be sold for different temperatures and the optimum temperature for OCT now is below -17 degree. James Watson HT, ASCP Facilities Manager of Histology Genomics Institute of the Novartis Research Foundation 858-332-4647 jwatson@gnf.org -----Original Message----- From: Kim Merriam [mailto:kmerriam2003@yahoo.com] Sent: Tuesday, October 12, 2004 7:44 AM To: Histonet Subject: [Histonet] alternates to OCT embedding media Hello all, I was wondering if anyone knew of alternates to OCT-brand cryo-embedding media. I know that there are other brands of the stuff, but it is my understanding that they are all basically made out of the same thing. Does anyone know of any other media that is used to prepare frozen blocks? Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Express yourself with Y! Messenger! Free. Download now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Charles.Embrey <@t> carle.com Tue Oct 12 11:51:21 2004 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Pathologists' Assistants now certified by ASCP Message-ID: I just wanted to update the histonet on the new affiliation between the American Association of Pathologists' Assistants and the ASCP and BOR. Charles Embrey Pathologists' Assistant AAPA Histology Manager Carle Clinic, Urbana Illinois. For Immediate Release: Oct. 2, 2004 http://www.ascp.org/511live/Timssnet/News/TNT_news.cfm CHICAGO--The American Society for Clinical Pathology (ASCP) is pleased to announce that the American Association of Pathologists' Assistants (AAPA) will now be a participating member of the ASCP Board of Registry (BOR) Board of Governors. On Oct. 2, 2004, a Memorandum of Understanding (MOU) between the ASCP BOR and the AAPA was officially accepted and the two groups will begin offering the only professional certification for pathologists' assistants. AAPA Fellows are immediately eligible for ASCP certification. Beginning Oct. 1, 2005, other qualified candidates will be able to take the pathologists' assistant certification examination to become certified. Although the total number of individuals practicing as pathologists' assistants is unknown, most are probably AAPA members, each of whom had to pass a membership qualifying examination and meet strict training and experience criteria before becoming Fellows. According to Patricia Ellinger, MSEd, MT(ASCP)SBB, chair of the ASCP BOR Board of Governors, "Current AAPA Fellows can obtain certification through the BOR by submitting an application to the BOR between Oct. 1, 2004, and Dec. 31, 2005. After Dec. 31, 2005, all individuals seeking certification will have to pass the BOR certification examination to earn the credential. The certification will have multiple routes, similar to other BOR certifications. Examination applicants must be graduates of a pathologists' assistant educational program accredited by the National Accrediting Agency for Clinical Laboratory Science (NAACLS) or be able to demonstrate that they meet specific training and work experience requirements." National certification for pathologists' assistants is the realization of a long-held dream, explained AAPA member James W. Moore, MHS, who worked closely with the ASCP BOR to develop the MOU and establish the certification program. Until recently, he was the acting executive director of the National Commission on Certification of Pathologists' Assistants, a body initially organized by the AAPA to identify and evaluate various options for national certification. "In 1972 when the AAPA was formed, its primary purpose was to establish and maintain appropriate educational and professional standards for those working in the field and to seek national recognition and certification for its members," Moore said "Pathologists' assistants are physician extenders much like physicians' assistants," added AAPA Broad of Trustees Chair Thomas L. Reilly, BHS. "They are extensively trained and educated to provide complex anatomic pathology services under the supervision and direction of licensed, board-certified pathologists. It is not within their scope of practice to render a diagnosis," he stressed. "That is the responsibility of the pathologist." Ellinger expects the demand for pathologists' assistants to increase over time. "If the patient population continues to grow as expected, the need for laboratory services and pathologists' assistants will too." Reilly agrees with this view. "We are producing fewer pathologists than we used to, which is fueling the need for pathologists' assistants," he said. "And, I expect national certification to increase our visibility and possibly create additional demand." For more information or to find out how to apply for pathologists' assistant certification contact the ASCP BOR by e-mail at bor@ascp.org or call 1-800-621-4142, extension 1139. Application forms will be available after Oct. 15, 2004. As more information becomes available it will be posted on our website at www.ascp.org/bor, explained ASCP Senior Vice President and BOR Executive Director Kory Ward-Cook, Ph.D., MT(ASCP), CAE. # # # The American Society for Clinical Pathology (ASCP) is the largest association of pathologists and laboratory professionals in the world, with more than 140,000 members. ASCP is the leading provider of continuing education for pathologists and medical technologists and technicians, clinical scientists and other laboratory professionals. The ASCP's Board of Registry continues to certify the majority of the laboratory personnel working in the field and has certified more than 392,000 laboratorians. The ASCP promotes the health and safety of the public by enhancing the knowledge and skills of pathologists and laboratory professionals. For more information, visit the ASCP website at www.ascp.org. "America's Health Depends on Its Laboratories." From ttroyer <@t> pcllab.com Tue Oct 12 11:52:52 2004 From: ttroyer <@t> pcllab.com (Travis Troyer) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] lypmh node sections folding and falling off of immuno slides Message-ID: <004601c4b07b$e9dda6e0$6601010a@Peterson.local> I was wondering if there were any tricks to getting lymph nodes to remain flat and intact in order to do immuno stains. I have a problem with the tissue falling off or folding after the immuno run. Any thoughts or suggestions would be much appreciated. Thanks, Travis Troyer From cwscouten <@t> myneurolab.com Tue Oct 12 11:56:26 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] alternates to OCT embedding media Message-ID: Try Cryo M Bed from Vibratome company. OCT is softer than Cryo M Bed. See the link below: http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=475112&catdesc=Histology+Equipment&CatThreeID=671&CatOneID=4&subcatdesc=Cryostat+Accessories&idsubcategory=195 Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Merriam Sent: Tuesday, October 12, 2004 9:44 AM To: Histonet Subject: [Histonet] alternates to OCT embedding media Hello all, I was wondering if anyone knew of alternates to OCT-brand cryo-embedding media. I know that there are other brands of the stuff, but it is my understanding that they are all basically made out of the same thing. Does anyone know of any other media that is used to prepare frozen blocks? Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Express yourself with Y! Messenger! Free. Download now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Tue Oct 12 12:03:13 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Polyester membrane In-Reply-To: Message-ID: <001801c4b07d$5bf7e5f0$83020a0a@IHCTech> Has anyone ever used HISTOGEL as a freezing embedding media? Also, I was wondering what experiences if any you all have had with processing for sectioning polyester membranes with cells grown on them. Frozen, Paraffin, GMA????? I am securing the cut out membrane in histogel on edge and thinking of trying to gently paraffin process and/or freeze section??? Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of James Watson Sent: Tuesday, October 12, 2004 9:50 AM To: Kim Merriam; Histonet Subject: RE: [Histonet] alternates to OCT embedding media Kim, Thermo electron sells a m-1 embedding matrix, which is for -12 to -17 degrees. This works well on mouse brains because of the warmer temperature needed to section the tissue. OCT used to be sold for different temperatures and the optimum temperature for OCT now is below -17 degree. James Watson HT, ASCP Facilities Manager of Histology Genomics Institute of the Novartis Research Foundation 858-332-4647 jwatson@gnf.org -----Original Message----- From: Kim Merriam [mailto:kmerriam2003@yahoo.com] Sent: Tuesday, October 12, 2004 7:44 AM To: Histonet Subject: [Histonet] alternates to OCT embedding media Hello all, I was wondering if anyone knew of alternates to OCT-brand cryo-embedding media. I know that there are other brands of the stuff, but it is my understanding that they are all basically made out of the same thing. Does anyone know of any other media that is used to prepare frozen blocks? Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Express yourself with Y! Messenger! Free. Download now. _______________________________________________ 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 From Marjorie.Lehman <@t> unilever.com Tue Oct 12 12:03:03 2004 From: Marjorie.Lehman <@t> unilever.com (marjorie lehman) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] cut the old paraffin blocks Message-ID: Just curious - what is Weck soap? -----Original Message----- From: Paula Pierce [SMTP:contact@excaliburpathology.com] Sent: Tuesday, October 12, 2004 12:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cut the old paraffin blocks Wendy, sounds like they were not processed correctlly, not sealed, or both. You can melt them down, go back through xylene 2 to 3 changes for 2 hours each, then go back to 100% alcohol for 2 to 3 changes for 2 hours each, then reprocess back to paraffin. FYI: Museums use a method of boiling in Weck soap to bring back mummy tissue. Had to use that method once when a tech left fresh breast sections on the xray tray overnight. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Tue Oct 12 12:13:01 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] bone decalcification In-Reply-To: <20041012103337.5060C3326BD@perceval.uca.es> Message-ID: <001c01c4b07e$ba7d7ad0$83020a0a@IHCTech> Jose, If you are thinking of doing IHC Formic Acid is a good decal to use (5%). Decal Chemicals sells a formic acid product called Immunocal. There are other quicker products that I believe use Nitric acids but IHC is better with the Formic in my hands. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jose Luis Palazon Fernandez Sent: Tuesday, October 12, 2004 3:34 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] bone decalcification Dear List-members I have to dacalcify some fish vertebrae in order to see the vertebral articulation and intervertebral discs, and have to use a quieck method. I usually decalcify with EDTA but this method is very slow. I would appreciate your advise. Thanks in advance! Jos? Luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Oct 12 12:16:58 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] OCT purchase for those who cannot use Cardinal In-Reply-To: References: Message-ID: <6.0.0.22.1.20041012111612.01b3b738@gemini.msu.montana.edu> We order from VWR Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From pruegg <@t> ihctech.net Tue Oct 12 12:16:33 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Collagen IV antibody In-Reply-To: <20041012111946.SM05496@crgcsls814> Message-ID: <001d01c4b07f$391cf230$83020a0a@IHCTech> Lu you can try the U of Iowa Hybridoma Bank they have many anti-collagens http://www.uiowa.edu/~dshbwww/ -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Young Kwun Sent: Monday, October 11, 2004 6:24 PM To: 'Lu Leach'; Histonet Subject: RE: [Histonet] Collagen IV antibody Lu, You can get anti-Collagen IV, alpha 5 chain (Clone A7, Cat No: MC-885) from KAMIYA BIOMEDICAL COMPANY, Seatle, WA. www.kamiyabiomedical.com They also have alpha 1 and alpha 3 chains as well. Young Young Kwun Senior Hospital Scientist Dept of Anatomical Pathology Concord Hospital Concord NSW 2139 Australia Phone:61-2-9767 6075 Fax:61-2-9767 8427 Email:kwuny@email.cs.nsw.gov.au -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lu Leach Sent: Friday, 8 October 2004 6:37 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Collagen IV antibody Dear Histonetters, Can anyone tell me where to acquire Collagen IV A5 antibody? Thanks, Lu Leach University of Illinois at Chicago Research Pathology Core 312-996-3869 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "This message is intended for the addressee named and may contain confidential information. If you are not the intended recipient, please destroy it and notify the sender. Views expressed in this message are those of the individual sender, and are not necessarily the views of the Central Sydney Area Health Service." _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ploykasek <@t> phenopath.com Tue Oct 12 12:20:13 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] lymph node sections folding and falling off of immuno slides In-Reply-To: <004601c4b07b$e9dda6e0$6601010a@Peterson.local> Message-ID: Travis - I'll try to help, but need a little more info. What is the thickness of your sections? Are you using some type of adhesive slides? Does this only happen with heat pretreatment? If you'll answer these questions, we can have a better idea of possible solutions. Patti Loykasek PhenoPath Laboratories Seattle, WA> I was wondering if there were any tricks to getting lymph nodes to remain flat > and intact in order to do immuno stains. I have a problem with the tissue > falling off or folding after the immuno run. Any thoughts or suggestions > would be much appreciated. > Thanks, > Travis Troyer > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Angel.Enniss <@t> pfizer.com Tue Oct 12 12:21:34 2004 From: Angel.Enniss <@t> pfizer.com (Enniss, Angel) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Hematoxylin Message-ID: Our lab has been seeing some blue residue caused from Hematoxylin on our slides. It is not awful but we would like to try and eliminate it if possible. We are using Anatech Hematoxylin right now. Does anyone have a good hematoxylin that may leave cleaner slides? Thank you Angel LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From KTennako <@t> odu.edu Tue Oct 12 12:39:13 2004 From: KTennako <@t> odu.edu (Kushan U Tennakoon) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] microscope eyepiece adaptor Message-ID: We need to urgently purchase a microscope eyepiece adaptor to fix a Nikon Cool Pix Digital Camera (990) on to a Lieca microscope (Leica DMR) Wetzler Gmbh (Type 020-525-024 )in our lab. I would very much appreciate to know whether any one knows from where can I get a suitable eye piece adaptor Sincerely, Kushan Tennakoon ----------------------------------------------------------- Dr. Kushan Tennakoon Department of Biological Sciences Old Dominion University 110, Mills Godwin Bldg., 45th Street, Norfolk, VA 23529 -0266 USA. Tel: + 757 683 3610 Fax: + 757 683 5283 Email: ktennako@odu.edu http://web.odu.edu/sci/biology/tennakoon.html From froyer <@t> bitstream.net Tue Oct 12 12:43:38 2004 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] alternates to OCT embedding media In-Reply-To: References: Message-ID: <416C17CA.8090108@bitstream.net> ALSO... OCT can be ordered directly from Sakura Finetek, if all else fails. Product Code (Sakura): "4583" 4-oz bottle Call (800) 725-8723 ~ Ford Ford M. Royer, MT(ASCP) NAtional Sales Manager Midwest Science Biocenter Minneapolis, MN 55301 (800) 745-4869, Ext. 154 GUTIERREZ, JUAN wrote: >I believe they're called Cardinal these days, use to be Baxter and Allegiance for a while. The phone number I have is for Baxter, but I think it still works. 1-800-964-5227 > > >-----Original Message----- >From: Atoska S. Gentry [mailto:gentras@vetmed.auburn.edu] >Sent: Tuesday, October 12, 2004 10:30 AM >To: Histonet >Subject: Fwd: Re: [Histonet] alternates to OCT embedding media > > > >Hello, please who is your source for OCT these days??? Thanks, Atoska > From vazquezr <@t> ohsu.edu Tue Oct 12 13:03:50 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] lypmh node sections folding and falling off of immuno slides Message-ID: Travis, Try incubating overnight in a 37c oven. That might help. Are you cutting them at 3 microns? Robyn OHSU Portland, OR >>> "Travis Troyer" 10/12/2004 9:52:52 AM >>> I was wondering if there were any tricks to getting lymph nodes to remain flat and intact in order to do immuno stains. I have a problem with the tissue falling off or folding after the immuno run. Any thoughts or suggestions would be much appreciated. Thanks, Travis Troyer _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Tue Oct 12 13:08:35 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Hematoxylin Message-ID: Angel, After the hematoxylin and then the water, are you dipping them in 1% acid alcohol a couple of times to decolorize them? Robyn OHSU Portland,OR >>> "Enniss, Angel" 10/12/2004 10:21:34 AM >>> Our lab has been seeing some blue residue caused from Hematoxylin on our slides. It is not awful but we would like to try and eliminate it if possible. We are using Anatech Hematoxylin right now. Does anyone have a good hematoxylin that may leave cleaner slides? Thank you Angel LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GauchV <@t> mail.amc.edu Tue Oct 12 13:14:50 2004 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Gross Photography Photo Stand Message-ID: Does anyone know where I can purchase a good photo stand for taking pictures of specimens in the Grossing Lab? We have one from Lord knows how many years ago upstairs in our Grossing Room but now we need an additional one for our Frozen Section Grossing Room and are having a hard time finding anything to fit our needs. We have a digital camera and we would like to have a stand that iincorporates lighting as well as a holder for the camera that we could move up and down depending on the size of the specimen to keep the camera steady. The old one we have I think was something someone made at some point in time many moons ago..it has a plexiglas surface to put the specimen on with lights clamped on it and an adjustable arm for the camera above the plexiglas. If anyone has any sources for this type of stand, please let me know...Thanks in advance for your help... Vicki Gauch AMCH Albany, NY From DAKOTechServ <@t> dakocytomation.com Tue Oct 12 13:20:05 2004 From: DAKOTechServ <@t> dakocytomation.com (DAKOTechServ) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] DAKO Handbook on DAKO website Message-ID: <349C15307203B445A6B06AA6BA3D425F04AD5150@exchange.caus.dako.net> Dear Amos and the Histonet, Here is the address to DakoCytomation's Handbook. http://www.dakocytomation.us/index/us_support_literature_index/us_support_li terature_immhandbook.htm If you would like to request a copy to be mailed to you, please send an email to litreq@dakocytomation.com or call 800-400-3256 ext. 5242. Sincerely, Danielle Mach Technical Support Specialist DakoCytomation, Carpinteria, CA, USA 800-424-0021 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of amosbrooks@earthlink.net Sent: Tuesday, October 12, 2004 9:15 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] DAKO Handbook on DAKO website Gayle, When I first suggested the handbook I was going to include a link to it but when I went to the DAKO website to find it I found they had changed the site a lot since I was last there and I couldn't find the hand book. Do you happen to have a URL for it? I knew I should have bookmarked it when I saw it last! thanks, Amos Message: 12 Date: Mon, 11 Oct 2004 16:30:59 -0600 From: Gayle Callis Subject: [Histonet] DAKO Handbook on DAKO website To: Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20041011162938.01b2e378@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed You can download the Handbook from DAKOCYTOMATION website Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From victor <@t> pathology.washington.edu Tue Oct 12 13:25:18 2004 From: victor <@t> pathology.washington.edu (Victor Tobias) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] Gross Photography Photo Stand References: Message-ID: <416C218E.60009@pathology.washington.edu> Vicki, We are sort of in the same market. We were told of a interesting product, it is a light ring that fits around the lens. I personally have not seen it, but hear that it provides excellent pictures. Victor Vicki Gauch wrote: >Does anyone know where I can purchase a good photo stand for taking >pictures of specimens in the Grossing Lab? We have one from Lord knows >how many years ago upstairs in our Grossing Room but now we need an >additional one for our Frozen Section Grossing Room and are having a >hard time finding anything to fit our needs. We have a digital camera >and we would like to have a stand that iincorporates lighting as well as >a holder for the camera that we could move up and down depending on the >size of the specimen to keep the camera steady. The old one we have I >think was something someone made at some point in time many moons >ago..it has a plexiglas surface to put the specimen on with lights >clamped on it and an adjustable arm for the camera above the plexiglas. >If anyone has any sources for this type of stand, please let me >know...Thanks in advance for your help... > >Vicki Gauch >AMCH >Albany, NY > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax From pruegg <@t> ihctech.net Tue Oct 12 13:33:57 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] microscope eyepiece adaptor In-Reply-To: Message-ID: <002f01c4b08a$09223a30$83020a0a@IHCTech> Richard Kline of Microscope Services 303-905-0909 says he can make an adaptor for a digital camera for any scope. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kushan U Tennakoon Sent: Tuesday, October 12, 2004 10:39 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] microscope eyepiece adaptor We need to urgently purchase a microscope eyepiece adaptor to fix a Nikon Cool Pix Digital Camera (990) on to a Lieca microscope (Leica DMR) Wetzler Gmbh (Type 020-525-024 )in our lab. I would very much appreciate to know whether any one knows from where can I get a suitable eye piece adaptor Sincerely, Kushan Tennakoon ----------------------------------------------------------- Dr. Kushan Tennakoon Department of Biological Sciences Old Dominion University 110, Mills Godwin Bldg., 45th Street, Norfolk, VA 23529 -0266 USA. Tel: + 757 683 3610 Fax: + 757 683 5283 Email: ktennako@odu.edu http://web.odu.edu/sci/biology/tennakoon.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Tue Oct 12 13:46:24 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] microscope eyepiece adaptor Message-ID: Try Bioindustrialproducts.com. Page 6 of their catalog shows one that fits Leica microscopes, and I know they sell Nikon Coolpix cameras so they should know. Good luck. Talk to Mike Nedry, he is a real nice guy to deal with. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: Kushan U Tennakoon [mailto:KTennako@odu.edu] Sent: Tuesday, October 12, 2004 12:39 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] microscope eyepiece adaptor We need to urgently purchase a microscope eyepiece adaptor to fix a Nikon Cool Pix Digital Camera (990) on to a Lieca microscope (Leica DMR) Wetzler Gmbh (Type 020-525-024 )in our lab. I would very much appreciate to know whether any one knows from where can I get a suitable eye piece adaptor Sincerely, Kushan Tennakoon ----------------------------------------------------------- Dr. Kushan Tennakoon Department of Biological Sciences Old Dominion University 110, Mills Godwin Bldg., 45th Street, Norfolk, VA 23529 -0266 USA. Tel: + 757 683 3610 Fax: + 757 683 5283 Email: ktennako@odu.edu http://web.odu.edu/sci/biology/tennakoon.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From peoshel <@t> wisc.edu Tue Oct 12 12:59:00 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:24:09 2005 Subject: [Histonet] microscope eyepiece adaptor In-Reply-To: References: Message-ID: Thales-Optem, or whatever they're calling themselves nowadays: http://www.optemintl.com/DigitalCameraCouplers.html We have one for connecting a Nikon Coolpix 5000 to our various light microscopes. Works well, is well made, and is significantly cheaper than the other brand I've seen (which name I forget, but ...). Get the 30 mm accessory, and you'll be able to use this in most any microscope eyepiece or phototube, 22 mm or 30 mm diameter. But! This isn't enough! You also need to get the Nikon filter adapter. For the 5000, this was the E6-UR (if I remember right). This is a step-down adapter that screws into the front of the lens, and the eyepiece adapter then fits in the filter adapter. I don't know the part number for a 990. We bought ours from National Graphic Supply: http://www.ngscorp.com/science_medical.php But you'll have to call and talk to the rep for your area -- supplies like this are *not* obvious on their web site. Phil >We need to urgently purchase a microscope eyepiece adaptor to fix a Nikon >Cool Pix Digital Camera (990) on to a Lieca microscope (Leica DMR) Wetzler >Gmbh (Type 020-525-024 )in our lab. > >I would very much appreciate to know whether any one knows from where can I >get a suitable eye piece adaptor >Sincerely, > >Kushan Tennakoon >----------------------------------------------------------- >Dr. Kushan Tennakoon >Department of Biological Sciences >Old Dominion University >110, Mills Godwin Bldg., >45th Street, Norfolk, >VA 23529 -0266 >USA. >Tel: + 757 683 3610 >Fax: + 757 683 5283 >Email: ktennako@odu.edu >http://web.odu.edu/sci/biology/tennakoon.html > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From DRG <@t> Stowers-Institute.org Tue Oct 12 14:23:45 2004 From: DRG <@t> Stowers-Institute.org (Grant, Debra) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Research Technician II position available Message-ID: The Stowers Institute for Medical Research has an opening for a Laboratory Technician II to provide high quality support services in the Histology Facility. Responsibilities include receiving and processing specimens according to established protocol, researching information on currently used procedures and protocols, providing assistance on common-use equipment, maintaining equipment, and testing new procedures and conditions. In addition to excellent communication skills, and the ability to function independently as well as in a team-oriented environment, the successful candidate will have one year of experience in histopathology or in an EM lab within the last five years under the supervision of a pathologist or an appropriately certified medical scientist, and be HT(ASCP) certified or certification eligible. Minimum requirements include an AA degree with at least 12 semester hours in biology and chemistry, and experience in paraffin and frozen microtomy, routine and special stains. Previous experience with immunohistochemistry, in situ hybridization, methacrylate processing, and/or electron microscopy processing beneficial. Previous research experience helpful. To apply visit www.stowers-institute.org and click on "positions available" for instructions on submitting your resume. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From gcallis <@t> montana.edu Tue Oct 12 14:46:05 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Hematoxylin In-Reply-To: References: Message-ID: <6.0.0.22.1.20041012132811.01b2f560@gemini.msu.montana.edu> Angel, You did not say which hematoxylin you are using. Harris, Gill, another progressive formulation i.e. Anatechs hematoxylin?? Blue residue is caused by several things, the worst source is too much adhesive in a waterbath IF you use a gelatin or egg albumin based section adhesive. Plus charge slides can also have the problem. Really old hematoxylin (outdated) can also create some ugly problems including in tissue section. If you are using progressive Hematoxylin i.e. Gills formulations, the background staining on slide is removed very simply with a clarifying rinse after hematoxylin. Stain in hematoxylin running tap water 1 min 30 dips or so in 4% acetic acid (Richard Allen (RA) calls it Clarifier but it does contain acetic acid and RA recommends 1 minute in their clarifier). You don't want to leave sections in this solution for a long time, but some do the rinse for 30 seconds. Whatever works, it is not a differentiation step but rather a way to reduce background, break up ionic interactions of dye to slides. ANATECH should have recommendations for their product. Running tap water 1 min Bluing (Scotts tap water substitute or RA bluing solution) for 1 minute Rinse with running tap water 1 minute. Use fresh clarifier and bluing solutions daily in order to keep things clean and adjustment of sections to proper pH before eosin counterstaining. These solutions are cheap compared to having problems and doing recuts, or just ugly slides. Use adequate rinsing and if you use standing rinses, change these between slide racks. If you use Harris hematoxylin, the acid alcohol step should remove background from slides (unless excessive adhesive is on slides!). I don't think it is Anatech's hematoxylin that is the problem but rather other factors needed to fine tune your staining. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From sluhisto <@t> yahoo.com Tue Oct 12 14:58:44 2004 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] calcaflour white Message-ID: <20041012195844.55118.qmail@web51004.mail.yahoo.com> Anyone out there have a protocol or any information on doing calcaflour white for fungus on paraffin embedded human tissue??? Any help would be appreciated. Thank you. Susan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From gcallis <@t> montana.edu Tue Oct 12 15:09:27 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] acid alcohol versus acetic acid rinse, calling on Gary Gill to comment In-Reply-To: References: Message-ID: <6.0.0.22.1.20041012135116.01af32a8@gemini.msu.montana.edu> Robyn and others, You wrote: After the hematoxylin and then the water, are you dipping them in 1% acid alcohol a couple of times to decolorize them? ******************************************************************************************************************************* One needs to be careful here. When one says they use acid alcohol, they need to say what hematoxylin (name please) they use i.e. either progressive or regressive methods to avoid confusion. The only time we use 1% hydrochloric acid/alcohol mixture is after Harris's hematoxylin and then to "differentiate" or remove some of the hematoxylin stain from the nuclei to make it crisp and delicate, i.e.a regressive hematoxylin stain protocol. I have no doubt it could be used with progressive, but have rarely seen this done with concern this stronger acid could remove too much of the progressive type hematoxylin. Hopefully Gary Gill will comment on this. With progressive hematoxylins, Gill (half oxidized hematoxylin) including Richard Allan and other vendors formulations available, one does not require "differentiation" with an acid alcohol as these hematoxylins are not designed (hmm correct term?) to "overstain" the nuclei. We have always used a mild acid rinse aka clarifying rinse with 4% acetic acid to remove background from the slide surface with any progressive hematoxylin formulation. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Jenny-Oblander <@t> omrf.ouhsc.edu Tue Oct 12 15:12:20 2004 From: Jenny-Oblander <@t> omrf.ouhsc.edu (Jenny Oblander) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] calcaflour white Message-ID: Try www.doctorfungus.org . Jenny -----Original Message----- From: Histology SLU [mailto:sluhisto@yahoo.com] Sent: Tuesday, October 12, 2004 2:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] calcaflour white Anyone out there have a protocol or any information on doing calcaflour white for fungus on paraffin embedded human tissue??? Any help would be appreciated. Thank you. Susan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Tue Oct 12 15:14:52 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Canned statements for ASR antibodies Message-ID: Greetings Histo land can anyone direct me to a web page or other "official" site to get the wording on the immuno reagents labeled as ASR? Yep, you guessed it, preparing for a CAP inspection and yes, this is the first time the ASR issue came up and I know who my inspector is so this will be addressed during the inspection. Thank you so much. Oh, one other thing- How Bout Them New England Patriots!!!!!!!!!!!!! Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX From sbxjsr <@t> nottingham.ac.uk Tue Oct 12 15:50:13 2004 From: sbxjsr <@t> nottingham.ac.uk (Jan Roger) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] In situ detection problems Message-ID: I am having serious problems with my RNA:RNA in situ hybridisation. A couple of weeks ago, my result changed from having wonderful blue staining with NBT/BCIP, no background, to getting dirty brown slides with black precipitate, and much less staining. I have tried everything to avoid this problem including changing all buffers, baking glassware again etc., but no success (I have not tried changing antibody/ colour reagents, but different probe makes no difference). What am I missing? It cannot be a problem with my protocol in general, as it was working before. Is it a problem with tissue, detection reagents, antibody or probe? Any suggestions much appreciated. A brief run-down of my protocol (buffer washes not included) 4 % paraformaldehyde (PFA)-fixed paraffin-embedded sections, rehydrated through alcohols. 20 ug/ml Proteinase K to remove cross-links Post-fixation 5 min in 4 % PFA 10 min in 0.5 % acetic anhydride to acetylate sections Hyb buffer: 4 x SSC 40 % deionized formamide 1x Denhardts 250 ug/ml ssDNA 2 mM EDTA 37oC overnight Post-hyb washes: 2xSSC (2x15 min) 1xSSC (2x15 min) 0.1xSSC (2x30 min) 47oC Antibody: 1 % Roche blocking reagent + 2% normal serum +1:200 anti-DIG-AP Detection: 0.1 M Tris-HCl 0.1 M NaCl 0.05 M MgCl2 BCIP/NBT at manufacturers recommended dilution. Overnight 4oC Jan Roger Division of Animal Physiology Department of Biological Sciences University of Nottingham Sutton Bonington Campus Loughborough Leicestershire LE12 5RD Tel: 0115 9518862 Fax: 0115 9516302 This message has been scanned but we cannot guarantee that it and any attachments are free from viruses or other damaging content: you are advised to perform your own checks. Email communications with the University of Nottingham may be monitored as permitted by UK legislation. From Debbie.Vigil <@t> ahss.org Tue Oct 12 15:54:34 2004 From: Debbie.Vigil <@t> ahss.org (Vigil, Debbie (CTMC / CTPA)) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] nexes immuno & stainer Message-ID: <04Oct12.170636edt.119064@gateway.ahss.org> We have the old ventana nexes immuno and special stainer unit that we would like to sell. Does anyone know of a company that purchases this type of equipement? Thanks in Advance! CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From gcallis <@t> montana.edu Tue Oct 12 16:35:45 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Calcafluor white for fungus Message-ID: <6.0.0.22.1.20041012152741.01b3c9a0@gemini.msu.montana.edu> I know there was an excellent publication on calcafluor white in Laboratory Medicine, many years ago probably 80's. If you can access archives of this publication, official one from ASCP/CAP, and use key words - you should have the method. Not sure it can be accessed via PUBMED, worth a try. I no longer have a copy of this, but do recall journal and approx year(s). Someone used to have a kit for this purpose, PolySciences rings a distant bell. Contact Pam Marcum, she may know pmarcum@polysciences.com Good luck Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From AnthonyH <@t> chw.edu.au Tue Oct 12 17:06:46 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] acid alcohol versus acetic acid rinse, calling on Gary Gill to comment Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E0D1@simba.kids> Is the precipitate alum? Old solutions often leave a precipitate similar to fine blue tissue paper (the alum component). Haematein dye precipitate appears as dark blue granular dust on the sections. Usually a filter of the haematoxylin solution will solve this. You will probably find that if your haematoxylin is too old the nuclei will begin to appear brown. Regards, Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Wednesday, 13 October 2004 6:09 AM To: Robyn Vazquez; Histonet@lists.utsouthwestern.edu Subject: [Histonet] acid alcohol versus acetic acid rinse, calling on Gary Gill to comment Robyn and others, You wrote: After the hematoxylin and then the water, are you dipping them in 1% acid alcohol a couple of times to decolorize them? **************************************************************************** *************************************************** One needs to be careful here. When one says they use acid alcohol, they need to say what hematoxylin (name please) they use i.e. either progressive or regressive methods to avoid confusion. The only time we use 1% hydrochloric acid/alcohol mixture is after Harris's hematoxylin and then to "differentiate" or remove some of the hematoxylin stain from the nuclei to make it crisp and delicate, i.e.a regressive hematoxylin stain protocol. I have no doubt it could be used with progressive, but have rarely seen this done with concern this stronger acid could remove too much of the progressive type hematoxylin. Hopefully Gary Gill will comment on this. With progressive hematoxylins, Gill (half oxidized hematoxylin) including Richard Allan and other vendors formulations available, one does not require "differentiation" with an acid alcohol as these hematoxylins are not designed (hmm correct term?) to "overstain" the nuclei. We have always used a mild acid rinse aka clarifying rinse with 4% acetic acid to remove background from the slide surface with any progressive hematoxylin formulation. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From Mplhisto <@t> aol.com Tue Oct 12 18:35:28 2004 From: Mplhisto <@t> aol.com (Mplhisto@aol.com) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Re: Bile Message-ID: <8.593346f5.2e9dc440@aol.com> I was wondering if anyone would like to share thier procedure for preparing bile fluid for crystals. How long is the specimen good , what to do if the specimen is too thick,any tips you have would be greatly appreciated.Also someone mentioned to me they are running a control with it also, please let me know!! Thanks in advance. From CrochiereSteve <@t> aol.com Tue Oct 12 20:56:34 2004 From: CrochiereSteve <@t> aol.com (CrochiereSteve@aol.com) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] IHC for H.Pylori Message-ID: I have used Cell Marque's on the Ventana ES and am currently using Biocare's H.Pylori antibody on the Nemesis. Both work well. Steven M. Crochiere, HT(ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center Springfield, MA 01104 From contact <@t> excaliburpathology.com Tue Oct 12 21:05:33 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] calcafluor white Message-ID: <20041013020533.4356.qmail@web50301.mail.yahoo.com> Polysciences has a kit for fungus using calcofluor white. http://www.polysciences.com/shop/assets/datasheets/316.pdf You will need Adobe reader to view the page, otherwise just go to polysciences.com Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From contact <@t> excaliburpathology.com Tue Oct 12 21:27:01 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] cut the old paraffin blocks Message-ID: <20041013022701.89025.qmail@web50310.mail.yahoo.com> Sorry, Weck soap is a brand of liquid detergent. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From bhewlett <@t> cogeco.ca Tue Oct 12 15:50:56 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] acid alcohol versus acetic acid rinse, calling on Gary Gill to comment References: <6.0.0.22.1.20041012135116.01af32a8@gemini.msu.montana.edu> Message-ID: <00bb01c4b09d$2c4c52d0$6400a8c0@mainbox> Gayle and others, With either acid, 1% acid alcohol or 4% aqueous acetic acid, what you are doing is "acid differentiation"! The use of terms such as 'Clarifier" is nothing but obfuscation!!!!!! It doesn't matter if the hematoxylin solution is used progressively or regressively, acid rinses will still do two things; a) change the colour to red (hematoxylin is an indicator after all) and b) destabilize the dye lake attachment to the chromatin and progressively remove it. The degree of both things happening is related to the pH and the time of exposure. I must also point out that a couple of dips in 1% acid alcohol will not "decolourize" the section. Decolourization is the complete removal or loss of colour(see Oxford dictionary). The use of the term decolourize should be restricted to these occasions and not used to describe differentiation. OK, rant over. Irascible old guy, Bryan ----- Original Message ----- From: "Gayle Callis" To: "Robyn Vazquez" ; Sent: Tuesday, October 12, 2004 4:09 PM Subject: [Histonet] acid alcohol versus acetic acid rinse, calling on Gary Gill to comment > Robyn and others, > > You wrote: > > After the hematoxylin and then the water, are you dipping them in 1% acid > alcohol a couple of times to decolorize them? > **************************************************************************** *************************************************** > > One needs to be careful here. When one says they use acid alcohol, they > need to say what hematoxylin (name please) they use i.e. either progressive > or regressive methods to avoid confusion. The only time we use 1% > hydrochloric acid/alcohol mixture is after Harris's hematoxylin and then to > "differentiate" or remove some of the hematoxylin stain from the nuclei to > make it crisp and delicate, i.e.a regressive hematoxylin stain protocol. I > have no doubt it could be used with progressive, but have rarely seen this > done with concern this stronger acid could remove too much of the > progressive type hematoxylin. Hopefully Gary Gill will comment on this. > > With progressive hematoxylins, Gill (half oxidized hematoxylin) including > Richard Allan and other vendors formulations available, one does not > require "differentiation" with an acid alcohol as these hematoxylins are > not designed (hmm correct term?) to "overstain" the nuclei. We have > always used a mild acid rinse aka clarifying rinse with 4% acetic acid to > remove background from the slide surface with any progressive hematoxylin > formulation. > > > > > > > > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jkiernan <@t> uwo.ca Tue Oct 12 23:18:15 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] calcaflour white References: Message-ID: <416CAC87.CC733603@uwo.ca> The fluorochrome you mean is probably calcofluor white M2R (CI 40622, CI Fluorescent brightener 28). The name "calcofluor white" with various suffixes is used for many of the fluorescent whitening agents that are used to optically counteract the tendency of white fabrics to become yellow with age. These compounds are included in detergents used for washing clothes, for example. They are not all chemically related and will not all work the same way if used as fluorescent stains. Check that the Colour Index number and name are those of calcofluor white M2R. This fluorochrome has many uses (see RW Horobin's Chapter 22, "Polyene dyes and fluorochromes" in the 10th edn of Conn's Biological Stains, 2002). A fairly recent paper on the use of calcofluor white M2R for staining fungi is by R ruchel & M Schaffrinski (1999) J. Clin. Microbiol. 37:2694-2696. They comment that this fluorochrome can crystallize at high pH and recommend another one called "blankophor-P", whose identity I cannot determine. It's generally unwise to use any staining reagent that you don't know and understand, and foolish to use one that nobody knows anything about. For a simple staining technique using calcofluor white M2R see J Aslanzadeh & PS Stelmach (1996) Infection 24:248-250. (Their method is for Pneumocystis carinii but ought to be OK also for fungal hyphae.) All methods using calcofluor white M2R are much the same and extremely simple: Immerse hydrated sections in a dilute aqueous solution (about 0.1 mg/ml) for a couple of minutes, rinse in a few changes of pure water, thoroughly air-dry, rinse in xylene, and coverslip with a non-fluorescent resinous mounting medium. Excite with near-UV. The fluorescence is blue. John Kiernan London, Canada. __________________________________________________ Histology SLU wrote on 2th October: > > Anyone out there have a protocol or any information on doing calcaflour > white for fungus on paraffin embedded human tissue??? Any help would be > appreciated. Thank you. > > Susan From ctsblack <@t> capeheart.uct.ac.za Wed Oct 13 03:59:04 2004 From: ctsblack <@t> capeheart.uct.ac.za (Melanie Black) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Staining Challenge Message-ID: Hi There Anybody up for an unusual question??? What can I use to stain the [OH] group of Poly Vinyl Alcohol? Regards Melanie -- Cardiovascular Research Unit Div. of Cardiothoracic Surgery Chris Barnard Building University of Cape Town Anzio Road Observatory 7925 Republic of South Africa Tel +27 21 406-6589 Cel +27 82 469-3352 Fax +27 21 448-5935 From lpwenk <@t> sbcglobal.net Wed Oct 13 04:14:32 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] H. pylori References: Message-ID: <009101c4b105$0e3195e0$aa29d445@domainnotset.invalid> Don't know if this would apply to IHC on bacteria but . . . any chance that some of the bacteria are dead or dying? If they are dead or dying, the cell wall has been compromised. I know that this interferes with the Gram stain, causing Gram + not to retain the blue/black color, and subsequently staining with the red dye, making it look like a Gram - bacteria. Romanowsky type stains (Giemsa, Diff Quik, etc.) may not stain the bacteria at all. We see this on specimens that have too many microorganisms (where crowding causes a competition for nutrients and some die) or where the patient has started a treatment (antibiotic, Peptobismo, etc.) and some of the bacteria are dying. If the cell wall is being compromised in some of the bacteria, maybe the "number" of antigen sites are fewer in these compromised bacteria than in the non-compromised bacteria. So what you are seeing may be due to a difference in antibody-antigen binding sites, and thus a difference in staining intensity. Maybe try a Gram stain (Brown & Hopps, for example) or a Giemsa/Diff-Quik, so see if there is cell wall compromising going on. I'm not an expert on IHC. Just a thought. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Angela Bitting" To: Sent: Tuesday, October 12, 2004 7:40 AM Subject: [Histonet] H. pylori > We're perplexed. We've been using Dako's H. pylori AB with Proteinase K > antigen retreival for years now and have never had this problem. Our > control is teeming with organisms but some are staining brown and others > are more of a bluish color. I'm sure they are all H. Pyl organisms, but > I can't figure out why they aren't all staining the same. > Dako says they haven't had any complaints about their Lot number. We've > tried different vials from that same Lot # and both act the same. > Trying different dilutions didn't help. Now, we're extending the > Proteinase K time. After this, we're out of ideas. > Help! > > Angela Bitting, HT(ASCP) > Technical Specialist, Histology > Geisinger Medical Center 23-00 > 100 N Academy Ave. > Danville, PA 17822 > phone 570-214-9634 > fax 570-271-5916 > > > IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Andrew.Prior <@t> Smith-Nephew.com Wed Oct 13 05:38:34 2004 From: Andrew.Prior <@t> Smith-Nephew.com (Prior, Andrew) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Exakt System Message-ID: My lab is about to buy the Exakt system for resin embedding and cutting/grinding. Has anyone out there used this before? Any major problems? Hints and tips on use and on staining sections cut on it will be greatly appreciated. Possibly best to contact me directly with tips and I'll post a summary at a later date. Plus thanks for the info on the Dako Handbook - it sounds as if it is just what I need. Thanks All. Andrew Andrew Prior Histologist Smith & Nephew Research Centre York Science Park Heslington York Andrew.Prior@smith-nephew.com Confidentiality. This electronic transmission is strictly confidential to Smith & Nephew and intended solely for the addressee. It may contain information which is covered by legal, professional or other privilege. If you are not the intended addressee, or someone authorised by the intended addressee to receive transmissions on behalf of the addressee, you must not retain, disclose in any form, copy or take any action in reliance on this transmission. If you have received this transmission in error, please notify the sender as soon as possible and destroy this message. From jluis.palazon <@t> icman.csic.es Wed Oct 13 06:33:58 2004 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] bone decalcification Message-ID: <20041013113358.8B666337B76@perceval.uca.es> Dear List-fellows I want to thank all of you that answered to my query about quick decalcification. best wishes Jos? Luis El dia 12/10/2004 19:13 usted envio el siguiente mensaje: >Date: 12 de Octubre de 2004 19:13:01 >From: "Patsy Ruegg" >Subject: RE: [Histonet] bone decalcification >To: jluis.palazon@icman.csic.es, histonet@lists.utsouthwestern.edu > > Jose, > If you are thinking of doing IHC Formic Acid is a good decal to use > (5%). Decal Chemicals sells a formic acid product called Immunocal. > There are other quicker products that I believe use Nitric acids but IHC > is better with the Formic in my hands. > Patsy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jose > Luis Palazon Fernandez > Sent: Tuesday, October 12, 2004 3:34 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] bone decalcification > > > Dear List-members > > > > I have to dacalcify some fish vertebrae in order to see the vertebral > articulation and intervertebral discs, and have to use a quieck method. > I usually decalcify with EDTA but this method is very slow. I would > appreciate your advise. > > > > Thanks in advance! > > > > Jos? Luis > > Universidad de Oriente-Isla Margarita-Venezuela > > actualmente en: Instituto de Ciencias Marinas de Andalucia > > Puerto Real, C?diz, Espa?a. > > email: jluis.palazon@icman.csic.es > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es From BWilliams <@t> Lifespan.org Wed Oct 13 08:12:02 2004 From: BWilliams <@t> Lifespan.org (Williams, Blanche) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] EM stats Message-ID: <85151E3A0A49DF498F39F19EE56FEA8D05780B09@lsexch2.lsmaster.lifespan.org> Does anyone have any figures as to how many EM diagnostics and or scanning cases a tech is capable of doing in a calendar year? It would be a standard 40 hour work week. Thank you From hymclab <@t> hyhc.com Wed Oct 13 08:22:22 2004 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Canned statements for ASR antibodies Message-ID: Joe, I got my information from the long version of the CAP checklist that I printed off the CAP website www.cap.org/apps/docs/laboratory_accreditation/checklists/checklistftp.html Between laboratory and accreditation is an underscore line that doesn't show up when I type it here. Underneath the CAP question about ASR's they have sample disclaimer information. Here is what I use following the CAP guidelines and the inspectors have like the completeness of it. In fact one inspector took a copy to make sure his labs disclaimer was as complete. Since my last inspection though we don't have anymore ASR's that we perform so I don't have to use it anymore. "This test was developed and its performance characteristics determined by [insert manufacturer]. It has not been cleared or approved by the U.S. Food and Drug Administration. The FDA has determined that such clearance or approval is not necessary. This test is used for clinical purposes. It should not be regarded as investigational or for research. This laboratory is certified under the Clinical Laboratory Improvement Amendments of the 1988 (CLIA) as qualified to perform high complexity clinical laboratory testing." Not all of this is required, it is only recommended by CAP (see the long version of the CAP checklist). We only added this comment on to the surgical pathology report when doing ASR's. Hope this helps. -----Original Message----- From: Joe Nocito [mailto:JNocito@Pathreflab.com] Sent: Tuesday, October 12, 2004 3:15 PM To: Histonet Subject: [Histonet] Canned statements for ASR antibodies Greetings Histo land can anyone direct me to a web page or other "official" site to get the wording on the immuno reagents labeled as ASR? Yep, you guessed it, preparing for a CAP inspection and yes, this is the first time the ASR issue came up and I know who my inspector is so this will be addressed during the inspection. Thank you so much. Oh, one other thing- How Bout Them New England Patriots!!!!!!!!!!!!! Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kaburns <@t> med.unc.edu Wed Oct 13 08:27:18 2004 From: kaburns <@t> med.unc.edu (Kim Burns) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Polyester membrane Message-ID: <000601c4b128$5d0c9390$0b2e1398@peds.med.unc.edu> Hi Patsy, On which type of membrane are the cells grown? A transwell collagen membrane, t-clear, milli cell, snap well? We process cells grown on these types of polyester membranes on a regular basis. We leave the membrane attached to the support and process as usual, the exception being that we do not use xylene on our processor. We use a substitute called slide brite. The xylene can be to harsh for the support. After processing we sandwich the membrane between layers of paraffin and then cut the membrane from the support. We then cut the membrane in half, stand it on end in a mold and make the paraffin block. The best membranes to work with are transwells. T-clears tend to separate from the paraffin making sectioning difficult and have a tendency to come off subbed slides taking the cells with them. Kimberlie Burns Cystic Fibrosis Research Center University of North Carolina Date: Tue, 12 Oct 2004 11:03:13 -0600 From: "Patsy Ruegg" Subject: [Histonet] Polyester membrane To: "'James Watson'" , "'Kim Merriam'" , "'Histonet'" Message-ID: <001801c4b07d$5bf7e5f0$83020a0a@IHCTech> Content-Type: text/plain; charset="us-ascii" Has anyone ever used HISTOGEL as a freezing embedding media? Also, I was wondering what experiences if any you all have had with processing for sectioning polyester membranes with cells grown on them. Frozen, Paraffin, GMA????? I am securing the cut out membrane in histogel on edge and thinking of trying to gently paraffin process and/or freeze section??? Patsy From amosbrooks <@t> earthlink.net Wed Oct 13 08:28:03 2004 From: amosbrooks <@t> earthlink.net (amosbrooks@earthlink.net) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] DAKO Handbook on DAKO website Message-ID: <6806943.1097674084111.JavaMail.root@thecount.psp.pas.earthlink.net> Danielle, Thanks for sharing that with us. I really appreciate it. (As proof of what a complete geek I am, I actually had the book downloaded on my PDA. I think I need a vacation or something.) Anyway, I really appreciate the update and I now have it bookmarked in case the question arises again. Much obliged, Amos Message: 13 Date: Tue, 12 Oct 2004 11:20:05 -0700 From: DAKOTechServ Subject: RE: [Histonet] DAKO Handbook on DAKO website To: "'amosbrooks@earthlink.net'" , histonet@lists.utsouthwestern.edu Message-ID: <349C15307203B445A6B06AA6BA3D425F04AD5150@exchange.caus.dako.net> Content-Type: text/plain Dear Amos and the Histonet, Here is the address to DakoCytomation's Handbook. http://www.dakocytomation.us/index/us_support_literature_index/us_support_li terature_immhandbook.htm If you would like to request a copy to be mailed to you, please send an email to litreq@dakocytomation.com or call 800-400-3256 ext. 5242. Sincerely, Danielle Mach Technical Support Specialist DakoCytomation, Carpinteria, CA, USA 800-424-0021 From lizchlipala <@t> premierhistology.com Wed Oct 13 08:52:46 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] membranes Message-ID: <000001c4b12b$ebe2dcc0$76d48a80@AMY> Patsy We routinely process organotypic cultures on membranes, the ones that we are most familiar with are from Mattek, (I think they are Millipore MillicellT CM cell culture inserts) we process just like regular tissue and our tissue processor has xylene on it. We use a 8mm biopsy punch to remove the culture and membrane from the culture insert, process it between biospsy sponges and cut in half and embed each halve on end. We have not experienced any problems with sectioning, occasionally there might be some separation of the membrane from the tissue, but not that often. I'll send you some images. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Histology Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 lizchlipala@premierhistology.com www.premierhistology.com Ship to Address: Premier Histology Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From aep10 <@t> cornell.edu Wed Oct 13 09:55:00 2004 From: aep10 <@t> cornell.edu (Anna Elisse Beaudin) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] IHC withquantum dots Message-ID: <1876.128.253.96.73.1097679300.squirrel@128.253.96.73> Hello, Has anyone ever done immunohistochemistry on tissue sections using primary (or secondary) antibodies conjugated to quantum dots? if so, do you think it would be possible to do multiple labeling using different colored quantum dots? thanks in advance for your help! Anna Beaudin Division of Nutritional Sciences Cornell University From Jill.Cox <@t> providence.org Wed Oct 13 09:56:58 2004 From: Jill.Cox <@t> providence.org (Cox, Jill) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Microscope Slide stamp Message-ID: <05424658AAA6D441BBAFD9D9D98780B7044F0CC2@lcmmsg02.ca.providence.org> Hello Netters, I am trying to find some sort of stamp I can purchase to stamp slides. i.e. numbers and names. Is there such a thing out there? I can't afford a slide labeler so I need a less expensive solution. Thanks in advance, Jill DISCLAIMER: This message is intended for the sole use of the addressee, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If you are not the addressee you are hereby notified that you may not use, copy, disclose, or distribute to anyone the message or any information contained in the message. If you have received this message in error, please immediately advise the sender by reply email and delete this message. From bill501 <@t> mindspring.com Wed Oct 13 10:30:50 2004 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] calcaflour white In-Reply-To: <416CAC87.CC733603@uwo.ca> References: <416CAC87.CC733603@uwo.ca> Message-ID: At 12:18 AM -0400 10/13/04, John Kiernan wrote: >"calcofluor white" This sounds like the stuff that keeps the Jem'Hadar alive. Perhaps, if you know a Vorta, they could provide you with more information. ;-) BB From Torie.Kindschy <@t> parkview.com Wed Oct 13 10:36:11 2004 From: Torie.Kindschy <@t> parkview.com (Torie Kindschy) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] oxalate crystals Message-ID: Hey there, Does anyone know of a procedure for oxalate crystals? If so, the info would be greatly appreciated. Thanks, Torie From Michael.Rice <@t> holy-cross.com Wed Oct 13 11:09:34 2004 From: Michael.Rice <@t> holy-cross.com (Rice, Michael) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Microscope Slide stamp Message-ID: <3BC92F29BE821745AB15E04C98EE028D6935F3@HCH2KMAIL.holy-cross.com> You might want to look at labeling software and order sheets of microscope slide labels and use a PC to label them Mike Rice Holy Cross Hospital Ft Lauderdale -----Original Message----- From: Cox, Jill [mailto:Jill.Cox@providence.org] Sent: Wednesday, October 13, 2004 10:57 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microscope Slide stamp Hello Netters, I am trying to find some sort of stamp I can purchase to stamp slides. i.e. numbers and names. Is there such a thing out there? I can't afford a slide labeler so I need a less expensive solution. Thanks in advance, Jill DISCLAIMER: This message is intended for the sole use of the addressee, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If you are not the addressee you are hereby notified that you may not use, copy, disclose, or distribute to anyone the message or any information contained in the message. If you have received this message in error, please immediately advise the sender by reply email and delete this message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- From JNocito <@t> Pathreflab.com Wed Oct 13 11:15:48 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Canned statements for ASR antibodies In-Reply-To: Message-ID: Once again you guys have been great. Thanks for all the responses. Bring on the Inspectors. Hey, I think I'll wear my target shirt that I wore in Toronto the day of the inspection. Joe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joe Nocito Sent: Tuesday, October 12, 2004 3:15 PM To: Histonet Subject: [Histonet] Canned statements for ASR antibodies Greetings Histo land can anyone direct me to a web page or other "official" site to get the wording on the immuno reagents labeled as ASR? Yep, you guessed it, preparing for a CAP inspection and yes, this is the first time the ASR issue came up and I know who my inspector is so this will be addressed during the inspection. Thank you so much. Oh, one other thing- How Bout Them New England Patriots!!!!!!!!!!!!! Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Oct 13 12:33:42 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Pizzolata method for calcium oxalate crystals In-Reply-To: References: Message-ID: <6.0.0.22.1.20041013112249.01b6abe8@gemini.msu.montana.edu> Are you talking about calcium oxalate crystals in tissues? If so Pizzolata's method for calcium oxalate. You can find method in Hrapchak and Sheehan's Theory and Practice of Histotechnology. Our positive control was canine kidney from animal that had ingested antifreeze. We also did a von kossa to distinguish calcium phosphate from calcium oxalate on a separate, adjacent section. If you just want to see oxalate crystals in bodily fluids (urine) they have unique characteristics/formation under microscopic examination - polarized? long time since I had to do this. We used do this exam via microscope as Med techs. At 09:36 AM 10/13/2004, you wrote: >Hey there, >Does anyone know of a procedure for oxalate crystals? If so, the info >would be greatly appreciated. Thanks, Torie > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From GDawson <@t> Milw.Dynacare.com Wed Oct 13 12:39:11 2004 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] H. Pylori Message-ID: I use the Dako H. Pylori pre-dilute here in my lab. Glen Dawson BS, HT & QIHC (ASCP) IHC Coordinator Milwaukee, WI From dudleyboy2682 <@t> hotmail.com Wed Oct 13 13:15:22 2004 From: dudleyboy2682 <@t> hotmail.com (Jeremy Lind) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Please Remove My Address Message-ID: from the histonet mailing list. thank you. _________________________________________________________________ Is your PC infected? Get a FREE online computer virus scan from McAfee® Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From victoria.spoon <@t> bassett.org Wed Oct 13 13:33:04 2004 From: victoria.spoon <@t> bassett.org (Spoon, Victoria) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Pathology trasncription Message-ID: <1D9402BE940DF64E965196B5162C800108DDA6@exchange.bassett.org> I am looking for pathology transcription service. Is anyone familiar with a group/company that specializes in pathology? Thanks, Victoria Spoon, Anatomic Pathology Manager Bassett Hospital Cooperstown, NY 13326 (607)547-6357 NOTICE OF CONFIDENTIALITY This electronic message, including attachments, is for the sole use of the named recipient and may contain confidential or privileged information protected by New York State, and Federal regulations. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. If you are not the intended recipient or have received this communication in error please contact the sender or email.security@bassett.org and destroy all copies of the original message. Thank you. From akbitting <@t> geisinger.edu Wed Oct 13 15:02:39 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] New employee training Message-ID: Hi Folks, I'm hoping someone out there will be willing to help me get started on a new project. The Histology lab I began working at recently has no formal training/orientation for new employees. The most recent hospital-wide employee satisfaction survey cited poor training for new employees as a major complaint in many departments. It seems that there are techs who refuse to share what they know with others. I don't know if it's a job security issue or just "knowlege is power" or whatever.... Anyway, I am trying to formulate a new employee orientation checklist for new Histotechs coming into the department. Does anyone out there have something like this and would they mind sharing it with me, just to give me an idea how in-depth I need to go. Thanks. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From pruegg <@t> ihctech.net Wed Oct 13 17:13:32 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] IHC withquantum dots In-Reply-To: <1876.128.253.96.73.1097679300.squirrel@128.253.96.73> Message-ID: <001201c4b171$e05da1f0$83020a0a@IHCTech> What are "quantum dots"? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anna Elisse Beaudin Sent: Wednesday, October 13, 2004 7:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC withquantum dots Hello, Has anyone ever done immunohistochemistry on tissue sections using primary (or secondary) antibodies conjugated to quantum dots? if so, do you think it would be possible to do multiple labeling using different colored quantum dots? thanks in advance for your help! Anna Beaudin Division of Nutritional Sciences Cornell University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Wed Oct 13 20:57:05 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] IHC withquantum dots References: <001201c4b171$e05da1f0$83020a0a@IHCTech> Message-ID: <416DDCF1.E273B85D@uwo.ca> Quantum dots are nanocrystals of a semiconductor (exact chemical identity seems to be trade secret). They may have a future in a new generation of tiny computers. Quantum dots fluoresce in a wide range of colours and can be conjugated to antibodies, avidin, phalloidin etc. You can buy conjugated reagents or kits to conjugate your own. The Quantum Dot Corporation's web site http://www.qdots.com/live/index.asp shows some pretty pictures of labelled cells. There is a recent paper about quantum dots in the J. Histochem. Cytochem. Here's the bibliographic information and abstract, from http://www.jhc.org/cgi/content/abstract/52/1/13 If your institution subscribes to the journal you'll be able to get the full text on line. --------- Rozalia Nisman, Graham Dellaire, Ying Ren, Ren Li and David P. Bazett?Jones (2004) Application of Quantum Dots as Probes for Correlative Fluorescence, Conventional, and Energy-filtered Transmission Electron Microscopy J Histochem Cystochem 52:13?18 Luminescent semiconductor quantum dots (QDs) are a new class of fluorescent label with wide-ranging applications for cell imaging. The electron density and elemental composition of these materials permit the extension of their use as probes in conventional electron microscopy (TEM) and energy-filtered TEM (EFTEM). Here we illustrate the feasibility of using streptavidin-conjugated QDs as TEM tags by labeling a nuclear protein on cell sections and obtaining correlative fluorescence and TEM data. We also show that QD probes can be employed in conjunction with immunogold for co-localization of proteins at the ultrastructural level. Furthermore, by obtaining cadmium elemental maps of CdSe/ZnS QDs distributed on a nuclear structure, we demonstrate the potential of QDs for co-localization of multiple proteins when used in combination with EFTEM. --------- A Google search for quantum dots brings up thousands of hits. Searching for "quantum dots" and histochemistry and cytochemistry brought up 60 hits, at least half of them in English and relevant. John Kiernan London, Canada. ----------------------------------------------- Patsy Ruegg wrote: > > What are "quantum dots"? > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anna > Elisse Beaudin > Sent: Wednesday, October 13, 2004 7:55 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] IHC withquantum dots > > Hello, > > Has anyone ever done immunohistochemistry on tissue sections using > primary (or secondary) antibodies conjugated to quantum dots? if so, do > you think it would be possible to do multiple labeling using different > colored quantum dots? thanks in advance for your help! > > Anna Beaudin > Division of Nutritional Sciences > Cornell University ------------------------------------------------------- From pengbw <@t> sjtu.edu.cn Thu Oct 14 02:20:32 2004 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] IHC withquantum dots Message-ID: <20041014072032.7124A111A405@sjtu.edu.cn> We have a fellow in our lab who worked on QDs (quantum dots). So I have learned that most QDs can be excitated under wave length about between 400-600nm. It means that you can use one certain wave length to excitate several different QDs which are distinctive in emission wave length (thus different in color). But the drawback is that you can not use two or more QDs in order to see colocalization such as nucleus and membrane molecules of the same cell. Baowei Peng 1954 Huashan Road Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China From rentonlf <@t> bru.wits.ac.za Thu Oct 14 03:27:42 2004 From: rentonlf <@t> bru.wits.ac.za (renton louise mrs) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Exakt System Message-ID: <1097742462.8a1513c0rentonlf@bru.wits.ac.za> Andrew, I would strongly recommend that you get hands-on training from Dr Donath in Exakt Germany (if you have not already organised to do so. My colleague learned a tremendous amount from him, and it was well worth the expense. Sincerely -----Original Message----- From: "Prior, Andrew" To: "'histonet@lists.utsouthwestern.edu'" Date: Wed, 13 Oct 2004 11:38:34 +0100 Subject: [Histonet] Exakt System My lab is about to buy the Exakt system for resin embedding and cutting/grinding. Has anyone out there used this before? Any major problems? Hints and tips on use and on staining sections cut on it will be greatly appreciated. Possibly best to contact me directly with tips and I'll post a summary at a later date. Plus thanks for the info on the Dako Handbook - it sounds as if it is just what I need. Thanks All. Andrew Andrew Prior Histologist Smith & Nephew Research Centre York Science Park Heslington York Andrew.Prior@smith-nephew.com Confidentiality. This electronic transmission is strictly confidential to Smith & Nephew and intended solely for the addressee. It may contain information which is covered by legal, professional or other privilege. If you are not the intended addressee, or someone authorised by the intended addressee to receive transmissions on behalf of the addressee, you must not retain, disclose in any form, copy or take any action in reliance on this transmission. If you have received this transmission in error, please notify the sender as soon as possible and destroy this message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? From gu.lang <@t> gmx.at Thu Oct 14 04:07:16 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] verhoeff-hematoxylin Message-ID: <002401c4b1cd$34335d50$eeeea8c0@server> Hi I am dealing with russel-movat-stain and with verhoeff-hematoxylin and have a question about the recipe. In all books I have looked through, there is no HCl added except in John Kiernans Histological Methods. In the Weigert's Hematoxylin we use also acidified ferric chlorid to stain nuclei. And I have learned that nuclei stain better in acid solution. Now I have tried to stain elastic fibers with Verhoeff (AFIP-recipe) and after 30-45 min there was only a brown shadow. Can anybody give me some hints, what was going wrong? Is there a difference, if the Jodin-solution is made of Potassium-iodat instead of Potassium-iodide? What about the acid in the solution? thank you Gudrun Lang From i_stain <@t> yahoo.com Thu Oct 14 05:11:54 2004 From: i_stain <@t> yahoo.com (Scott Mordue) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] FGF antibody Message-ID: <20041014101154.5039.qmail@web42006.mail.yahoo.com> Hi, Does anyone know a good FGF (Fibroblast growth factor) antibody that work well for IHC in FFPE (formalin-fixed, paraffin-embedded) tissues? If yes, who is the supplier? Thank you in advance Scott --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From srishan <@t> mail.holyname.org Thu Oct 14 08:24:12 2004 From: srishan <@t> mail.holyname.org (srishan@mail.holyname.org) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] recycling xylene and alcohols Message-ID: Hi Everyone, Thank you so much for your responses regarding this matter. I am still doing some trial runs before making a decision. Nirmala Srishan From Luis.Chiriboga <@t> med.nyu.edu Thu Oct 14 08:32:54 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] IHC withquantum dots In-Reply-To: <001201c4b171$e05da1f0$83020a0a@IHCTech> Message-ID: Hi all, Most simply put, quantum dots are inorganic semiconductor nanocrystals. They can be made from a variety of doped semiconductor materials but I believe the most common for biological applications are cadmium and selenium. The semiconductor formed, is a crystal of only a few thousand atoms that have electron holes that give quantum dots their unique optical properties. When the dot is excited with a beam of light, they emit light that is directly proportional to their size. As the nanocrystals become smaller they emit light of high energy (shorter wavelength) and vice versa. Even more importantly, they emit in a very narrow band, allowing them to be used in a variety of ways and in combination. The problem has always been that they are extremely insoluble because the quantum dot core is kept inside a protective inorganic shell of zinc or other hydrophobic material. In the early 90's many researchers started playing with ways to increase the solubility. I believe the current method uses a mixture of hydrophobic and hydrophilic polymer (along the lines of micelle) forming compounds. The result is stable enough to allow attachment of quantum dots to proteins and other molecules of biological interest. I hope this helps...... Luis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Wednesday, October 13, 2004 6:14 PM To: 'Anna Elisse Beaudin'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] IHC withquantum dots What are "quantum dots"? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anna Elisse Beaudin Sent: Wednesday, October 13, 2004 7:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC withquantum dots Hello, Has anyone ever done immunohistochemistry on tissue sections using primary (or secondary) antibodies conjugated to quantum dots? if so, do you think it would be possible to do multiple labeling using different colored quantum dots? thanks in advance for your help! Anna Beaudin Division of Nutritional Sciences Cornell University _______________________________________________ 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 From peoshel <@t> wisc.edu Thu Oct 14 08:41:24 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] IHC withquantum dots In-Reply-To: <416DDCF1.E273B85D@uwo.ca> References: <001201c4b171$e05da1f0$83020a0a@IHCTech> <416DDCF1.E273B85D@uwo.ca> Message-ID: If I may, I'd like to amplify John's comments -- this is the area our lab is working in. Not so much quantum dots, as labels for correlated light microscopy and electron microscopy, and multiple labels for EM. So naturally, we've looked into QDs. They're mostly made of cadmium-selenide and similar alloys. The composition isn't secret, but QD is (reasonably) close about how they make the dots. It's not particularly easy to make quantum dots (unlike colloidal gold and other metals), and involves organic reagents and non-aqueous solvents. Which can make the conjugation difficult (as opposed to gold, which is easy). A second problem is that the elements used -- e.g., CdSe -- are toxic, which causes problems with living systems obviously, and with the antibodies or other ligands people wish to stick onto the dots. I believe QD's dots aren't so much conjugated to proteins or whatever in the sense that gold particles are, but are covalently bound with small intermediate molecules. This enlarges the final particle + macromolecule size and so reduces the spatial resolution useful in EM, but probably is irrelevant in LM. Phil >Quantum dots are nanocrystals of a semiconductor >(exact chemical identity seems to be trade secret). >They may have a future in a new generation of >tiny computers. > >Quantum dots fluoresce in a wide range of colours >and can be conjugated to antibodies, avidin, >phalloidin etc. You can buy conjugated reagents >or kits to conjugate your own. >The Quantum Dot Corporation's web site >http://www.qdots.com/live/index.asp >shows some pretty pictures of labelled cells. > >There is a recent paper about quantum dots in the >J. Histochem. Cytochem. Here's the bibliographic >information and abstract, from >http://www.jhc.org/cgi/content/abstract/52/1/13 >If your institution subscribes to the journal you'll >be able to get the full text on line. > >--------- >Rozalia Nisman, Graham Dellaire, Ying Ren, Ren Li and David P. >Bazett?Jones >(2004) >Application of Quantum Dots as Probes for Correlative Fluorescence, >Conventional, and Energy-filtered Transmission Electron Microscopy >J Histochem Cystochem 52:13?18 > >Luminescent semiconductor quantum dots (QDs) are a new class of >fluorescent label with wide-ranging applications for cell imaging. The >electron density and elemental composition of these materials permit the >extension of their use as probes in conventional electron microscopy >(TEM) and energy-filtered TEM (EFTEM). Here we illustrate the >feasibility of using streptavidin-conjugated QDs as TEM tags by labeling >a nuclear protein on cell sections and obtaining correlative >fluorescence and TEM data. We also show that QD probes can be employed >in conjunction with immunogold for co-localization of proteins at the >ultrastructural level. Furthermore, by obtaining cadmium elemental maps >of CdSe/ZnS QDs distributed on a nuclear structure, we demonstrate the >potential of QDs for co-localization of multiple proteins when used in >combination with EFTEM. >--------- > >A Google search for quantum dots brings up thousands of hits. >Searching for "quantum dots" and histochemistry and cytochemistry >brought up 60 hits, at least half of them in English and >relevant. > >John Kiernan >London, Canada. >----------------------------------------------- >Patsy Ruegg wrote: >> >> What are "quantum dots"? >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anna >> Elisse Beaudin >> Sent: Wednesday, October 13, 2004 7:55 AM >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] IHC withquantum dots >> >> Hello, >> >> Has anyone ever done immunohistochemistry on tissue sections using >> primary (or secondary) antibodies conjugated to quantum dots? if so, do >> you think it would be possible to do multiple labeling using different >> colored quantum dots? thanks in advance for your help! >> >> Anna Beaudin >> Division of Nutritional Sciences >> Cornell University >------------------------------------------------------- > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From pruegg <@t> ihctech.net Thu Oct 14 09:57:01 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] FGF antibody In-Reply-To: <20041014101154.5039.qmail@web42006.mail.yahoo.com> Message-ID: <001d01c4b1fe$1115fe00$83020a0a@IHCTech> Scott back in the day I used FGF from Sigma on ffpe bone using pepsin digestion as pretreatment, I don't know if Sigma still carries the antibody. Santa Cruz sells a lot of growth factor antibodies also. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Scott Mordue Sent: Thursday, October 14, 2004 3:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] FGF antibody Hi, Does anyone know a good FGF (Fibroblast growth factor) antibody that work well for IHC in FFPE (formalin-fixed, paraffin-embedded) tissues? If yes, who is the supplier? Thank you in advance Scott --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Oct 14 09:58:14 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] IHC withquantum dots In-Reply-To: <001201c4b171$e05da1f0$83020a0a@IHCTech> References: <1876.128.253.96.73.1097679300.squirrel@128.253.96.73> <001201c4b171$e05da1f0$83020a0a@IHCTech> Message-ID: <6.0.0.22.1.20041014083855.01b23300@gemini.msu.montana.edu> Patsy and everyone, QDots are the hot new thing on the market, nanocrystals that can be conjugated to Strepavidin, antibodies and mnay othre things, i.e. proteins. They vary in size and are highly fluorescent, and are very pricey. The different nanocrystal sizes determine the final color that is emitted after excitation which is extremely bright and doesn't photobleach, a wonderful thing for those of us who work with confocals and fluorescence microscopes. You can learn more about them by going to www.qdots.com. The website has excellent explanations on QDots, a good place to learn about adsorption/emission, etc basic explanations on fluorescence. Science magazine has many publications in the past few years explaining their structure and applications in animals/tissue research, including breast cancer research. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From portera203 <@t> yahoo.com Thu Oct 14 10:21:02 2004 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] PPAR Gamma Questions Message-ID: <20041014152102.73744.qmail@web40913.mail.yahoo.com> Is anybody out there using PPar gamma 1 or PPar gamma 2 primaries on mouse tissue? If you are or know of any sources where we might be able to purchase them separately I would really appreciate the info. I am finding a lot of PPar Gamma 1&2 together - however we have a client who would like to use them individually. Any help would be appreciated. Thanks in advance, Amy S.Porter, HT(ASCP) Michigan State University Department of Physiology Division of Human Pathology College of Human Medicine portera203@yahoo.com --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From ASelf <@t> gmhsc.com Thu Oct 14 10:54:36 2004 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Histology Shipping Regulations Message-ID: <39836CD6DB61654E8F95A35898C921860A883D@exchange.gmhpost.com> Histonetters, How is everyone packaging their blocks/slides to be mailed out to other institutions, in reference to the new shipping regulations? Thanks in advance, amy Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or an agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. From ecl <@t> itsa.ucsf.edu Thu Oct 14 11:16:35 2004 From: ecl <@t> itsa.ucsf.edu (Ellen Liebenberg) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Staining rabbit mast cells Message-ID: <6.0.1.1.2.20041014085551.023e8760@itsa.ucsf.edu> I am trying to identify mast cells in rabbit peripheral nerve. The tissue is formalin fixed, postfiixed in osmium ( for myelin staining), and embedded in Technivit 7100. I have tried a variety of acidic Toluidine blue, alcian blue, safranin O, and thionin protocols, as well as an alkaline thionin protocol from Bancroft, all of which are supposed to identify mast cells. Areas of cartilage are staining as expected, but no mast cells. I am looking in the highly vascular connective tissue around the median nerve. Some of the specimens have been compressed and show myelin degeneration and extensive cellular reaction. Masts cells are probably present. I plan to process rabbit skin as a control tissue, but have not got the tissue yet. Could the osmium be interfering with staining? I have tried both non osmium stained gma and paraffin sections and still did not see mast cells. Do rabbit mast cells stain differently from other species? I plan to try the chloroacetate esterase kit from Sigma, but it might not work with osmium. Any suggestions would be greatly appreciated. Ellen Liebenberg University Califonia Medical Center San Franscisco, Ca. From katri <@t> cogeco.ca Thu Oct 14 11:18:02 2004 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] verhoeff-hematoxylin References: <002401c4b1cd$34335d50$eeeea8c0@server> Message-ID: <002001c4b209$619549c0$6a9a9618@Katri> Hi Gudrun, I'm not familiar with the russel-movat stain and I'm not quite sure what your question is pertaining to. The original Verhoeff's hematoxylin for elastic fibres does not use HCl, you are staining elastic fibres, not nuclei as with Weigert's hematoxylin, where HCl is added. Musto's modification of Verhoeff's in John Kiernan's book has HCl added and it does stain nuclei also. I don't think you should substitute potassium iodide with potassiun iodate, I'm sure somebody on the list will explain why. Also working solution is not stable and should be freshly prepared. I hope this will help... Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Gudrun Lang" To: "Histonetliste" Sent: Thursday, October 14, 2004 5:07 AM Subject: [Histonet] verhoeff-hematoxylin Hi I am dealing with russel-movat-stain and with verhoeff-hematoxylin and have a question about the recipe. In all books I have looked through, there is no HCl added except in John Kiernans Histological Methods. In the Weigert's Hematoxylin we use also acidified ferric chlorid to stain nuclei. And I have learned that nuclei stain better in acid solution. Now I have tried to stain elastic fibers with Verhoeff (AFIP-recipe) and after 30-45 min there was only a brown shadow. Can anybody give me some hints, what was going wrong? Is there a difference, if the Jodin-solution is made of Potassium-iodat instead of Potassium-iodide? What about the acid in the solution? thank you Gudrun Lang _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SohrabB1 <@t> wmmcpo.ah.org Thu Oct 14 11:22:52 2004 From: SohrabB1 <@t> wmmcpo.ah.org (Behnaz Sohrab) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Test Message-ID: From SohrabB1 <@t> wmmcpo.ah.org Thu Oct 14 11:31:46 2004 From: SohrabB1 <@t> wmmcpo.ah.org (Behnaz Sohrab) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Employment Message-ID: White Memorial Med. Ctr. In Los Angeles is seeking a full time certified Histotechnologist. Hours: Monday-Friday 3 am- 11.30 am From weneng2004 <@t> yahoo.com Thu Oct 14 11:52:30 2004 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Thanks!/cutting the old paraffin block Message-ID: <20041014165230.43649.qmail@web53407.mail.yahoo.com> Thank you all who advice me how to make sections from old paraffin blocks! As you taught, I re-embedded the sample by melting down the paraffin,xylene, EtOH,Xylene,paraffin, then embedding. Finally when I tested it on microtome, beautiful sections are produced! After that I sealed the samples with paraffin. Thank you very much! And thanks to histonet, I learned a lot here. Wendy --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From Jill.Cox <@t> providence.org Thu Oct 14 12:05:47 2004 From: Jill.Cox <@t> providence.org (Cox, Jill) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Thanks for slide label responses!! Message-ID: <05424658AAA6D441BBAFD9D9D98780B7044F0CC3@lcmmsg02.ca.providence.org> Thanks to all of you for the responses. I went to Staples and found a rubberstamp that letters can be changed for $22.00. Wow, what a find. I also ordered some chemical resistant ink and pad so now I am good to go. Thanks again, jill DISCLAIMER: This message is intended for the sole use of the addressee, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If you are not the addressee you are hereby notified that you may not use, copy, disclose, or distribute to anyone the message or any information contained in the message. If you have received this message in error, please immediately advise the sender by reply email and delete this message. From tpmorken <@t> labvision.com Thu Oct 14 12:16:10 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] IHC withquantum dots Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA257F5E2@usca0082k08.labvision.apogent.com> <> Actually, you can see several colors at once, depending on if their emissions fit into the same bandpass filter range. I've seen three color preparations that are spectacular. Tim Morken Lab Vision - Neomarkers www.labvision.com Free webhosting for US State Histotechnology Societies: http://www.labvisioncorp.com/demowebsite/index.cfm -----Original Message----- From: Baowei Peng [mailto:pengbw@sjtu.edu.cn] Sent: Thursday, October 14, 2004 12:21 AM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] IHC withquantum dots We have a fellow in our lab who worked on QDs (quantum dots). So I have learned that most QDs can be excitated under wave length about between 400-600nm. It means that you can use one certain wave length to excitate several different QDs which are distinctive in emission wave length (thus different in color). But the drawback is that you can not use two or more QDs in order to see colocalization such as nucleus and membrane molecules of the same cell. Baowei Peng 1954 Huashan Road Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China From dmccaig <@t> ckha.on.ca Thu Oct 14 12:14:06 2004 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] technician vs technologist Message-ID: <3E5A3F039F0BD8118B4700C00D002024043316@CKHA9> Other than frozen sections and special stains, are there any other restrictions a MLA or technician can not perform? Are they restricted to sectioning non biopsy cases or any if they prove their ability. With the trend to replace tech's with aids, I want to be sure of their capabilities. Diana McCaig, R.T. Charge Tech, Histology Chatham Kent Health Alliance 519-352-6401 (6604) From Richard.Breckenridge <@t> uth.tmc.edu Thu Oct 14 12:36:56 2004 From: Richard.Breckenridge <@t> uth.tmc.edu (Richard A Breckenridge) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] HCV In Situ Message-ID: Is anyone doing HCV In Situ? We are looking to do this and I would welcome any information as to products used and especially protocols performed. Thanks in advance. Richard A. Breckenridge Technical Director/ Chief Histology Technician UT-Houston Medical School Histology/ IHC Laboratory Office 713-500-6792 IHC 713-500-5096 Histology Lab 713-500-5363 Richard.Breckenridge@uth.tmc.edu From ahenn <@t> vaccinex.com Thu Oct 14 14:06:27 2004 From: ahenn <@t> vaccinex.com (Alicia Henn Ph.D.) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] quantum dots Message-ID: <001001c4b220$e8a1ec40$2c01a8c0@AHenn> Hi, This is a first-time post for me. Quantum dots (Qdots) are tiny fluorescent wafers similar to computer chips. The size of the dot determines the wavelength it emits and they are available in lots of different colors. While they excite at a broad range of wavelengths, you'll need different filters for optimum detection of the signal from the different colors. Look up the Quantum dot manual, available on their website, to see if you have the right filters for the two colors you'd like. http://www.qdots.com/live/upload_documents/90-0003.pdf Qdots are far brighter than any fluorescent dye. I've used their Streptavidin-655, but had lower background with my own primary antibody conjugated to the 655 Qdots. The conjugation is quick and easy with their kit. Alicia Henn, Ph.D. Vaccinex, Inc. Rochester, NY From Cathy.Stevens <@t> HealthONEcares.com Thu Oct 14 12:46:41 2004 From: Cathy.Stevens <@t> HealthONEcares.com (Stevens Cathy) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] New employee training Message-ID: We have a competency to be completed within the first 30 days of employment. It covers everything I could think of that is required of them. They only sign off when they feel comfortable with the procedure. There are different levels to the competency, some is just talking about the procedure, others include actual hands on/watching them perform. This is also done yearly. Hope it helps! -----Original Message----- From: Angela Bitting [mailto:akbitting@geisinger.edu] Sent: Wednesday, October 13, 2004 2:03 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] New employee training Hi Folks, I'm hoping someone out there will be willing to help me get started on a new project. The Histology lab I began working at recently has no formal training/orientation for new employees. The most recent hospital-wide employee satisfaction survey cited poor training for new employees as a major complaint in many departments. It seems that there are techs who refuse to share what they know with others. I don't know if it's a job security issue or just "knowlege is power" or whatever.... Anyway, I am trying to formulate a new employee orientation checklist for new Histotechs coming into the department. Does anyone out there have something like this and would they mind sharing it with me, just to give me an idea how in-depth I need to go. Thanks. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carl.hobbs <@t> kcl.ac.uk Thu Oct 14 14:22:58 2004 From: carl.hobbs <@t> kcl.ac.uk (carl hobbs) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] re QDs Message-ID: I?m very interested ..I am able to access a range that I haven?t yet checked out but the person who?s trailed them hasn?t had much success.what I would like to be clarified is what fluorescent filters are needed can one utilise the conventional ?FITC?, ?TRITC?, ?DAPI? filters? Apologies for my ignorance --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.762 / Virus Database: 510 - Release Date: 13/09/2004 From GREYTRUNK <@t> aol.com Thu Oct 14 15:08:49 2004 From: GREYTRUNK <@t> aol.com (GREYTRUNK@aol.com) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] technician vs technologist Message-ID: <1b8.37a58e5.2ea036d1@aol.com> This would also be dependent upon what your state reuirements are. I know here in Florida a technician cannot do IHC and Lab assistants CANNOT do anything technical. The lab assistants are designed to assistin non technical areas, ie. assisting with gross, accessioning, changing the processors, coverslipping, distributing slides, etc. Roxanne From KathyOconnor <@t> hmh.westsound.net Thu Oct 14 15:31:32 2004 From: KathyOconnor <@t> hmh.westsound.net (Kathy OConnor) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Microwave tissue processors Message-ID: We are looking to purcase a microwave tissue processor for late arriving biopsies. What has been the experience of users in Histoland as to preferred manufacturers? Kathy O'Connor Pathology Associates Bremerton, WA From Annette_hall <@t> pa-ucl.com Thu Oct 14 22:09:30 2004 From: Annette_hall <@t> pa-ucl.com (Annette Hall) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Nuclear Bubbles Message-ID: <9FC023A4AB52BB4D87DC6456081A822C070181@mercury.pa-ucl.com> Hi, Over the past 6 months, our lab has struggled intermittently with an artifact on our H&E slides. The nucleus will appear to have bubbled. This seems to be most prevalent in prostate biopsies, but we have also seen it in endometrial biopsies. We use a Leica Autostainer XL with Richard Allan reagents. We have adjusted drying times and oven temps but we still continue to occasionally see bubbles. Has anyone had this experience or know what is causing this? Any suggestions would be greatly appreciated. Thanks, Annette J. Hall, MT Micro/Histo/Cyto Supervisor United Clinical Labs 205 Bluff St. Dubuque, IA 52001 563.556.2010 x131 From lpwenk <@t> sbcglobal.net Fri Oct 15 01:20:30 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Nuclear Bubbles References: <9FC023A4AB52BB4D87DC6456081A822C070181@mercury.pa-ucl.com> Message-ID: <003f01c4b27f$12f8c320$922ad445@domainnotset.invalid> Tissues fixed in formalin are more susceptible to nuclear bubbling than a mercury or zinc fixative, due to formalin being a poor fixative for nuclei - it doesn't cross-link and stabilized the nuclear proteins as well as metal fixatives (zinc-formalin, mercuric fixatives such as Zenker or B5).. Tissues under-fixed in formalin are more susceptible to nuclear bubbling than those tissues properly fixed, as the nuclear proteins do not have as many stabilizing cross-links. Placing slides in drying ovens over 60 degrees C. are more likely to show nuclear bubbling than slides dried under 60 degrees C., especially those tissues under-fixed in formalin. The theory is that the water under the tissue "steams" at the higher temperatures, breaking the few cross-links in the nuclei, thus rearranging the chromatin around the outside of the steam bubbles. Well fixed tissue would have enough nuclear cross-links to resist the chromatin rearrangement by steam. Nuclei fixed with metal fixatives have more and stronger cross-links to resists the steam rearrangement than formalin fixed tissues. So look at fixation time and slide drying temperature. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Annette Hall" To: Sent: Thursday, October 14, 2004 11:09 PM Subject: [Histonet] Nuclear Bubbles > Hi, > > Over the past 6 months, our lab has struggled intermittently with an > artifact on our H&E slides. The nucleus will appear to have bubbled. This > seems to be most prevalent in prostate biopsies, but we have also seen it in > endometrial biopsies. We use a Leica Autostainer XL with Richard Allan > reagents. We have adjusted drying times and oven temps but we still continue > to occasionally see bubbles. Has anyone had this experience or know what is > causing this? Any suggestions would be greatly appreciated. > > Thanks, > Annette J. Hall, MT > Micro/Histo/Cyto Supervisor > United Clinical Labs > 205 Bluff St. > Dubuque, IA 52001 > 563.556.2010 x131 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carl.hobbs <@t> kcl.ac.uk Fri Oct 15 01:54:39 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] re QDS Message-ID: <001901c4b283$d761b060$e8345c9f@Carlos> Many thanks to Gayle and Tim for their very helpful advice From paivi.parkkisenniemi <@t> lshp.fi Fri Oct 15 05:15:10 2004 From: paivi.parkkisenniemi <@t> lshp.fi (=?iso-8859-1?Q?Parkkisenniemi_P=E4ivi?=) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] about flow-cytometry in small pathology labs Message-ID: <000a01c4b29f$dab6e520$5e0010ac@PATOLNEUVHUON> Hello, I?m asking if there is any small pathology unit that uses flow-cytometer ? I?m interested to hear about what manufacturer?s machine do you use, how much does it cost and any recommendatios you have. We are a small lab and looking for a cheap, effective flow-cytometer that is suitable for our laboratory.Thank you for your help in advance. P?ivi Parkkisenniemi medical.lab techologist Pathology laboratory Lapland Central Hospital Rovaniemi Finland From leclercj <@t> vetanat.unizh.ch Fri Oct 15 06:39:26 2004 From: leclercj <@t> vetanat.unizh.ch (Leclerc Jocelyne) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] silver stain with plastic embedded tendons Message-ID: Hi, has anyone experience with silver stain on thick slide of plastic embedded horse tendon which are about 150 micrometer? We want to stain with Bodian and Rager. The tendons are embedded in MMA and the fixation was with formalin and glutaraldehyd. Thank you for your help. Jocelyne Leclerc VetSuisse Fakult?t Veterin?r-Anatomisches Institut Winterthurerstr. 260 8057 Z?rich From Rcartun <@t> harthosp.org Fri Oct 15 09:00:50 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Histology Shipping Regulations Message-ID: What new shipping regulations? Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Amy Self" 10/14/04 11:54AM >>> Histonetters, How is everyone packaging their blocks/slides to be mailed out to other institutions, in reference to the new shipping regulations? Thanks in advance, amy Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or an agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. From vazquezr <@t> ohsu.edu Fri Oct 15 09:06:07 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Histology Shipping Regulations Message-ID: Amy, Is there an infection/contamination regulation on slide and blocks? This is the first I have ever heard of it and I ship slides all the time. Robyn OHSU Portland, Or >>> "Richard Cartun" 10/15/2004 7:00:50 AM >>> What new shipping regulations? Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Amy Self" 10/14/04 11:54AM >>> Histonetters, How is everyone packaging their blocks/slides to be mailed out to other institutions, in reference to the new shipping regulations? Thanks in advance, amy Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or an agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From David.Muskett <@t> RLC.NHS.UK Fri Oct 15 09:34:36 2004 From: David.Muskett <@t> RLC.NHS.UK (Muskett David) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Performance Indicators in Histopathology[Scanned] Message-ID: <6AB79F0BA7C4914BA8DD8F5E0C21B9752E25A6@AHEXMAIL01.xalderhey.com> Dear All I am inquiring if anyone has any experience of using performance indicators to assess the quality of their histopathology service. We are in all the appropriate EQA schemes have internal quality control systems but these do not measure the performance of the service as a whole. Turnaround times have always appeared as a very poor way to measure quality. If anyone has any ideas or experience or can guide me to any appropriate literature I would be grateful. Regards David David Muskett Chief Biomedical Scientist, Histology Royal Liverpool Children's NHS Trust Eaton Road Liverpool L12 2AP Internal telephone x3615 Telephone (0151) 293 3656 Fax (0151) 293 3617 http://nhsald02/histopathology From JWEEMS <@t> sjha.org Fri Oct 15 10:27:36 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Controls for Immunos fixed in B-Plus Message-ID: <83AACDB0810528418AA106F9AE9B7F7E2784D5@sjhaexc02.sjha.org> Hi Everyone, Do you use B-Plus fixed control tissue for immunos done on B-Plus fixed patient tissue? If so, how does it compare to formalin or B-5? Thanks in advance. j And happy weekend to everyone! j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. -------------- next part -------------- Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s Health System, Inc. From TMcNemar <@t> lmhealth.org Fri Oct 15 10:45:35 2004 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] unsubscribe Message-ID: <90092A4ED388D7119575006008F7112049CBE7@NT_EXCHANGE> -----Original Message----- From: HistoNet Server [mailto:histonet@pathology.swmed.edu] Sent: Thursday, March 09, 2000 9:20 AM To: Tom T. McNemar Subject: re: subscribe Your address has been added to the addresses that comprise this Listserv List. Welcome to HISTONET. This is an electronic mailing list for the exchange of information pertaining to histotechnology and related fields. PLEASE SAVE THIS MESSAGE. It contains useful information about how to use the list and what to do if you experience problems. It also includes some basic rules for email etiquette (Netiquette) which will be helpful to those who are new to this form of communication. WHAT IS A LISTSERVER? A list server is a computer that runs software which will receive incoming electronic mail (email) messages and reroute them automatically to everyone on the subscriber list. Email uses the vast expanse of the Internet to allow almost instantaneous communication between networked computers around the world. Our system uses the LISTSTAR software from Quarterdeck Corporation (California) and can currently send about 30 messages a minute. With the present number of subscribers, we are processing about 10,000 outbound messages a day. WHO SHOULD SUBSCRIBE? Anyone interested in research or clinical applications of histology, immunohistochemistry, in-situ hybridization pathology, and electron microscopy may find Histonet informative and useful. Currently, there are more than 850 subscribers from all over the world. Subscribers include hospital employees from major urban centers and obscure remote locales, university researchers, botanists and the employees of commercial laboratories, government agencies, veterinary facilities and a wide variety of commercial industrial ventures. WHO RUNS HISTONET? The list is run by Linda R. Margraf, M.D. and Herb K. Hagler, Ph.D. using hardware and software owned by the University of Texas Southwestern Medical School, Department of Pathology in Dallas, Texas. If you have any questions or problems with Histonet please contact Linda Margraf at LMargraf@childmed.dallas.tx.us. HOW DOES THE LIST WORK? This server, unlike many systems, uses ONLY ONE ADDRESS to send commands to the computer and to post messages. The server will recognize commands sent in the SUBJECT line of the message and only when they are spelled exactly as listed below. Anything not identified as a command will be circulated to EVERYONE on the list. The following is a list of commands the server recognizes: subscribe Your address will be added to the list of subscribers. You will then be able to send messages to this list that will be forwarded to all other list subscribers. You will begin to receive all messages sent to the list by other subscribers. subscribe digest Your address will be added to the list of subscribers who receive a digest instead of each forwarded message. A digest is a compilation of all the messages received in a 24 hour period. It is sent to the digest subscribers every night after midnight. Digest subscribers can post and respond to messages the same as "real-time" subscribers. digests A list of available digests will be returned to you. Histonet stores old messages as daily digests for approximately three months. To read previous messages, copy the list of available digests, mark the dates of interest and return it to the server. unsubscribe Your address will be removed from the list of subscribers. You will no longer be able to send messages to the members of the list. help A list of the commands recognized by the server will be returned to you. WHAT ARE THE RULES? You may post any questions you wish pertaining to histology, pathology, in-situ hybridization, immunohistochemistry etc. Equipment and reagent evaluations, laboratory management issues, government regulations, and job opportunities are all appropriate topics. The University asks that we restrict the use of its hardware and software to business purposes only (occasional jokes do slip through but PLEASE use restraint). Vendors and those with commercial interests in histology products are welcome contributors however, we ask that blatant advertisements be avoided at all times. It is fine to refer to product that your company produces if it is pertinent to a topic being discussed on the list. Unsolicited advertisements are poorly tolerated by the members and you will likely receive a number of negative comments if you overstep the boundaries. Please contact Linda Margraf at LMargraf@childmed.dallas.tx.us if you are not sure about the appropriateness if a message you wish to post. BASIC HISTONET "NETIQUETTE" It is most helpful to the list members if you post your responses to queries to everyone on the list and not just as a personal reply to the person asking the question. That way duplicate messages are minimized and we all learn from each other's comments. Likewise, if you post a question and get a number of responses back directly to you, it is helpful to everyone if you could send out a summary of the replies you got to Histonet. Please avoid abbreviations unless they are explained in your message. For example: immunohistochemistry (IHC). This list circulates to a wide variety of individuals and what seems obvious to you may have no meaning on the other side of the world. Please sign your letter and include your institution or affiliation and location. Not all email systems have headers which identify the sender. Do use the subject line to indicate the topic of your message. DON'T USE ONLY CAPITAL LETTERS -it is considered shouting. Please send questions and problems about the list directly to Linda Margraf at LMargraf@childmed.dallas.tx.us and don't circulate them to the >850 subscribers on the list. Be careful when sending commands to the server to put the command in the SUBJECT LINE and spell it correctly. Please do not send images as attachments with your message. We can now post images at our web site (http://pathcuri1.swmed.edu). To have an image posted send it to Herb Hagler at herb.hagler@email.swmed.edu. es From RBARNHART <@t> summithealth.org Fri Oct 15 11:36:44 2004 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Clo test Message-ID: I am trying to write a procedure for the clo test but I have misplaced in instruction packet. Can anyone share a procedure? Thanks Becky From cbrayton <@t> bcm.tmc.edu Fri Oct 15 12:10:00 2004 From: cbrayton <@t> bcm.tmc.edu (BCM HUMAN RESOURCES) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Re: Histonet Digest, Vol 11, Issue 20 Message-ID: <200410151710.MAA00396@morpheus.corp.bcm.tmc.edu> This user is no longer with Baylor College of Medicine. Thanks, Baylor College of Medicine From pruegg <@t> ihctech.net Fri Oct 15 13:15:27 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] silver stain with plastic embedded tendons In-Reply-To: Message-ID: <001201c4b2e2$f5e0f410$83020a0a@IHCTech> You might have a hard time keeping a section that thick on a slide. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Leclerc Jocelyne Sent: Friday, October 15, 2004 4:39 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] silver stain with plastic embedded tendons Hi, has anyone experience with silver stain on thick slide of plastic embedded horse tendon which are about 150 micrometer? We want to stain with Bodian and Rager. The tendons are embedded in MMA and the fixation was with formalin and glutaraldehyd. Thank you for your help. Jocelyne Leclerc VetSuisse Fakult?t Veterin?r-Anatomisches Institut Winterthurerstr. 260 8057 Z?rich _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MAUGER <@t> email.chop.edu Fri Oct 15 14:45:05 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] IHC withquantum dots Message-ID: Hi group, Can anyone help me with suggestions-I am working up the monoclonal Ab = (H2A10) to cat scratch disease. Ab from Novus Biologicus. I have a = purchased control from Newcomer Supply. I am using permanent red chromagen = from Dako., with alk phos link also from Dako. I am getting a nice = specific staining of what looks like specs vs tiny rods or spirochetes, = but the control tissue does , by Warthin Starry, look much more spirochete-= ish. Can I trust this result? I have also stained several suspected + = cases and see the same dust like + staining,but I am afraid it is teeny = bits of precipitate.=20 Does this ring a bell? Please help. Thanks, Jo >>> "John Kiernan" 10/13/04 09:57PM >>> Quantum dots are nanocrystals of a semiconductor (exact chemical identity seems to be trade secret). They may have a future in a new generation of tiny computers. Quantum dots fluoresce in a wide range of colours=20 and can be conjugated to antibodies, avidin, phalloidin etc. You can buy conjugated reagents or kits to conjugate your own. The Quantum Dot Corporation's web site http://www.qdots.com/live/index.asp=20 shows some pretty pictures of labelled cells. There is a recent paper about quantum dots in the J. Histochem. Cytochem. Here's the bibliographic information and abstract, from http://www.jhc.org/cgi/content/abstract/52/1/13=20 If your institution subscribes to the journal you'll be able to get the full text on line. --------- Rozalia Nisman, Graham Dellaire, Ying Ren, Ren Li and David P. Bazett=96Jones (2004) Application of Quantum Dots as Probes for Correlative Fluorescence, Conventional, and Energy-filtered Transmission Electron Microscopy=20 J Histochem Cystochem 52:13=9618 Luminescent semiconductor quantum dots (QDs) are a new class of fluorescent label with wide-ranging applications for cell imaging. The electron density and elemental composition of these materials permit the extension of their use as probes in conventional electron microscopy (TEM) and energy-filtered TEM (EFTEM). Here we illustrate the feasibility of using streptavidin-conjugated QDs as TEM tags by labeling a nuclear protein on cell sections and obtaining correlative fluorescence and TEM data. We also show that QD probes can be employed in conjunction with immunogold for co-localization of proteins at the ultrastructural level. Furthermore, by obtaining cadmium elemental maps of CdSe/ZnS QDs distributed on a nuclear structure, we demonstrate the potential of QDs for co-localization of multiple proteins when used in combination with EFTEM.=20 --------- A Google search for quantum dots brings up thousands of hits. Searching for "quantum dots" and histochemistry and cytochemistry brought up 60 hits, at least half of them in English and relevant. John Kiernan London, Canada. ----------------------------------------------- Patsy Ruegg wrote: >=20 > What are "quantum dots"? >=20 > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu=20 > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anna > Elisse Beaudin > Sent: Wednesday, October 13, 2004 7:55 AM > To: histonet@lists.utsouthwestern.edu=20 > Subject: [Histonet] IHC withquantum dots >=20 > Hello, >=20 > Has anyone ever done immunohistochemistry on tissue sections using > primary (or secondary) antibodies conjugated to quantum dots? if so, do > you think it would be possible to do multiple labeling using different > colored quantum dots? thanks in advance for your help! >=20 > Anna Beaudin > Division of Nutritional Sciences > Cornell University ------------------------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu=20 http://lists.utsouthwestern.edu/mailman/listinfo/histonet=20 From Rcartun <@t> harthosp.org Fri Oct 15 15:45:57 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] IHC withquantum dots Message-ID: Have you tried staining a section of tonsil to see if you get the same pattern of staining. I have found "precipitation" to be a problem with alkaline phosphatase (PA) detection, although I am by no means an expert when it comes to AP IHC. I use HRP/AEC for most of my infectious disease studies including Bartonella henselae. Don't forget that you may not see the normal morphology of the bacterium with IHC. You are labeling epitopes on the surface of the organism; you are not staining the entire cell wall. I have found B. henselae to be one of the most difficult organisms to label (and interpret) with IHC. Good luck! Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Joanne Mauger" 10/15/04 03:45PM >>> Hi group, Can anyone help me with suggestions-I am working up the monoclonal Ab (H2A10) to cat scratch disease. Ab from Novus Biologicus. I have a purchased control from Newcomer Supply. I am using permanent red chromagen from Dako., with alk phos link also from Dako. I am getting a nice specific staining of what looks like specs vs tiny rods or spirochetes, but the control tissue does , by Warthin Starry, look much more spirochete-ish. Can I trust this result? I have also stained several suspected + cases and see the same dust like + staining,but I am afraid it is teeny bits of precipitate. Does this ring a bell? Please help. Thanks, Jo >>> "John Kiernan" 10/13/04 09:57PM >>> Quantum dots are nanocrystals of a semiconductor (exact chemical identity seems to be trade secret). They may have a future in a new generation of tiny computers. Quantum dots fluoresce in a wide range of colours and can be conjugated to antibodies, avidin, phalloidin etc. You can buy conjugated reagents or kits to conjugate your own. The Quantum Dot Corporation's web site http://www.qdots.com/live/index.asp shows some pretty pictures of labelled cells. There is a recent paper about quantum dots in the J. Histochem. Cytochem. Here's the bibliographic information and abstract, from http://www.jhc.org/cgi/content/abstract/52/1/13 If your institution subscribes to the journal you'll be able to get the full text on line. --------- Rozalia Nisman, Graham Dellaire, Ying Ren, Ren Li and David P. Bazett*Jones (2004) Application of Quantum Dots as Probes for Correlative Fluorescence, Conventional, and Energy-filtered Transmission Electron Microscopy J Histochem Cystochem 52:13*18 Luminescent semiconductor quantum dots (QDs) are a new class of fluorescent label with wide-ranging applications for cell imaging. The electron density and elemental composition of these materials permit the extension of their use as probes in conventional electron microscopy (TEM) and energy-filtered TEM (EFTEM). Here we illustrate the feasibility of using streptavidin-conjugated QDs as TEM tags by labeling a nuclear protein on cell sections and obtaining correlative fluorescence and TEM data. We also show that QD probes can be employed in conjunction with immunogold for co-localization of proteins at the ultrastructural level. Furthermore, by obtaining cadmium elemental maps of CdSe/ZnS QDs distributed on a nuclear structure, we demonstrate the potential of QDs for co-localization of multiple proteins when used in combination with EFTEM. --------- A Google search for quantum dots brings up thousands of hits. Searching for "quantum dots" and histochemistry and cytochemistry brought up 60 hits, at least half of them in English and relevant. John Kiernan London, Canada. ----------------------------------------------- Patsy Ruegg wrote: > > What are "quantum dots"? > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anna > Elisse Beaudin > Sent: Wednesday, October 13, 2004 7:55 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] IHC withquantum dots > > Hello, > > Has anyone ever done immunohistochemistry on tissue sections using > primary (or secondary) antibodies conjugated to quantum dots? if so, do > you think it would be possible to do multiple labeling using different > colored quantum dots? thanks in advance for your help! > > Anna Beaudin > Division of Nutritional Sciences > Cornell University ------------------------------------------------------- _______________________________________________ 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 From Linresearch <@t> aol.com Fri Oct 15 16:58:13 2004 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Abs on Xenograft Tissues Message-ID: Hello, I need to do BRDU, Tunnel & Caspase on Xenograft rat tumors(FFPE) Is there anything that I need to do differently than when I run the same Abs on regular rat tissues? Lin From malek.j <@t> ghc.org Fri Oct 15 19:11:28 2004 From: malek.j <@t> ghc.org (Malek, Jack M) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Bile Stain Control Message-ID: <63EB2FB8C50AD242898B76E2B2C7C12202121996@ROC2T9.ghc.org> Hi All, Need help with our bile stain controls. We've tried three different tissues (2 blocks with liver bile stasis and a block of gall bladder) but no luck. We've also tried new reagents (TCA and Ferric Cholride)... still no luck. Does anyone have a working control block or two that they can send so that I can trouble shoot this? Thanks in advance, Jack Malek Group Health Cooperative 125 - 16th Avenue, E. CSB-A Pathology Seattle, WA 98177 206-326-3251 From cbrayton <@t> bcm.tmc.edu Sat Oct 16 12:09:21 2004 From: cbrayton <@t> bcm.tmc.edu (BCM HUMAN RESOURCES) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Re: Histonet Digest, Vol 11, Issue 21 Message-ID: <200410161709.MAA21153@cuda.corp.bcm.tmc.edu> This user is no longer with Baylor College of Medicine. Thanks, Baylor College of Medicine From contact <@t> excaliburpathology.com Sat Oct 16 13:04:27 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Silver stain with plastic embedded tendons Message-ID: <20041016180427.18012.qmail@web50307.mail.yahoo.com> Are the tendons that thick or are your sections this thick? Unless your are using a confocal microscope you will have problems if the sections are that thick. Not to mention keeping them on the slide as the other person stated, unless you float stained the section and then put it on a slide. And IMHO a silver stain would cause to much autofluorescents. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From cbrayton <@t> bcm.tmc.edu Sun Oct 17 12:04:54 2004 From: cbrayton <@t> bcm.tmc.edu (BCM HUMAN RESOURCES) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Re: Histonet Digest, Vol 11, Issue 22 Message-ID: <200410171704.MAA27623@morpheus.corp.bcm.tmc.edu> This user is no longer with Baylor College of Medicine. Thanks, Baylor College of Medicine From c.m.vanderloos <@t> amc.uva.nl Sun Oct 17 13:20:49 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] RE: AP problem Message-ID: <11310311304d.11304d113103@amc.uva.nl> Dear Joanne, Usually, AP staining doesn't show up in a crystal-like precipitate, but I have seen it a couple of times. Never found out what was the cause of it. Be sure that after your IHC procedure and before the AP visualization, the sections are washed at with a Tris-HCl pH8.5 buffer. Phosphate ions from PBS will inhibit your AP reaction. You also may mix the Permanent Red components first and than press immediately through a syringe-filter to remove all bigger particles in the AP solution. Have you viewed your slides under a fluorescence microscope? A small amount of Permanent Red reaction product also gives a bright red fluorescence signal when using the rhodamine filter pack (green light). Good suggestion by Richard to use a tonsil as a negative control. But, how about running the good old control blanc slide without any primary antibody? Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From "Joanne Mauger" Date Fri, 15 Oct 2004 15:45:05 -0400 To histonet@lists.utsouthwestern.edu, jkiernan@uwo.ca Subject Re: [Histonet] IHC withquantum dots Hi group, Can anyone help me with suggestions-I am working up the monoclonal Ab (H2A10) to cat scratch disease. Ab from Novus Biologicus. I have a purchased control from Newcomer Supply. I am using permanent red chromagen from Dako., with alk phos link also from Dako. I am getting a nice specific staining of what looks like specs vs tiny rods or spirochetes, but the control tissue does , by Warthin Starry, look much more spirochete-ish. Can I trust this result? I have also stained several suspected + cases and see the same dust like + staining,but I am afraid it is teeny bits of precipitate. Does this ring a bell? Please help. Thanks, Jo ----- Original Message ----- >From "Richard Cartun" Date Fri, 15 Oct 2004 16:45:57 -0400 To ,, Subject Re: [Histonet] IHC withquantum dots Have you tried staining a section of tonsil to see if you get the same pattern of staining. I have found "precipitation" to be a problem with alkaline phosphatase (PA) detection, although I am by no means an expert when it comes to AP IHC. I use HRP/AEC for most of my infectious disease studies including Bartonella henselae. Don't forget that you may not see the normal morphology of the bacterium with IHC. You are labeling epitopes on the surface of the organism; you are not staining the entire cell wall. I have found B. henselae to be one of the most difficult organisms to label (and interpret) with IHC. Good luck! Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 From LESCHUKC <@t> trinity-health.org Mon Oct 18 07:22:35 2004 From: LESCHUKC <@t> trinity-health.org (Carmen Leschuk) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] thin prep storage racks Message-ID: Anyone interested thin prep storage racks? I have some that I would like to get rid of (at a very low price compared to normal). Please email me and let me know! thanks. Carmen Leschuk Carmen Leschuk, HT (ASCP) Supervisor, SJMO-Anatomic Pathology (248)858-6231 From julien_lambreydesouza <@t> uqar.qc.ca Mon Oct 18 07:46:40 2004 From: julien_lambreydesouza <@t> uqar.qc.ca (Julien Lambrey De Souza) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] How to re-stain old specimen? Message-ID: <005301c4b510$840f83d0$cd11d784@rcloutier06> Hello to all the histonet community, We are doing the clear and double staining technique by Dinkerkus and Uhler (1977). For those who are not familiar with this technique, it consists in staining specimen with Alcian blue to reveal cartilage and with Alizarin Red to reveal bone, on whole fish that have been bleached (in H2O2 + KOH to remove chromatophores) and digested in trypsin (to make the fish transparent). The fish are then passed through a gradient series of KOH-Glycerin gradients to 100% Glycerin for storage (a few crystals of thymol are added to avoid mould formation). The technique works wonderfully well. But here is the catch. The problem is that the red Alizarin stain tends to fade with time and therefore all observations are done within 5 days of staining. In my particular case here, I have 2 old specimen (more than a year old) that have lost their red coloration but I have to retrieve bone information from them and I have no way of getting equivalent specimen to redo the whole staining procedure. So I have to re-stain these 2 fishes, go backwards in the protocol towards the red staining. According to the original protocol, I stain my samples in a KOH 0.5% solution of Alizarin red for a day. Then to remove excess red, I go through the above mentioned gradient series of KOH-Gylcerin towards the 100% glycerin for storage. I was thinking of reversing the gradient series in order to go back to KOH0.5% and Alizarin red, but in what proportions and where do I start incorporating the red stain? This is what i'm figuring: step 1: 100% Glycerin step 2: 75% Glycerin + 25% KOH(0.5%) step 3: 50% glycerin + 50% Alizarin red KOH(0.5%) solution step 4: 25% Glycerin + 75% red solution step 5: 100% red solution. Then I would gradually go back up to Glycerin 100%. But I'm wondering if this would be to much KOH for the specimen. I only have one shot at this, these 2 fish are valuable. This is why I'm looking for input on the subject (and where better than the histonet to find this). Any thoughts are very welcome. Thanks, Julien Lambrey de Souza Biologie ?volutive, Universit? du Qu?bec ? Rimouski, (418) 723-1986 #1438 From ljb <@t> medicine.wisc.edu Mon Oct 18 09:05:03 2004 From: ljb <@t> medicine.wisc.edu (LaCinda Burchell) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] IHC withquantum dots Message-ID: Dear Baowei Peng, Can you tell us why we can't co-localize? Cindy B LaCinda Burchell, BA, AS, HT(ASCP) University of Wisconsin-Madison, Medical School Asthma and Allergy Research IHC Lab 600 Highland Ave. CSC K4/913 Madison, Wisconsin 53792 Phone: 608-262-3518 FAX: 608-263-3746 >>> Baowei Peng 10/14/2004 2:20:32 AM >>> We have a fellow in our lab who worked on QDs (quantum dots). So I have learned that most QDs can be excitated under wave length about between 400-600nm. It means that you can use one certain wave length to excitate several different QDs which are distinctive in emission wave length (thus different in color). But the drawback is that you can not use two or more QDs in order to see colocalization such as nucleus and membrane molecules of the same cell. Baowei Peng 1954 Huashan Road Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China From mlm11 <@t> cornell.edu Mon Oct 18 09:14:50 2004 From: mlm11 <@t> cornell.edu (Mary Lou Norman) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] GAG staining Message-ID: <5.2.1.1.2.20041018094739.02908700@postoffice9.mail.cornell.edu> Hello Histonet, I need some insight on GAG staining please. I've done different times, different pH's. I have explants grown in aragose at 0 weeks that are not suppose to stain but do, and real tissue that's suppose to stain red and turns out green. I started with Lillie's (3rd ed.) procedure. Does anyone have a tried, true, and repeatable procedure? Is it my water, is it my staining dishes? Can dry stain go bad? Wit's end and other things to do. Thank you very much for any help. It's bovine tissue. Mary Lou From Barry.R.Rittman <@t> uth.tmc.edu Mon Oct 18 09:40:57 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] How to re-stain old specimen? Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0010076FB@UTHEVS3.mail.uthouston.edu> Julien I have some alizarin red specimens that were prepared over 25 years ago and some 40 years ago and the alizarin and alcian blue staining is still as good as when they were first stained. The secret is I believe to ensure that all of the potassium hydroxide is removed by using several changes of glycerin. I am not sure if storing in a closed container or the dark may also be a factor in preventing staining. I would recommend that you go backwards by adding small amounts of distilled water and gently mixing, over time until you are back at the original concentration. Then repeat the staining with alizarin in potassium hydroxide. Hope that this helps. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Julien Lambrey De Souza Sent: Monday, October 18, 2004 7:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] How to re-stain old specimen? Hello to all the histonet community, We are doing the clear and double staining technique by Dinkerkus and Uhler (1977). For those who are not familiar with this technique, it consists in staining specimen with Alcian blue to reveal cartilage and with Alizarin Red to reveal bone, on whole fish that have been bleached (in H2O2 + KOH to remove chromatophores) and digested in trypsin (to make the fish transparent). The fish are then passed through a gradient series of KOH-Glycerin gradients to 100% Glycerin for storage (a few crystals of thymol are added to avoid mould formation). The technique works wonderfully well. But here is the catch. The problem is that the red Alizarin stain tends to fade with time and therefore all observations are done within 5 days of staining. In my particular case here, I have 2 old specimen (more than a year old) that have lost their red coloration but I have to retrieve bone information from them and I have no way of getting equivalent specimen to redo the whole staining procedure. So I have to re-stain these 2 fishes, go backwards in the protocol towards the red staining. According to the original protocol, I stain my samples in a KOH 0.5% solution of Alizarin red for a day. Then to remove excess red, I go through the above mentioned gradient series of KOH-Gylcerin towards the 100% glycerin for storage. I was thinking of reversing the gradient series in order to go back to KOH0.5% and Alizarin red, but in what proportions and where do I start incorporating the red stain? This is what i'm figuring: step 1: 100% Glycerin step 2: 75% Glycerin + 25% KOH(0.5%) step 3: 50% glycerin + 50% Alizarin red KOH(0.5%) solution step 4: 25% Glycerin + 75% red solution step 5: 100% red solution. Then I would gradually go back up to Glycerin 100%. But I'm wondering if this would be to much KOH for the specimen. I only have one shot at this, these 2 fish are valuable. This is why I'm looking for input on the subject (and where better than the histonet to find this). Any thoughts are very welcome. Thanks, Julien Lambrey de Souza Biologie ?volutive, Universit? du Qu?bec ? Rimouski, (418) 723-1986 #1438 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From scoop <@t> mail.nih.gov Mon Oct 18 10:42:34 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] bone decal Message-ID: Dear Histonetters, Does anyone know if treating bone in 1% HCl (0.12N) for one hour would be adequate to decal it? (I have to treat the bone in the HCl prior to decal anyway.) Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From scoop <@t> mail.nih.gov Mon Oct 18 10:53:37 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] oil red O stain on liver Message-ID: Dear Histonetters, I'm planning to do oil red O stain on mouse liver. I'm planning to use the protocol in J. Kiernan's book, which is very clear except for a few things: Do I need to perfuse the liver prior to freezing or can I just fix in formalin prior to staining. Is formalin the best fixative for this technique? (I assume that methanol or acetone might dissolve some of the lipids and therefore be a bad choice, but maybe there's a reason to use them). If I need to perfuse the mice, do I just perfuse with PBS or should I also perfusion fix the tissue and with what fixative? Is it important to cryoprotect with sucrose before freezing (I've never done frozen sections on liver before, I get adequate results on brain without cryoprotecting). Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From kmerriam2003 <@t> yahoo.com Mon Oct 18 11:23:28 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] GAG staining In-Reply-To: <5.2.1.1.2.20041018094739.02908700@postoffice9.mail.cornell.edu> Message-ID: <20041018162328.15516.qmail@web52506.mail.yahoo.com> Mary Lou, At my last job, we did Alcian Blue stain for GAGs. You will need to check the textbooks to determine which pH to use, based on the type of GAGs you will be staining for. We actually fixed our tissues in methacarn, which yielded the best results for our particular application. I think we were using the Alcian Blue, pH 1.0; I'm pretty sure that this solution does go bad over time, and it needed to be made once per month or so. Alcian Blue is difficult to dissolve, so if you can buy the solution already made up, that would be the easiest thing to do. Hope this helps, Kim Merriam Novartis Cambridge, MA Mary Lou Norman wrote: Hello Histonet, I need some insight on GAG staining please. I've done different times, different pH's. I have explants grown in aragose at 0 weeks that are not suppose to stain but do, and real tissue that's suppose to stain red and turns out green. I started with Lillie's (3rd ed.) procedure. Does anyone have a tried, true, and repeatable procedure? Is it my water, is it my staining dishes? Can dry stain go bad? Wit's end and other things to do. Thank you very much for any help. It's bovine tissue. Mary Lou _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From n.galletly <@t> imperial.ac.uk Mon Oct 18 11:30:28 2004 From: n.galletly <@t> imperial.ac.uk (Galletly, Neil P) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Fresh tissue sectioning and safety Message-ID: I plan to cut 200 micron sections of fresh human tissue (mainly colon, esophagus and cervix) using a Krumdieck Tissue Slicer as part of a research project. Our college safety officer has raised concerns about the possibility of tissue aerosol generation during cutting the tissue, which would have implications regarding possible spread of viral infections such as HIV and Hep B/C etc. I'd be grateful for people's views as to whether this is likely to be a real risk and if so what precautions people are taking to avoid it. I could use a Vibratome instead but I understand this is much more of a hassle - and presumably the risk of aerosol formation would still exist? Many thanks Neil Galletly Clinical Research Fellow Department of Histopathology Imperial College London Hammersmith Campus London UK From MRSmith <@t> TarrantCounty.com Mon Oct 18 11:35:16 2004 From: MRSmith <@t> TarrantCounty.com (Michael R. Smith) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Information on immunohistochemistry stain on paraffin embedded tissue for troponin I Message-ID: I am looking for anyone who has done an immunohistochemistry stain for troponin I on paraffin embedded tissue. One of the pathologists at the Medical Examiners office wants to do research in this area. Specifically, where we can get the antibody from, the protocol and any other useful information. Thank you for your help in this area. Mike Smith HT (ASCP) Tarrant County Medical Examiner Office Fort Worth, Texas 76104 From cbrayton <@t> bcm.tmc.edu Mon Oct 18 12:03:35 2004 From: cbrayton <@t> bcm.tmc.edu (BCM HUMAN RESOURCES) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] Re: Histonet Digest, Vol 11, Issue 23 Message-ID: <200410181703.MAA22223@morpheus.corp.bcm.tmc.edu> This user is no longer with Baylor College of Medicine. Thanks, Baylor College of Medicine From vazquezr <@t> ohsu.edu Mon Oct 18 12:11:11 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:10 2005 Subject: [Histonet] bone decal Message-ID: Is this for a frozen section or to process...and is the bone small or large? Robyn OHSU Portland, Or >>> Sharon Cooperman 10/18/2004 8:42:34 AM >>> Dear Histonetters, Does anyone know if treating bone in 1% HCl (0.12N) for one hour would be adequate to decal it? (I have to treat the bone in the HCl prior to decal anyway.) Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Mon Oct 18 12:27:09 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] bone decal In-Reply-To: Message-ID: <002801c4b537$b2ad5830$83020a0a@IHCTech> It depends on how thick and dense the bone is, 1%HCL is not very strong but if it is a small bone biopsy it might work, we need more info. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sharon Cooperman Sent: Monday, October 18, 2004 8:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] bone decal Dear Histonetters, Does anyone know if treating bone in 1% HCl (0.12N) for one hour would be adequate to decal it? (I have to treat the bone in the HCl prior to decal anyway.) Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From scoop <@t> mail.nih.gov Mon Oct 18 12:39:33 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] re: bone decal - clarification Message-ID: Hi Histonetters, Thanks to everyone who has answered my bone decal question so far - I'm writing to add a clarification - the bone is mouse femur (pretty small and thin) about one year old. I'm treating it in HCl because I'm putting it in Perl's solution with 4% paraformaldehyde first before anything else to preserve the iron, which I find leaches out during fixation. It works well, probably because formalin is more stable in acid solution. I'm just wondering if maybe I don' have to go through the decal afterwards. I guess if the one hour in 1% HCL isn't enough for decal then I should use a regular decal step rather than continue in 1% HCl because that would better preserve antigens for subsequent IHC. Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From nhiggs <@t> bioforminc.com Mon Oct 18 13:02:50 2004 From: nhiggs <@t> bioforminc.com (Nicole Higgs) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Gomori & Sorensen's Buffer Message-ID: <033BF6EFEC28AE47AF707245117111EB0E7A4F@wi-dc.bioforminc.com> Hello All, I am a newcomer to the Histonet listserve, and I do not work in the histology field, but I thought someone in this group might be able to help me. I am wondering if anyone has a recipe or reference for Gomori's Buffer that they could share with me? I am not familiar with this buffer and I have been having a difficult time finding it. I am thinking it's a component to a histological stain?? I have also been looking for a recipe for Sorensen's buffer. I have found Sorensen's Phosphate buffer in the archives of this list serve, but I was also wondering if Sorensen's Citrate buffer was also familiar to anyone? Are these buffers just standard PBS and citrate buffer's? I would like to use these buffers to compare how different calcium hydroxylapatite (a bone substitute used in orthopedics and augmentation) compounds dissolves over time. Thanks for your help, Nicole S. Higgs R&D Technician BioForm Medical, Inc. 4133 Courtney Road, # 10 Franksville, WI 53126 Phone: 262-835-3315 Fax: 262-835-9311 E-Mail: nhiggs@bioforminc.com Web: http://www.bioforminc.com/ From cwscouten <@t> myneurolab.com Mon Oct 18 16:40:01 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Fresh tissue sectioning and safety Message-ID: I doubt that anyone has measured aresol formation during use of a tissue chopper or a Vibratome. The existance of such an aerosol formation effect is hypothetical, but has not been ruled out. We offer both a McIllwain Tissue chopper, and the Vibratome(TM). If I may speculate, a Chopper impacts the tissue and drives through. There is much more violence than a Vibratome would create, and could be splashes and spray. Cells might be squeezed, and squirt. A Vibratome saws its way through the tissue. I can not see any aresol formation likely with the Vibratome, but, again, it has never been measured. As a former Vibratome user and now a representative, I do not think you would find it any more a "hassle" than the choppers. The instrument comes with a supply of instant setting cyanoacrylate glue. Just glue the tissue to a pedastal, put it in the Vibratome, and cut your sections. The Vibratome can cut 50 micron fresh sections, much thinner than a chopper, but 200 would be easy work. Just cut it. Drain the fluid out the tap in the back, and bleach or other stong disinfectant can be run through, then rinse. Do you have access to a Vibratome to have someone show you how it works? Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Galletly, Neil P Sent: Monday, October 18, 2004 11:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Fresh tissue sectioning and safety I plan to cut 200 micron sections of fresh human tissue (mainly colon, esophagus and cervix) using a Krumdieck Tissue Slicer as part of a research project. Our college safety officer has raised concerns about the possibility of tissue aerosol generation during cutting the tissue, which would have implications regarding possible spread of viral infections such as HIV and Hep B/C etc. I'd be grateful for people's views as to whether this is likely to be a real risk and if so what precautions people are taking to avoid it. I could use a Vibratome instead but I understand this is much more of a hassle - and presumably the risk of aerosol formation would still exist? Many thanks Neil Galletly Clinical Research Fellow Department of Histopathology Imperial College London Hammersmith Campus London UK _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From conniegrubaugh <@t> hotmail.com Mon Oct 18 18:54:38 2004 From: conniegrubaugh <@t> hotmail.com (connie grubaugh) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Nevada Society Of Histechnology Message-ID: Just a reminder that the NVSH will be meeting Oct. 29th and 30th. Fridays meeting begins at 6pm and Sat. at 9 am. The weather is always great in October in Las Vegas and just the place to spend Halloween... Please let me know if you need more information and I will fax it to you. Hope to see you in Vegas.................... Connie G. _________________________________________________________________ [1]Rock, jazz, country, soul & more. Find the music you love on MSN Music! References 1. http://g.msn.com/8HMBENUS/2740??PS=47575 From pengbw <@t> sjtu.edu.cn Mon Oct 18 20:48:58 2004 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] IHC withquantum dots Message-ID: <20041019014858.5798110CD52D@sjtu.edu.cn> Dear LaCinda I draw the conclusion based on my experience. I have no idea of the mechanism or explain to this phenomena. What I had done was using QDs as a tracer which was thought being taken up by macrophages and/or DCs. Then I made frozen sections of the draining lymph node (LN) which drain the tissue where QDs were applied. I used DAPI to stain the nucleus of cell and after that use UV to excitated both DAPI and QDs. What I had seen was that blue(nucleus) and yellow (QDs) were distinctive. The two color never overlap as I had thought. I\\\'m sure there should some cells in the drain LN had taken up QDs. I think since both blue(nucleus) and yellow (QDs) were excitated by UV at the same time, they may interfere each other and only one color(wavelength) emisson. Hope this make can help! Baowei Peng ----- Original Message ----- From: "LaCinda Burchell" To: , Cc: Subject: Re: [Histonet] IHC withquantum dots Dear Baowei Peng, Can you tell us why we can\'t co-localize? Cindy B LaCinda Burchell, BA, AS, HT(ASCP) University of Wisconsin-Madison, Medical School Asthma and Allergy Research IHC Lab 600 Highland Ave. CSC K4/913 Madison, Wisconsin 53792 Phone: 608-262-3518 FAX: 608-263-3746 >>> Baowei Peng 10/14/2004 2:20:32 AM >>> We have a fellow in our lab who worked on QDs (quantum dots). So I have learned that most QDs can be excitated under wave length about between 400-600nm. It means that you can use one certain wave length to excitate several different QDs which are distinctive in emission wave length (thus different in color). But the drawback is that you can not use two or more QDs in order to see colocalization such as nucleus and membrane molecules of the same cell. Baowei Peng 1954 Huashan Road Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China From Jackie.O'Connor <@t> abbott.com Tue Oct 19 07:01:25 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Dog antibody search Message-ID: Hi guys - Can anyone steer me towards a supplier for Caspase-3, MPM-2, and Ki-67 primary antibodies that will work for IHC on FFPE Dog?? Thanks much! Jackie O' From lfit4832 <@t> mail.usyd.edu.au Tue Oct 19 07:34:36 2004 From: lfit4832 <@t> mail.usyd.edu.au (lfit4832@mail.usyd.edu.au) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Dog antibody search In-Reply-To: References: Message-ID: <1098189276.417509dc0cf94@www-mail.usyd.edu.au> Hi Jackie, The only Ki-67 antibody successfully used in canine tissue (in the literature I have read anyway) is the Immunotech MIB-1 clone. Immunotech antibodies are now distributed by Beckman Coulter, and are indicated by (IM) in front of the catalogue number - think the catalogue number for the Ki67 (MIB-1) monoclonal is IM1316. I have 2ml of it in the fridge and I am planning to run it on a bunch of canien tumours over the summer. Here are a few references: Geraldes, M., F. Gartner, et al. (2000). "Immunohistochemical study of hormonal receptors and cell proliferation in normal canine mammary glands and spontaneous mammary tumours." Veterinary Record 146(14): 403-6. Laprie, C., J. Abadie, et al. (2001). "MIB-1 immunoreactivity correlates with biologic behaviour in canine cutaneous melanoma." Veterinary Dermatology 12(3): 139-47. Madewell, B. R. (2001). "Cellular proliferation in tumors: a review of methods, interpretation, and clinical applications." Journal of Veterinary Internal Medicine 15(4): 334-40. Millanta, F., F. Fratini, et al. (2002). "Proliferation activity in oral and cutaneous canine melanocytic tumours: correlation with histological parameters, location, and clinical behaviour." Research in Veterinary Science 73(1): 45-51. Perez Alenza, M. D., L. Pena, et al. (2000). "Factors influencing the incidence and prognosis of canine mammary tumours.[see comment]." Journal of Small Animal Practice 41(7): 287-91. Phillips, B. S. (2000). "Apoptotic and proliferation indexes in canine lymphoma." Journal of Veterinary Diagnostic Investigation 12(2): 111-117. Roels, S., K. Tilmant, et al. (1999). "PCNA and Ki67 proliferation markers as criteria for prediction of clinical behaviour of melanocytic tumours in cats and dogs." Journal of Comparative Pathology 121(1): 13-24. Zacchetti, A., E. van Garderen, et al. (2003). "Validation of the use of proliferation markers in canine neoplastic and non-neoplastic tissues: comparison of KI-67 and proliferating cell nuclear antigen (PCNA) expression versus in vivo bromodeoxyuridine labelling by immunohistochemistry." APMIS 111(3): 430-8. Cheers, Louise Quoting Jackie.O'Connor@abbott.com: > Hi guys - > Can anyone steer me towards a supplier for Caspase-3, MPM-2, and Ki-67 > primary antibodies that will work for IHC on FFPE Dog?? > > Thanks much! > > Jackie O' > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ---------------------------------------------------------------- This message was sent using IMP, the Internet Messaging Program. From Jill.Cox <@t> providence.org Tue Oct 19 09:31:09 2004 From: Jill.Cox <@t> providence.org (Cox, Jill) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Nevada Society Of Histechnology Message-ID: <05424658AAA6D441BBAFD9D9D98780B7044F0CC4@lcmmsg02.ca.providence.org> Looking forward to it!!! See you there. Jill -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of connie grubaugh Sent: Monday, October 18, 2004 4:55 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Nevada Society Of Histechnology Just a reminder that the NVSH will be meeting Oct. 29th and 30th. Fridays meeting begins at 6pm and Sat. at 9 am. The weather is always great in October in Las Vegas and just the place to spend Halloween... Please let me know if you need more information and I will fax it to you. Hope to see you in Vegas.................... Connie G. _________________________________________________________________ [1]Rock, jazz, country, soul & more. Find the music you love on MSN Music! References 1. http://g.msn.com/8HMBENUS/2740??PS=47575 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet DISCLAIMER: This message is intended for the sole use of the addressee, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If you are not the addressee you are hereby notified that you may not use, copy, disclose, or distribute to anyone the message or any information contained in the message. If you have received this message in error, please immediately advise the sender by reply email and delete this message. From jmahoney <@t> alegent.org Tue Oct 19 09:52:31 2004 From: jmahoney <@t> alegent.org (Janice A Mahoney) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Acid cleaning slides Message-ID: Hello, I would like to know if anyone has had problems with attempting to acid clean plus slides. Does this process affect the adhesion of cells to the slide. We have some background staining on our HPV thin prep slides and are trying to eliminate the background. Thanks, Jan Mahoney Omaha NE From dobbin <@t> upei.ca Tue Oct 19 10:12:33 2004 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Acid cleaning slides In-Reply-To: Message-ID: <417504B0.12762.AC86AE@localhost> Hi Janice, I certainly may be corrected on this, but I don't believe your "Plus" slides will be "Plus" slides anymore if you acid wash them. I would think that an acid wash would remove the positive charge from the slides. A few years ago, when I was coating/charging my own slides, I would acid wash and alcohol dry regular glass slides prior to treatment with Poly-L Lysine or Silane (APES). The acid wash helped remove any oily film that may have accumulated on the slides in storage, that might have interfered with proper coating of the Poly-L or APES. Greg Date sent: Tue, 19 Oct 2004 09:52:31 -0500 From: "Janice A Mahoney" To: Subject: [Histonet] Acid cleaning slides > Hello, > I would like to know if anyone has had problems with attempting to acid clean plus slides. Does this process affect the adhesion of cells to the slide. We have some background staining on our HPV thin prep slides and are trying to eliminate the background. > Thanks, > Jan Mahoney > Omaha NE > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Pathology Lab Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Happiness is a journey, not a destination. From cbrayton <@t> bcm.tmc.edu Tue Oct 19 12:07:36 2004 From: cbrayton <@t> bcm.tmc.edu (BCM HUMAN RESOURCES) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Re: Histonet Digest, Vol 11, Issue 24 Message-ID: <200410191707.MAA26627@morpheus.corp.bcm.tmc.edu> This user is no longer with Baylor College of Medicine. Thanks, Baylor College of Medicine From pruegg <@t> ihctech.net Tue Oct 19 12:41:47 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] re: bone decal - clarification In-Reply-To: Message-ID: <001601c4b602$e817cc00$83020a0a@IHCTech> Sharon, Decal can leach out the iron also. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sharon Cooperman Sent: Monday, October 18, 2004 10:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] re: bone decal - clarification Hi Histonetters, Thanks to everyone who has answered my bone decal question so far - I'm writing to add a clarification - the bone is mouse femur (pretty small and thin) about one year old. I'm treating it in HCl because I'm putting it in Perl's solution with 4% paraformaldehyde first before anything else to preserve the iron, which I find leaches out during fixation. It works well, probably because formalin is more stable in acid solution. I'm just wondering if maybe I don' have to go through the decal afterwards. I guess if the one hour in 1% HCL isn't enough for decal then I should use a regular decal step rather than continue in 1% HCl because that would better preserve antigens for subsequent IHC. Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From caroline.stott <@t> anatomy.otago.ac.nz Tue Oct 19 18:15:44 2004 From: caroline.stott <@t> anatomy.otago.ac.nz (Caroline Stott) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Chromic acid substitute Message-ID: <5.2.1.1.0.20041020121335.0204eec0@anatomy.otago.ac.nz> Hi all, Just when I thought I had rid the lab of all chromic acid, the boss wants to use it again. What are other labs using as a safer alternative to chromic acid for washing slides? Can you please give me some recipes? Thanks Caroline Caroline Stott Histology Service Unit University of Otago PO Box 913 Dunedin, New Zealand Ph (03) 479 7152 Fax (03) 479 7136 From WWmn916 <@t> aol.com Tue Oct 19 23:56:23 2004 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Eosin, phloxine and OG? Message-ID: <15a.41621350.2ea749f7@aol.com> Hello everyone, Say, is anyone familiar with an eosin/phloxine that mixes into it some OG (for H+E staining)? The conversation came up recently to "amp" the three shades of red in H+E staining. It was the first time I'd ever heard of adding Orange G into eosin/phloxing solution. Are there any labs doing this? Thanks:-) Deb From bills <@t> icpmr.wsahs.nsw.gov.au Wed Oct 20 00:12:00 2004 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Eosin, phloxine and OG? In-Reply-To: <15a.41621350.2ea749f7@wsahs.nsw.gov.au> Message-ID: <000001c4b663$55293d00$83a7080a@wsahs.nsw.gov.au> I haven't heard of Orange G being used before. But we use an aqueous solution of 1% eosin yellowish and 1% erythrosin B in 0.25% Sodium Hydrogen Carbonate and 2% Magnesium sulphate. We have used this for more than 40 years with great results all the time. Depending on the pH of your local water it may require a wash for anywhere between 10 secs to 1 min. Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of WWmn916@aol.com Sent: Wednesday, 20 October 2004 2:56 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Eosin, phloxine and OG? Hello everyone, Say, is anyone familiar with an eosin/phloxine that mixes into it some OG (for H+E staining)? The conversation came up recently to "amp" the three shades of red in H+E staining. It was the first time I'd ever heard of adding Orange G into eosin/phloxing solution. Are there any labs doing this? Thanks:-) Deb _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. From jkiernan <@t> uwo.ca Wed Oct 20 00:26:00 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Eosin, phloxine and OG? References: <15a.41621350.2ea749f7@aol.com> Message-ID: <4175F6E8.7DCD7121@uwo.ca> If you look at your H&E slides between staining and clearing & coverslipping you will find that eosin Y (CI 45380) can impart more than 3 colours to RBC, smooth muscle cytoplasm, other cytoplasms and collagen. Adding phloxin or orange G to an eosin Y counterstain may extend the spectrum, but don't expect clear-cut results from slides stained by a machine. The H&E method needs human judgement and adjustments before applying a coverslip. For this reason and others H&E is not a good "routine" staining method. It's all there in the peer-reviewed literature and in most histotechnology textbooks less than 25 years old. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ > (for H+E staining)? > The conversation came up recently to "amp" the three shades of red in H+E > staining. It was the first time I'd ever heard of adding Orange G into > eosin/phloxing solution. Are there any labs doing this? > > Thanks:-) > Deb > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rentonlf <@t> bru.wits.ac.za Wed Oct 20 06:41:02 2004 From: rentonlf <@t> bru.wits.ac.za (renton louise mrs) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] processing schedule Message-ID: <1098272462.8e42b040rentonlf@bru.wits.ac.za> Hello, could someone please advise me as to how to adjust a processing schedule for a "dip & dunk" processor to one in which heat and vacuum is an option? I am processing non decalcified baboon bone - specifically full thickness jaw (no teeth, thank goodness) and skull? The thickness obviously varies from 0.5 - 1cm and the area is about 2cm square. In the didp & dunk we generally left the specimen for 24hrs per beaker, but I am sure these times could be reduced. Thank you in advance & best regards Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? From cnagamin <@t> MIT.EDU Wed Oct 20 07:15:06 2004 From: cnagamin <@t> MIT.EDU (Claude M Nagamine) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] intestines fixation Message-ID: <1098274506.417656ca90a6a@webmail.mit.edu> I'm looking for tumors in the small and large intestines of mice. I've been inflating the intestines with formalin immediately after removal to prevent autolysis. It also stretches the intestines making them easier to cut open and, if things work well, flushes the lumen. However, there are often "tight" constricted areas that do not inflate or flush. I was thinking that things would be simple if the intestines were flaccid in the first place. Does anyone have a technique to relax the intestines prior to flushing with formalin? Claude Nagamine MIT, Div. Comp. Medicine Cambridge, MA From Michael.Rice <@t> holy-cross.com Wed Oct 20 07:40:21 2004 From: Michael.Rice <@t> holy-cross.com (Rice, Michael) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] processing schedule Message-ID: <3BC92F29BE821745AB15E04C98EE028D69361B@HCH2KMAIL.holy-cross.com> arent baboons an endangered species? -----Original Message----- From: renton louise mrs [mailto:rentonlf@bru.wits.ac.za] Sent: Wednesday, October 20, 2004 7:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] processing schedule Hello, could someone please advise me as to how to adjust a processing schedule for a "dip & dunk" processor to one in which heat and vacuum is an option? I am processing non decalcified baboon bone - specifically full thickness jaw (no teeth, thank goodness) and skull? The thickness obviously varies from 0.5 - 1cm and the area is about 2cm square. In the didp & dunk we generally left the specimen for 24hrs per beaker, but I am sure these times could be reduced. Thank you in advance & best regards Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- From MAUGER <@t> email.chop.edu Wed Oct 20 07:47:06 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] RE: AP problem Message-ID: Thanks all for the cat scratch advice. I am trying it with HRP- AEC. Will report result. Jo >>> "C.M. van der Loos" 10/17/04 02:20PM >>> Dear Joanne, Usually, AP staining doesn't show up in a crystal-like precipitate, but I have seen it a couple of times. Never found out what was the cause of it. Be sure that after your IHC procedure and before the AP visualization, the sections are washed at with a Tris-HCl pH8.5 buffer. Phosphate ions from PBS will inhibit your AP reaction. You also may mix the Permanent Red components first and than press immediately through a syringe-filter to remove all bigger particles in the AP solution. Have you viewed your slides under a fluorescence microscope? A small amount of Permanent Red reaction product also gives a bright red fluorescence signal when using the rhodamine filter pack (green light). Good suggestion by Richard to use a tonsil as a negative control. But, how about running the good old control blanc slide without any primary antibody? Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From "Joanne Mauger" Date Fri, 15 Oct 2004 15:45:05 -0400 To histonet@lists.utsouthwestern.edu, jkiernan@uwo.ca Subject Re: [Histonet] IHC withquantum dots Hi group, Can anyone help me with suggestions-I am working up the monoclonal Ab (H2A10) to cat scratch disease. Ab from Novus Biologicus. I have a purchased control from Newcomer Supply. I am using permanent red chromagen from Dako., with alk phos link also from Dako. I am getting a nice specific staining of what looks like specs vs tiny rods or spirochetes, but the control tissue does , by Warthin Starry, look much more spirochete-ish. Can I trust this result? I have also stained several suspected + cases and see the same dust like + staining,but I am afraid it is teeny bits of precipitate. Does this ring a bell? Please help. Thanks, Jo ----- Original Message ----- >From "Richard Cartun" Date Fri, 15 Oct 2004 16:45:57 -0400 To ,, Subject Re: [Histonet] IHC withquantum dots Have you tried staining a section of tonsil to see if you get the same pattern of staining. I have found "precipitation" to be a problem with alkaline phosphatase (PA) detection, although I am by no means an expert when it comes to AP IHC. I use HRP/AEC for most of my infectious disease studies including Bartonella henselae. Don't forget that you may not see the normal morphology of the bacterium with IHC. You are labeling epitopes on the surface of the organism; you are not staining the entire cell wall. I have found B. henselae to be one of the most difficult organisms to label (and interpret) with IHC. Good luck! Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 From Jackie.O'Connor <@t> abbott.com Wed Oct 20 07:53:53 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] processing schedule Message-ID: You wouldn't think so if you took a look around my office. "Rice, Michael" Sent by: histonet-bounces@lists.utsouthwestern.edu 10/20/2004 07:40 AM To: "renton louise mrs" , cc: Subject: RE: [Histonet] processing schedule arent baboons an endangered species? -----Original Message----- From: renton louise mrs [mailto:rentonlf@bru.wits.ac.za] Sent: Wednesday, October 20, 2004 7:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] processing schedule Hello, could someone please advise me as to how to adjust a processing schedule for a "dip & dunk" processor to one in which heat and vacuum is an option? I am processing non decalcified baboon bone - specifically full thickness jaw (no teeth, thank goodness) and skull? The thickness obviously varies from 0.5 - 1cm and the area is about 2cm square. In the didp & dunk we generally left the specimen for 24hrs per beaker, but I am sure these times could be reduced. Thank you in advance & best regards Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From David.Rushworth <@t> mail.bhrv.nwest.nhs.uk Wed Oct 20 07:58:39 2004 From: David.Rushworth <@t> mail.bhrv.nwest.nhs.uk (David Rushworth) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] blood-borne infections and the cryostat.#[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D0103F8A6@BHRV_NT_11> Our SOP states that frozen sections on tissue from patients with blood-borne infections etc. should not be done. This has been custom and practice for as long as I've worked here. After various phone calls to surrounding labs this appears to be their custom as well. After looking through the current HSE Safe working and prevention book 119 states that you decontaminate the cryostat if contaminated with infective material. My question, are there any new guidelines out there that I've missed? Or do I stick with refusing to do them. P.S. cryostat not in CL3 room. Hopefully some info. could help me out of a situation!! Thank in anticipation. Dave. From JWEEMS <@t> sjha.org Wed Oct 20 08:04:18 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] blood-borne infections and the cryostat.#[Scanned] Message-ID: <83AACDB0810528418AA106F9AE9B7F7E278517@sjhaexc02.sjha.org> How can you be sure that the material is not infective? Universal precautions should prevail. If you are CAP approved, they recommend weekly cleaning/disinfecting if the cryostat is used daily. http://www.cap.org/apps/docs/cap_today/q_and_a/qa_05_04.html This site has the info. Good luck! Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of David Rushworth Sent: Wednesday, October 20, 2004 8:59 AM To: histonet@pathology.swmed.edu Subject: [Histonet] blood-borne infections and the cryostat.#[Scanned] Our SOP states that frozen sections on tissue from patients with blood-borne infections etc. should not be done. This has been custom and practice for as long as I've worked here. After various phone calls to surrounding labs this appears to be their custom as well. After looking through the current HSE Safe working and prevention book 119 states that you decontaminate the cryostat if contaminated with infective material. My question, are there any new guidelines out there that I've missed? Or do I stick with refusing to do them. P.S. cryostat not in CL3 room. Hopefully some info. could help me out of a situation!! Thank in anticipation. Dave. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From JWEEMS <@t> sjha.org Wed Oct 20 08:05:18 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] RE: Controls for Immunos fixed in B-Plus Message-ID: <83AACDB0810528418AA106F9AE9B7F7E278518@sjhaexc02.sjha.org> I haven't heard from anyone! Let me ask again. Thanks, Joyce -----Original Message----- From: Weems, Joyce Sent: Friday, October 15, 2004 11:28 AM To: 'Histonet' Subject: Controls for Immunos fixed in B-Plus Hi Everyone, Do you use B-Plus fixed control tissue for immunos done on B-Plus fixed patient tissue? If so, how does it compare to formalin or B-5? Thanks in advance. j And happy weekend to everyone! j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. -------------- next part -------------- Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s Health System, Inc. From Barry.R.Rittman <@t> uth.tmc.edu Wed Oct 20 08:06:02 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] processing schedule Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0010077D1@UTHEVS3.mail.uthouston.edu> If so we would have a lot less politicians. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rice, Michael Sent: Wednesday, October 20, 2004 7:40 AM To: renton louise mrs; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] processing schedule arent baboons an endangered species? -----Original Message----- From: renton louise mrs [mailto:rentonlf@bru.wits.ac.za] Sent: Wednesday, October 20, 2004 7:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] processing schedule Hello, could someone please advise me as to how to adjust a processing schedule for a "dip & dunk" processor to one in which heat and vacuum is an option? I am processing non decalcified baboon bone - specifically full thickness jaw (no teeth, thank goodness) and skull? The thickness obviously varies from 0.5 - 1cm and the area is about 2cm square. In the didp & dunk we generally left the specimen for 24hrs per beaker, but I am sure these times could be reduced. Thank you in advance & best regards Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ajennings <@t> unmc.edu Wed Oct 20 08:10:35 2004 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] intestines fixation In-Reply-To: <1098274506.417656ca90a6a@webmail.mit.edu> Message-ID: Are you sure your "tight" constricted areas are not actually places where your "tumors" are obstructing the lumen? Do you get this same "tightness" in your controls? Is there any concern that inflating the intestines until they "stretch" may be distorting your morphology. Does your protocol allow for fasting before collection? I'm looking for tumors in the small and large intestines of mice. I've been inflating the intestines with formalin immediately after removal to prevent autolysis. It also stretches the intestines making them easier to cut open and, if things work well, flushes the lumen. However, there are often "tight" constricted areas that do not inflate or flush. I was thinking that things would be simple if the intestines were flaccid in the first place. Does anyone have a technique to relax the intestines prior to flushing with formalin? Claude Nagamine MIT, Div. Comp. Medicine Cambridge, MA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Wed Oct 20 11:36:35 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] intestines fixation In-Reply-To: <1098274506.417656ca90a6a@webmail.mit.edu> References: <1098274506.417656ca90a6a@webmail.mit.edu> Message-ID: <41769413.3090804@umdnj.edu> Hi Claude: Your problem may be that the intestines are attached to the posterior abdominal wall with a mesentary. Getting the intestine straight, without the normal curves and kinks, means you will have to cut the mesentary. Flushing with fixative is a good idea but keep the 'inflation' mild to avoid distortion. Another poster suggested that the constricted areas might be due to the tumors themselves, sounds like a possibility to me too. Unfixed intestine is pretty 'relaxed' already and if you want good fixation, fix the tissue immediately after death. Don't worry about making it more flaccid. Geoff Claude M Nagamine wrote: >I'm looking for tumors in the small and large intestines of mice. I've been >inflating the intestines with formalin immediately after removal to prevent >autolysis. It also stretches the intestines making them easier to cut open >and, if things work well, flushes the lumen. However, there are often "tight" >constricted areas that do not inflate or flush. I was thinking that >things would be simple if the intestines were flaccid in the first place. > >Does anyone have a technique to relax the intestines prior to flushing with >formalin? > >Claude Nagamine >MIT, Div. Comp. Medicine >Cambridge, MA > > > >_______________________________________________ >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; fax: -4029 mcauliff@umdnj.edu ********************************************** From Michael.Rice <@t> holy-cross.com Wed Oct 20 08:35:40 2004 From: Michael.Rice <@t> holy-cross.com (Rice, Michael) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] processing schedule Message-ID: <3BC92F29BE821745AB15E04C98EE028D69361C@HCH2KMAIL.holy-cross.com> Maybe we should use politicians for research instead of baboons -----Original Message----- From: Barry R Rittman [mailto:Barry.R.Rittman@uth.tmc.edu] Sent: Wednesday, October 20, 2004 9:06 AM To: histonet Subject: RE: [Histonet] processing schedule If so we would have a lot less politicians. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rice, Michael Sent: Wednesday, October 20, 2004 7:40 AM To: renton louise mrs; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] processing schedule arent baboons an endangered species? -----Original Message----- From: renton louise mrs [mailto:rentonlf@bru.wits.ac.za] Sent: Wednesday, October 20, 2004 7:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] processing schedule Hello, could someone please advise me as to how to adjust a processing schedule for a "dip & dunk" processor to one in which heat and vacuum is an option? I am processing non decalcified baboon bone - specifically full thickness jaw (no teeth, thank goodness) and skull? The thickness obviously varies from 0.5 - 1cm and the area is about 2cm square. In the didp & dunk we generally left the specimen for 24hrs per beaker, but I am sure these times could be reduced. Thank you in advance & best regards Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- _______________________________________________ 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 ---------------------- Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- From jcox90 <@t> yahoo.com Wed Oct 20 08:39:56 2004 From: jcox90 <@t> yahoo.com (Jill Cox) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] (no subject) Message-ID: <20041020133956.91384.qmail@web40913.mail.yahoo.com> Hello Netters, I want to thank you all again for all of the VERY useful tips on starting up my microtomy service. It’s up and running and I have my first client. The website is up also, www.precisionmicrotomy.com . I will be making a link for the email and a couple other changes. If anyone has any ideas or comments I would love to hear them. Jill Jill Cox HT (ASCP) --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From Jacqueline.Miller <@t> UTSouthwestern.edu Wed Oct 20 08:41:25 2004 From: Jacqueline.Miller <@t> UTSouthwestern.edu (Jacqueline Miller) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] processing schedule Message-ID: Aren't there laws that protect the mentally challenged from being used in research? >>> "Rice, Michael" 10/20/04 8:35 AM >>> Maybe we should use politicians for research instead of baboons -----Original Message----- From: Barry R Rittman [mailto:Barry.R.Rittman@uth.tmc.edu] Sent: Wednesday, October 20, 2004 9:06 AM To: histonet Subject: RE: [Histonet] processing schedule If so we would have a lot less politicians. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rice, Michael Sent: Wednesday, October 20, 2004 7:40 AM To: renton louise mrs; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] processing schedule arent baboons an endangered species? -----Original Message----- From: renton louise mrs [mailto:rentonlf@bru.wits.ac.za] Sent: Wednesday, October 20, 2004 7:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] processing schedule Hello, could someone please advise me as to how to adjust a processing schedule for a "dip & dunk" processor to one in which heat and vacuum is an option? I am processing non decalcified baboon bone - specifically full thickness jaw (no teeth, thank goodness) and skull? The thickness obviously varies from 0.5 - 1cm and the area is about 2cm square. In the didp & dunk we generally left the specimen for 24hrs per beaker, but I am sure these times could be reduced. Thank you in advance & best regards Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- _______________________________________________ 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 ---------------------- Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pmarcum <@t> polysciences.com Wed Oct 20 08:39:06 2004 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] processing schedule In-Reply-To: <3BC92F29BE821745AB15E04C98EE028D69361C@HCH2KMAIL.holy-cross.com> Message-ID: <000c01c4b6aa$2ba8e100$4f00a8c0@PMARCUM2K> That is a wonderful idea we just may not be able to do much brain research on them. Pam Marcum (Not necessarily the view of the company) > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rice, > Michael > Sent: Wednesday, October 20, 2004 9:36 AM > To: Barry R Rittman; histonet > Subject: RE: [Histonet] processing schedule > > > Maybe we should use politicians for research instead of baboons > > -----Original Message----- > From: Barry R Rittman [mailto:Barry.R.Rittman@uth.tmc.edu] > Sent: Wednesday, October 20, 2004 9:06 AM > To: histonet > Subject: RE: [Histonet] processing schedule > > > If so we would have a lot less politicians. > Barry > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rice, > Michael > Sent: Wednesday, October 20, 2004 7:40 AM > To: renton louise mrs; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] processing schedule > > arent baboons an endangered species? > > > > -----Original Message----- > > From: renton louise mrs [mailto:rentonlf@bru.wits.ac.za] > > Sent: Wednesday, October 20, 2004 7:41 AM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] processing schedule > > > > > > Hello, > > > > could someone please advise me as to how to adjust a processing schedule > for a "dip & dunk" processor to one in which heat and vacuum is an > option? > > > > I am processing non decalcified baboon bone - specifically full > thickness jaw (no teeth, thank goodness) and skull? > > The thickness obviously varies from 0.5 - 1cm and the area is about 2cm > square. In the didp & dunk we generally left the specimen for 24hrs per > beaker, but I am sure these times could be reduced. > > > > Thank you in advance & best regards > > Louise Renton > > Bone Research Unit > > University of the Witwatersrand > > Johannesburg > > South Africa > > .......so what IS the speed of dark? > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > ---------------------- > Confidentiality Notice: This e-mail message, including any attachments, > is for the sole use of the intended recipient(s) and may contain > confidential and privileged information. Any unauthorized review, use, > disclosure, or distribution is prohibited. If you are not the intended > recipient, please contact the sender by reply e-mail and destroy all > copies of the original. > ---------------------- > > > > _______________________________________________ > 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 > > > ---------------------- > Confidentiality Notice: This e-mail message, including any > attachments, is for the sole use of the intended recipient(s) and > may contain confidential and privileged information. Any > unauthorized review, use, disclosure, or distribution is > prohibited. If you are not the intended recipient, please contact > the sender by reply e-mail and destroy all copies of the original. > ---------------------- > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Michael.Rice <@t> holy-cross.com Wed Oct 20 08:41:51 2004 From: Michael.Rice <@t> holy-cross.com (Rice, Michael) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] processing schedule Message-ID: <3BC92F29BE821745AB15E04C98EE028D69361D@HCH2KMAIL.holy-cross.com> all rules are made to be broken -----Original Message----- From: Jacqueline Miller [mailto:Jacqueline.Miller@UTSouthwestern.edu] Sent: Wednesday, October 20, 2004 9:41 AM To: Rice, Michael; histonet@lists.utsouthwestern.edu; Barry.R.Rittman@uth.tmc.edu Subject: RE: [Histonet] processing schedule Aren't there laws that protect the mentally challenged from being used in research? >>> "Rice, Michael" 10/20/04 8:35 AM >>> Maybe we should use politicians for research instead of baboons -----Original Message----- From: Barry R Rittman [mailto:Barry.R.Rittman@uth.tmc.edu] Sent: Wednesday, October 20, 2004 9:06 AM To: histonet Subject: RE: [Histonet] processing schedule If so we would have a lot less politicians. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rice, Michael Sent: Wednesday, October 20, 2004 7:40 AM To: renton louise mrs; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] processing schedule arent baboons an endangered species? -----Original Message----- From: renton louise mrs [mailto:rentonlf@bru.wits.ac.za] Sent: Wednesday, October 20, 2004 7:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] processing schedule Hello, could someone please advise me as to how to adjust a processing schedule for a "dip & dunk" processor to one in which heat and vacuum is an option? I am processing non decalcified baboon bone - specifically full thickness jaw (no teeth, thank goodness) and skull? The thickness obviously varies from 0.5 - 1cm and the area is about 2cm square. In the didp & dunk we generally left the specimen for 24hrs per beaker, but I am sure these times could be reduced. Thank you in advance & best regards Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- _______________________________________________ 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 ---------------------- Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- From jcox90 <@t> yahoo.com Wed Oct 20 08:45:59 2004 From: jcox90 <@t> yahoo.com (Jill Cox) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Thanks!!! My Microtomy service is up and running Message-ID: <20041020134559.14116.qmail@web40901.mail.yahoo.com> Hello Netters, I want to thank you all again for all of the VERY useful tips on starting up my microtomy service. It’s up and running and I have my first client. The website is up also, www.precisionmicrotomy.com . I will be making a link for the email and a couple other changes. If anyone has any ideas or comments I would love to hear them. Jill Sorry for the first message , forgot the subject line. Jill Cox HT (ASCP) --------------------------------- Do you Yahoo!? Yahoo! Mail Address AutoComplete - You start. We finish. From mbryhan <@t> NORTHERNHEALTH.ORG Wed Oct 20 08:49:56 2004 From: mbryhan <@t> NORTHERNHEALTH.ORG (Mary Bryhan) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Lost specimens Message-ID: All- Our pathologists have given us the task of benchmarking "lost specimens". We are trying to find out what other labs do when something does not survive processing. Any info. would be greatly appreciated. Mary Bryhan From jcox90 <@t> yahoo.com Wed Oct 20 09:14:55 2004 From: jcox90 <@t> yahoo.com (Jill Cox) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Fwd: RE: typo in website Message-ID: <20041020141455.97749.qmail@web40907.mail.yahoo.com> Note: forwarded message attached. Jill Cox HT (ASCP) --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From lea <@t> lsr.auc.dk Wed Oct 20 09:18:06 2004 From: lea <@t> lsr.auc.dk (Lea Bjerre) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Silver staining protocol Message-ID: <000c01c4b6af$9e936b90$870ba8c0@trinepc> Hi. I?m searching for a general protein staining method (not including coomassie brillant blue). I thought maybe silver, because of very low amounts of the proteins. The dye is for a membrane coating, not a gel. Can you help me out? Regards Lea Bjerre From cbrayton <@t> bcm.tmc.edu Wed Oct 20 09:23:30 2004 From: cbrayton <@t> bcm.tmc.edu (BCM HUMAN RESOURCES) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Re: Histonet Digest, Vol 11, Issue 25 Message-ID: <200410201423.JAA27424@morpheus.corp.bcm.tmc.edu> This user is no longer with Baylor College of Medicine. Thanks, Baylor College of Medicine From funderwood <@t> mcohio.org Wed Oct 20 09:41:28 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:11 2005 Subject: [BULK] - RE: [Histonet] processing schedule Message-ID: Important information could be gained, however. Such as why there is the formation a rectolaryngeal fistula. Which leads to talking out of your a**. >>> "Pamela Marcum" 10/20/04 09:39AM >>> That is a wonderful idea we just may not be able to do much brain research on them. Pam Marcum (Not necessarily the view of the company) > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rice, > Michael > Sent: Wednesday, October 20, 2004 9:36 AM > To: Barry R Rittman; histonet > Subject: RE: [Histonet] processing schedule > > > Maybe we should use politicians for research instead of baboons > > -----Original Message----- > From: Barry R Rittman [mailto:Barry.R.Rittman@uth.tmc.edu] > Sent: Wednesday, October 20, 2004 9:06 AM > To: histonet > Subject: RE: [Histonet] processing schedule > > > If so we would have a lot less politicians. > Barry > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rice, > Michael > Sent: Wednesday, October 20, 2004 7:40 AM > To: renton louise mrs; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] processing schedule > > arent baboons an endangered species? > > > > -----Original Message----- > > From: renton louise mrs [mailto:rentonlf@bru.wits.ac.za] > > Sent: Wednesday, October 20, 2004 7:41 AM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] processing schedule > > > > > > Hello, > > > > could someone please advise me as to how to adjust a processing schedule > for a "dip & dunk" processor to one in which heat and vacuum is an > option? > > > > I am processing non decalcified baboon bone - specifically full > thickness jaw (no teeth, thank goodness) and skull? > > The thickness obviously varies from 0.5 - 1cm and the area is about 2cm > square. In the didp & dunk we generally left the specimen for 24hrs per > beaker, but I am sure these times could be reduced. > > > > Thank you in advance & best regards > > Louise Renton > > Bone Research Unit > > University of the Witwatersrand > > Johannesburg > > South Africa > > .......so what IS the speed of dark? > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > ---------------------- > Confidentiality Notice: This e-mail message, including any attachments, > is for the sole use of the intended recipient(s) and may contain > confidential and privileged information. Any unauthorized review, use, > disclosure, or distribution is prohibited. If you are not the intended > recipient, please contact the sender by reply e-mail and destroy all > copies of the original. > ---------------------- > > > > _______________________________________________ > 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 > > > ---------------------- > Confidentiality Notice: This e-mail message, including any > attachments, is for the sole use of the intended recipient(s) and > may contain confidential and privileged information. Any > unauthorized review, use, disclosure, or distribution is > prohibited. If you are not the intended recipient, please contact > the sender by reply e-mail and destroy all copies of the original. > ---------------------- > > > _______________________________________________ > 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 From mlgiebel <@t> mail2.vcu.edu Wed Oct 20 09:41:48 2004 From: mlgiebel <@t> mail2.vcu.edu (Mary Lee Giebel) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Lost specimens In-Reply-To: Message-ID: <200410201441.KAA20375@arrakis.vcu.edu> Cry? ------------------- > All- > > Our pathologists have given us the task of benchmarking "lost specimens". We are trying to find out what other labs do when something does not survive processing. Any info. would be greatly appreciated. > > Mary Bryhan > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From contact <@t> excaliburpathology.com Wed Oct 20 09:56:27 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Eosin, phloxine, & OG Message-ID: <20041020145627.24542.qmail@web50302.mail.yahoo.com> I have used a product called Treosin for years that contains eosin, phloxine, and orange G. After deparaffination and hematoxylin staining, rinse, differentiate in acid alcohol, rinse, blue in ammonia water, rinse, then go to 95% alcohol, then directly into Treosin for only 15 seconds, then dehydrate in 100% alcohol, clear in xylene, and coverslip. http://www.statlab.com/product.asp?catalog%5Fname=StatLab&category%5Fname=Stains&product%5Fid=SL93%2D1 Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From Terry.Marshall <@t> rothgen.nhs.uk Wed Oct 20 09:55:45 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Lost specimens Message-ID: That's one of the easiest questions ever posed here. I simply say so in the report. If it is possible to give some indication of the likely cause, that is mentioned. So a report may go something like.... "Regret material did not survive processing - possibly due to its being mucin only." Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Mary Bryhan [mailto:mbryhan@NORTHERNHEALTH.ORG] Sent: 20 October 2004 14:50 To: histonet@pathology.swmed.edu Cc: Ellen Clausen-Simon; Shawn Leslie; Lee Anne Whipp Subject: [Histonet] Lost specimens All- Our pathologists have given us the task of benchmarking "lost specimens". We are trying to find out what other labs do when something does not survive processing. Any info. would be greatly appreciated. Mary Bryhan _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RSRICHMOND <@t> aol.com Wed Oct 20 10:03:17 2004 From: RSRICHMOND <@t> aol.com (RSRICHMOND@aol.com) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Re: Eosin, phloxine, and OG? Message-ID: <67B75EB6.6930B49F.0BF1BB09@aol.com> A reader identified only as Deb asks: >>is anyone familiar with an eosin/phloxine that mixes into it some OG(for H+E staining? The conversation came up recently to "amp" the three shades of red in H+E staining. It was the first time I'd ever heard of adding Orange G into eosin/phloxine solution. Are there any labs doing this?<< Such mixtures were in occasional use in the 1960's, and at least one was commercially available from a company in Texas. I've had only minimal experience with them and didn't find them greatly helpful in surgical pathology, but I'm not sure I ever saw one done by a histotech who was willing to look at a slide under the microscope. Trichrome stains were actually used by some surgical pathologists a long time ago as their routine stain, and I think the very beautiful hematoxylin-phloxin-saffron stain may still be in use as a routine stain in a few labs. In the 1930's Chandler Foot at New York Hospital (Cornell Medical Center) used a green trichrome as a routine stain. A few of his slides survived in old teaching collections there in 1968. Bob Richmond Knoxville TN and Gastonia NC From Terry.Marshall <@t> rothgen.nhs.uk Wed Oct 20 10:08:59 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Eosin, phloxine and OG? Message-ID: Come on John - you can't get away with half a story:-) If H&E isn't a good routine stain, what is (and how many years to get used to it)? Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: John Kiernan [mailto:jkiernan@uwo.ca] Sent: 20 October 2004 06:26 To: WWmn916@aol.com Cc: histonet@pathology.swmed.edu Subject: Re: [Histonet] Eosin, phloxine and OG? If you look at your H&E slides between staining and clearing & coverslipping you will find that eosin Y (CI 45380) can impart more than 3 colours to RBC, smooth muscle cytoplasm, other cytoplasms and collagen. Adding phloxin or orange G to an eosin Y counterstain may extend the spectrum, but don't expect clear-cut results from slides stained by a machine. The H&E method needs human judgement and adjustments before applying a coverslip. For this reason and others H&E is not a good "routine" staining method. It's all there in the peer-reviewed literature and in most histotechnology textbooks less than 25 years old. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ > (for H+E staining)? > The conversation came up recently to "amp" the three shades of red in H+E > staining. It was the first time I'd ever heard of adding Orange G into > eosin/phloxing solution. Are there any labs doing this? > > Thanks:-) > Deb > _______________________________________________ > 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 From contact <@t> excaliburpathology.com Wed Oct 20 10:22:08 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Microtomy service Message-ID: <20041020152208.35658.qmail@web50302.mail.yahoo.com> Congrats Jill and good luck !!!! I too have started my own histology service lab, however I specialize in whole eye tissue and am doing work for researchers on mice, rats, and rabbits. I am CLIA certified to do human tissue also. SO if anyones lab is not setup to do whole eyes which require special fixation, clearing agents, and paraffin, I would be happy to help. I receive specimens via FedEx all the time. Just call first so I may discuss special fixation they need and how to ship. Paula Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From bhewlett <@t> cogeco.ca Wed Oct 20 10:23:45 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] RE: Controls for Immunos fixed in B-Plus References: <83AACDB0810528418AA106F9AE9B7F7E278518@sjhaexc02.sjha.org> Message-ID: <003b01c4b6b8$ca2e5bd0$6400a8c0@mainbox> Joyce, In short, YES! GLP and NCCLS IHC guidelines indicate that tissue controls should be fixed and processed in the same way as the patient tissue. You should make your own comparisons regarding IHC performance on B-Plus fixed material vs your routine fixative. Regards, Bryan ----- Original Message ----- From: "Weems, Joyce" To: "Weems, Joyce" ; "Histonet" Sent: Wednesday, October 20, 2004 9:05 AM Subject: [Histonet] RE: Controls for Immunos fixed in B-Plus > I haven't heard from anyone! Let me ask again. > > Thanks, > Joyce > > -----Original Message----- > From: Weems, Joyce > Sent: Friday, October 15, 2004 11:28 AM > To: 'Histonet' > Subject: Controls for Immunos fixed in B-Plus > > > Hi Everyone, > > Do you use B-Plus fixed control tissue for immunos done on B-Plus fixed patient tissue? If so, how does it compare to formalin or B-5? > Thanks in advance. j > And happy weekend to everyone! j > > Joyce Weems > Pathology Manager > Saint Joseph's Hospital > 5665 Peachtree Dunwoody Rd NE > Atlanta, GA 30342 > 404-851-7376 - Phone > 404-851-7831 - Fax > > > > > Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. > Thank you. Saint Josephs Health System, Inc. > > ---------------------------------------------------------------------------- ---- > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Hared <@t> FGHI.COM Wed Oct 20 10:30:26 2004 From: Hared <@t> FGHI.COM (Harris, Ed) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Microwave retrieval Message-ID: Can anyone give me the pro's and con's of steamer/pressure (Decloaker) vs. microwave retrieval ? Our Doc wants us to change from the Decloaker to microwave. Is a household microwave adequate or do we need a lab grade unit? Does the microwave save time and improve turn around time? Thanks in advance Ed Harris HT ASCP MT ABB This E-mail message (including attachments, if any) is intended for the use of the individual or entity to which it is addressed and may contain information that is privileged, proprietary, confidential and exempt from disclosure. If you are not the intended recipient, you are notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender and erase this E-mail message immediately. From wayneholland1959 <@t> msn.com Wed Oct 20 10:44:16 2004 From: wayneholland1959 <@t> msn.com (Wayne Holland) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Hydrophobic properties on red control box slides. Message-ID: Has anyone experienced using an automated immuno system that maintains the slides on a horizontal plain during the run, having possible hydrophobic properties. This prevents the antibody and other reagents from being dispersed evenly on the slide. The reagents on this system are dispensed in the area of the control box (with the control box being painted on the underside of the slide). However when these slide are shipped, they are boxed back to front laying on one another of course, this causing the paint properties to be transferred to the slide in front if it. I have found that I have had to run the coated slides through Xylene and alcohol, then air dry under a hood before we mount the tissue. Has anyone experienced this being a problem? Once we started this additional step, all the problems of the tissue in the control box staining and not the patient has been eliminated. PLEASE HELP. The people that make the glass think I'm crazy. _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From blaesse <@t> rhrk.uni-kl.de Wed Oct 20 10:57:39 2004 From: blaesse <@t> rhrk.uni-kl.de (Peter Blaesse) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] FLAG antibody Message-ID: Hi, has anyone experience with antibodies which detect the FLAG protein tag? I want to analyze the expression of a FLAG-tagged transgene in brainstem sections of mice. I have tried one FLAG-ab from Kodak/Sigma, but it didn?t work (after perfusion with 4% PFA). I have not tried antigen-retrieval, due to the fact, that I was quite unhappy with the tissue quality after the procedure in the past. There are other FLAG-abs (e.g. Chemicon MAB3118), but I am not interested in testing them all. Thanks for your help. peter From wayneholland1959 <@t> msn.com Wed Oct 20 11:00:29 2004 From: wayneholland1959 <@t> msn.com (Wayne Holland) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Hydrophobic properties on red control box slides. Message-ID: Let me make it clear that the manufacter is working closely with me to resolve this problem, they have offered much help in this matter. They are also examining the lots that were used. Maybe I thought I was crazy. Your input please! >From: "Wayne Holland" >To: Histonet@lists.utsouthwestern.edu >Subject: [Histonet] Hydrophobic properties on red control box slides. >Date: Wed, 20 Oct 2004 11:44:16 -0400 > >Has anyone experienced using an automated immuno system that maintains the >slides on a horizontal plain during the run, having possible hydrophobic >properties. This prevents the antibody and other reagents from being >dispersed evenly on the slide. The reagents on this system are dispensed in >the area of the control box (with the control box being painted on the >underside of the slide). However when these slide are shipped, they are >boxed back to front laying on one another of course, this causing the paint >properties to be transferred to the slide in front if it. I have found that >I have had to run the coated slides through Xylene and alcohol, then air >dry under a hood before we mount the tissue. Has anyone experienced this >being a problem? Once we started this additional step, all the problems of >the tissue in the control box staining and not the patient has been >eliminated. PLEASE HELP. The people that make the glass think I'm crazy. > >_________________________________________________________________ >Don’t just search. Find. Check out the new MSN Search! >http://search.msn.click-url.com/go/onm00200636ave/direct/01/ > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! hthttp://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From bill501 <@t> mindspring.com Wed Oct 20 11:03:22 2004 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Lost specimens In-Reply-To: References: Message-ID: At 3:55 PM +0100 10/20/04, Marshall Terry Dr, Consultant Histopathologist wrote: >"Regret material did not survive processing - possibly due to its >being mucin only." I agree. Honesty is the best policy. On some specimens, eg, minute <0.05 cm specks, I will put into the gross the "This specimen may not survive processing". I also call the Doc and inform them. If a cassette opens, we filter all processor solutions and have almost always found the specimen. I also add that this was done. I suspect that we all have had badly processed tissue, which just does not cut or stain well. The worst example I can think of is a breast biopsy which cut poorly and would not stain with H&E, PAS, trichrome or Giemsa. I could make some guesses - purulent material was present, as was some granulation tissue. But, I could not really r/o carcinoma. In this case I stated the problem, what we tried etc and called the Doc. I have wondered if the alcohols were mixed up, but we never did find the cause. The worst thing to do is to pretend nothing happened or criminally make up a diagnosis. Bill From tgoodpas <@t> seattlecca.org Wed Oct 20 12:33:36 2004 From: tgoodpas <@t> seattlecca.org (Goodpaster, Tracy A) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Dog antibody search Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B948B39DB0@wala01.seattlecca.org> Hello Jackie O' I use Cleaved Caspase-3 from Cell Signaling (Clone Asp175, cat.#:9661L) on dog lymph node and gut with great success. I use it at 1:100 with Dako Target Retrieval Solution pH 6.0. I use the same solution for the serum block as antibody diluent, which is a mixture of Dako washing buffer and 20% serum (10% goat serum, 5% human serum, 5% dog serum). The secondary is Vector's Biotinylated goat anti-rabbit BA-1000. The tertiary is Vector's R.T.U. Vectastain Kit (PK-7100). I tried Dako's Envision kit (great for so many antibodies) but it gave a slightly weaker stain that was not as crisp. The DAB+ and Hematoxalin were from Dako (K4011 and S3301 respectively). I hope this will help you. Tracy Goodpaster -----Original Message----- From: Jackie.O'Connor@abbott.com [mailto:Jackie.O'Connor@abbott.com] Sent: Tuesday, October 19, 2004 5:01 AM To: Histonet Subject: [Histonet] Dog antibody search Hi guys - Can anyone steer me towards a supplier for Caspase-3, MPM-2, and Ki-67 primary antibodies that will work for IHC on FFPE Dog?? Thanks much! Jackie O' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. From Marie-Helene.Lavergne <@t> mdsps.com Wed Oct 20 12:58:09 2004 From: Marie-Helene.Lavergne <@t> mdsps.com (Marie-Helene.Lavergne@mdsps.com) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Sectionnong whole-body rats with a BRIGHT 8250 Cryostat Message-ID: Hi everyone, I'm using a Bright 8250 cryostat to do rat whole-body sections for QWBA analysis. Since I started using the microtome, (even with blood standards blocks or a rat carcass) sometimes (1 time/10), the knife appears to slip on the block at the begginning of the block and start to section at the end. I tried two type of knife (D-profile and tungsten) and many angles (generaly 10 degree for the D-profile and 20 for tungsten) I tried to adjust the slide tensioners. I tried two types of CMC. I tried different temperature (-20 c, -25 c) and it still doesn't work. WHAT IS THE PROBLEM???? Please, HELP ME, I'm trying to solve this problem since many months. Marie-Helene Lavergne Analyst DMPK MDS Pharma [1]marie-helene.lavergne@mdsps.com References 1. file://localhost/tmp/mne@mdsps.com From carrie <@t> ethossearch.com Wed Oct 20 13:42:46 2004 From: carrie <@t> ethossearch.com (Carrie Roy) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Job Opportunity - Northern California Message-ID: This is an opportunity to join the worldwide leader in reagent, consumable, and equipment sales to the anatomical pathology market at a time when the company is growing and expanding its product offerings. They are seeking a histology professional with a strong technical background to cover a sales territory in the Northern California/Northern Nevada. This is a position where you can utilize your scientific expertise to assist others by representing products into the diagnostic lab environment. This company is a worldwide developer and producer of instruments and supplies for anatomical pathology and other key laboratory disciplines. The company develops and produces a complete portfolio of consumables and supplies for the histology, cytology and hematology lab community and they are the leading histology and cytology supplies manufacturer for the North American market. Base salary $45 +/-, with $25-30K possible in commission first year (current rep is making well over this!) Company car will be either an SUV or van Position requires light travel and is open due to promotion of the current rep into another product line. If you or anyone you know is interested in this, or any other scientific related position, please feel free to contact me at: Carrie Roy Ethos Search Consultants 4717 Vanderhill Road Torrance, CA 90505 carrie@ethossearch.com 310-791-4396 From info <@t> instrumedics.com Wed Oct 20 13:51:18 2004 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] infectious frozens Message-ID: <000f01c4b6d5$cba935d0$6401a8c0@INSTRUMEDICS22> The Cryo-Vac-Away vacuum system, which can be installed in your cryostat, is not a true decontamination system, but goes a long way in protecting you from infectious trimmings in the cryostat The trimmings created while facing off the block are suctioned away as they are generated at the blockface. The trimmings are captured in a cold primary filter inside the cryostat and pathogens are captured by secondary viral/bacterial filter downstream of the primary filter. The cryostat remains spotless! The details can be found on our web site www.instrumedics.com Bernice Instrumedics 800-237-2772 From bills <@t> icpmr.wsahs.nsw.gov.au Wed Oct 20 16:33:26 2004 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Hydrophobic properties on red control box slides. In-Reply-To: Message-ID: <001301c4b6ec$6f815320$83a7080a@wsahs.nsw.gov.au> Wayne, I am sure the Histonet archives will give you a large amount of information about this. We had problems as well and have been in correspondence with the Menzel Group in Germany about this. They are looking into the problem and agree there may be a problem with some batches (this appears to be worldwide). Is it only with slides which have had EDTA retrieval? We had slides on a parallel run, with the same pretreatment and antibody, which have stained with haematoxylin only, others with no staining with anything, others with partial staining across the section. The people from Menzel asked us to send them slides, both stained! and unused, from the batch (always only one batch) which gave these results, they agree that there is something peculiar about them. It happens infrequently this is the second time in two years and not all slides are affected. Ah the joy of being a Histotech Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Wayne Holland Sent: Thursday, 21 October 2004 2:00 AM To: wayneholland1959@msn.com; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Hydrophobic properties on red control box slides. Let me make it clear that the manufacter is working closely with me to resolve this problem, they have offered much help in this matter. They are also examining the lots that were used. Maybe I thought I was crazy. Your input please! >From: "Wayne Holland" >To: Histonet@lists.utsouthwestern.edu >Subject: [Histonet] Hydrophobic properties on red control box slides. >Date: Wed, 20 Oct 2004 11:44:16 -0400 > >Has anyone experienced using an automated immuno system that maintains the >slides on a horizontal plain during the run, having possible hydrophobic >properties. This prevents the antibody and other reagents from being >dispersed evenly on the slide. The reagents on this system are dispensed in >the area of the control box (with the control box being painted on the >underside of the slide). However when these slide are shipped, they are >boxed back to front laying on one another of course, this causing the paint >properties to be transferred to the slide in front if it. I have found that >I have had to run the coated slides through Xylene and alcohol, then air >dry under a hood before we mount the tissue. Has anyone experienced this >being a problem? Once we started this additional step, all the problems of >the tissue in the control box staining and not the patient has been >eliminated. PLEASE HELP. The people that make the glass think I'm crazy. > >_________________________________________________________________ >Don?t just search. Find. Check out the new MSN Search! >http://search.msn.click-url.com/go/onm00200636ave/direct/01/ > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! hthttp://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. From ccrowder <@t> mail.vetmed.lsu.edu Wed Oct 20 16:51:48 2004 From: ccrowder <@t> mail.vetmed.lsu.edu (Cheryl Crowder) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] PLP Fixation Message-ID: We have a research project in which all the tissues are fixed in periodate-lysine-paraformaldehyde (PLP). The question has arisen as to how long in advance to use can the solution be made? and the solution turns yellow after approx. 24 hours. Is this solution still usable? Thank you for your help. Cheryl Cheryl Crowder, BA, HTL(ASCP) Chief Technologist Anatomic Pathology Department of Pathobiological Sciences School of Veterinary Medicine Louisiana State University Skip Bertman Drive Baton Rouge, LA 70803 225-578-9734 FAX: 225-578-9720 From katri <@t> cogeco.ca Wed Oct 20 17:11:24 2004 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Microwave retrieval References: Message-ID: <009001c4b6f1$bd0743c0$6a9a9618@Katri> Hi Ed, My experience: Microwave with coplin jars overboils and you need to add buffer periodically (drying of tissue can happen), hot spots in microwave often result in uneven staining. We used microwavable pressurecooker (Dako) for many years and results improved, but we still had uneven staining occasionally. Now we are using the Decloaker and results are more reproducible and a log of temperature/pressure can be kept. However, we did have to do some adjustments in primary antibody dilutions. Concentrations had to be increased with some, therefore costing more. Steamer, I personally have not used, but my understanding is that reproducibility is pretty good, but the procedure takes a bit more time. My ten cent's worth... Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Harris, Ed" To: Sent: Wednesday, October 20, 2004 11:30 AM Subject: [Histonet] Microwave retrieval Can anyone give me the pro's and con's of steamer/pressure (Decloaker) vs. microwave retrieval ? Our Doc wants us to change from the Decloaker to microwave. Is a household microwave adequate or do we need a lab grade unit? Does the microwave save time and improve turn around time? Thanks in advance Ed Harris HT ASCP MT ABB This E-mail message (including attachments, if any) is intended for the use of the individual or entity to which it is addressed and may contain information that is privileged, proprietary, confidential and exempt from disclosure. If you are not the intended recipient, you are notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender and erase this E-mail message immediately. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Wed Oct 20 18:12:43 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Lost specimens Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E0D9@simba.kids> Some specimens, eg mucoid aspirates or waxy ear accumulations, do not survive processing. In these cases the processing cassette containing the paper or foam, which was supposed to contain the specimen, is allowed to cool and the cassette is filed with the other cassettes belonging to the case (audit trail) and a comment such as "tissue did not survive processing" is included in the report. Regards, Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Mary Bryhan [mailto:mbryhan@NORTHERNHEALTH.ORG] Sent: Wednesday, 20 October 2004 11:50 PM To: histonet@pathology.swmed.edu Cc: Ellen Clausen-Simon; Shawn Leslie; Lee Anne Whipp Subject: [Histonet] Lost specimens All- Our pathologists have given us the task of benchmarking "lost specimens". We are trying to find out what other labs do when something does not survive processing. Any info. would be greatly appreciated. Mary Bryhan _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From amosbrooks <@t> earthlink.net Wed Oct 20 19:00:20 2004 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Microwave retrieval In-Reply-To: <200410201000.1ckjPx77V3NZFmQ0@tanager.mail.pas.earthlink.n et> References: <200410201000.1ckjPx77V3NZFmQ0@tanager.mail.pas.earthlink.net> Message-ID: <6.0.0.22.0.20041020194855.01c78360@pop.earthlink.net> Ed, I strongly recommend that you don't try to put a coplin jar of retrieval solution with slides in it in the microwave and expect any consistant results. What happens with this method is you get a spike of heat then a temperature drop. Very often there is boil over and your slides dry out. Ideally you should be looking for an even temperature for a defined length of time that can be well monitored. For this reason if you *must* use a microwave it should be a lab grade oven that has consistant wattage and temperature control. It would also be good to use one of the microwave pressure cookers to improve the consistency. (I think Biocare Medical has them ... the guys that sell Borg Decloaker). My ultimate recommendation is to stick to the tried and true waterbath or steamer method. It may take a bit longer but the consistancy is better and there is far less variables in the procedure. Just my opinion, You know what they say 'bout opinions, Amos Brooks At 01:00 PM 10/20/2004, you wrote: >Message: 10 >Date: Wed, 20 Oct 2004 11:30:26 -0400 >From: "Harris, Ed" >Subject: [Histonet] Microwave retrieval >To: "'histonet@lists.utsouthwestern.edu'" > >Message-ID: >Content-Type: text/plain; charset="iso-8859-1" > >Can anyone give me the pro's and con's of steamer/pressure (Decloaker) vs. >microwave retrieval ? >Our Doc wants us to change from the Decloaker to microwave. Is a household >microwave adequate or do we need a lab grade unit? >Does the microwave save time and improve turn around time? > >Thanks in advance >Ed Harris HT ASCP MT ABB From ctsblack <@t> capeheart.uct.ac.za Thu Oct 21 01:28:10 2004 From: ctsblack <@t> capeheart.uct.ac.za (Melanie Black) Date: Fri Sep 16 15:24:11 2005 Subject: Fwd: [Histonet] RE: AP problem Message-ID: >I have also had rod-like crystals in my Alk phos/NBT system. I have >shown them to the company rep, who has no idea what they might be. >With so much written about it here...I think DAKO should sit up and >take a look. Melanie Black > >Thanks all for the cat scratch advice. I am trying it with HRP- AEC. >Will report result. >Jo > > >>>> "C.M. van der Loos" 10/17/04 02:20PM >>> >Dear Joanne, > >Usually, AP staining doesn't show up in a crystal-like precipitate, >but I have seen it a couple of times. Never found out what was the >cause of it. Be sure that after your IHC procedure and before the AP >visualization, the sections are washed at with a Tris-HCl pH8.5 >buffer. Phosphate ions from PBS will inhibit your AP reaction. You >also may mix the Permanent Red components first and than press >immediately through a syringe-filter to remove all bigger particles >in the AP solution. >Have you viewed your slides under a fluorescence microscope? A small >amount of Permanent Red reaction product also gives a bright red >fluorescence signal when using the rhodamine filter pack (green >light). >Good suggestion by Richard to use a tonsil as a negative control. >But, how about running the good old control blanc slide without any >primary antibody? >Hope this helps. > >Chris van der Loos, PhD >Dept. of Pathology >Academical Medical Center >Amsterdam - The Netherlands > >----- Original Message ----- >>From "Joanne Mauger" >Date Fri, 15 Oct 2004 15:45:05 -0400 >To histonet@lists.utsouthwestern.edu, jkiernan@uwo.ca >Subject Re: [Histonet] IHC withquantum dots >Hi group, >Can anyone help me with suggestions-I am working up the monoclonal >Ab (H2A10) to cat scratch disease. Ab from Novus Biologicus. I have >a purchased control from Newcomer Supply. I am using permanent red >chromagen from Dako., with alk phos link also from Dako. I am >getting a nice specific staining of what looks like specs vs tiny >rods or spirochetes, but the control tissue does , by Warthin >Starry, look much more spirochete-ish. Can I trust this result? I >have also stained several suspected + cases and see the same dust >like + staining,but I am afraid it is teeny bits of precipitate. >Does this ring a bell? >Please help. >Thanks, >Jo > >----- Original Message ----- >>From "Richard Cartun" >Date Fri, 15 Oct 2004 16:45:57 -0400 >To ,, > >Subject Re: [Histonet] IHC withquantum dots >Have you tried staining a section of tonsil to see if you get the >same pattern of staining. I have found "precipitation" to be a >problem with alkaline phosphatase (PA) detection, although I am by >no means an expert when it comes to AP IHC. I use HRP/AEC for most >of my infectious disease studies including Bartonella henselae. >Don't forget that you may not see the normal morphology of the >bacterium with IHC. You are labeling epitopes on the surface of the >organism; you are not staining the entire cell wall. I have found >B. henselae to be one of the most difficult organisms to label (and >interpret) with IHC. Good luck! > >Richard > > >Richard W. Cartun, Ph.D. >Director, Immunopathology & Histology >Assistant Director, Anatomic Pathology >Hartford Hospital >80 Seymour Street >Hartford, CT 06102 > > > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Cardiovascular Research Unit Div. of Cardiothoracic Surgery Chris Barnard Building University of Cape Town Anzio Road Observatory 7925 Republic of South Africa Tel +27 21 406-6589 Cel +27 82 469-3352 Fax +27 21 448-5935 From rentonlf <@t> bru.wits.ac.za Thu Oct 21 01:52:02 2004 From: rentonlf <@t> bru.wits.ac.za (renton louise mrs) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] processing schedule / baboons Message-ID: <1098341522.970fb540rentonlf@bru.wits.ac.za> OOps, Seems like I opened a can of worms here. Papio Hamadryas, the Chacma or hamadryas baboon IS CITES listed. These are primarily a northern african and indian species. I should have said in my post that we use the non-human primate Papio Ursinus, which is NOT endangered and is a considerable pest to the farming community (similar to politicians). So please, smoothe all those ruffled sensibilities and try to give me some assistance with my request. Thank you -----Original Message----- From: "Fred Underwood" To: Date: Wed, 20 Oct 2004 10:41:28 -0400 Subject: [BULK] - RE: [Histonet] processing schedule Important information could be gained, however. Such as why there is the formation a rectolaryngeal fistula. Which leads to talking out of your a**. >>> "Pamela Marcum" 10/20/04 09:39AM >>> That is a wonderful idea we just may not be able to do much brain research on them. Pam Marcum (Not necessarily the view of the company) > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rice, > Michael > Sent: Wednesday, October 20, 2004 9:36 AM > To: Barry R Rittman; histonet > Subject: RE: [Histonet] processing schedule > > > Maybe we should use politicians for research instead of baboons > > -----Original Message----- > From: Barry R Rittman [mailto:Barry.R.Rittman@uth.tmc.edu] > Sent: Wednesday, October 20, 2004 9:06 AM > To: histonet > Subject: RE: [Histonet] processing schedule > > > If so we would have a lot less politicians. > Barry > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rice, > Michael > Sent: Wednesday, October 20, 2004 7:40 AM > To: renton louise mrs; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] processing schedule > > arent baboons an endangered species? > > > > -----Original Message----- > > From: renton louise mrs [mailto:rentonlf@bru.wits.ac.za] > > Sent: Wednesday, October 20, 2004 7:41 AM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] processing schedule > > > > > > Hello, > > > > could someone please advise me as to how to adjust a processing schedule > for a "dip & dunk" processor to one in which heat and vacuum is an > option? > > > > I am processing non decalcified baboon bone - specifically full > thickness jaw (no teeth, thank goodness) and skull? > > The thickness obviously varies from 0.5 - 1cm and the area is about 2cm > square. In the didp & dunk we generally left the specimen for 24hrs per > beaker, but I am sure these times could be reduced. > > > > Thank you in advance & best regards > > Louise Renton > > Bone Research Unit > > University of the Witwatersrand > > Johannesburg > > South Africa > > .......so what IS the speed of dark? > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > ---------------------- > Confidentiality Notice: This e-mail message, including any attachments, > is for the sole use of the intended recipient(s) and may contain > confidential and privileged information. Any unauthorized review, use, > disclosure, or distribution is prohibited. If you are not the intended > recipient, please contact the sender by reply e-mail and destroy all > copies of the original. > ---------------------- > > > > _______________________________________________ > 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 > > > ---------------------- > Confidentiality Notice: This e-mail message, including any > attachments, is for the sole use of the intended recipient(s) and > may contain confidential and privileged information. Any > unauthorized review, use, disclosure, or distribution is > prohibited. If you are not the intended recipient, please contact > the sender by reply e-mail and destroy all copies of the original. > ---------------------- > > > _______________________________________________ > 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 Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? From lpwenk <@t> sbcglobal.net Thu Oct 21 03:56:20 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] FETAL BRAIN FIXATION Message-ID: <009101c4b74b$d63452c0$372ed445@domainnotset.invalid> Anyone have a good fixative for the very soft fetal brains? Our neuropathologist does not want to use Bouin, as the tissue turns yellow and photographs do not look the way they are supposed to. Another one of our pathologists remembers using an alcohol-acetone-formalin mixture years ago as a resident in another institution, but doesn't know the percentage of any of the reagents. I've tried several search engines and PubMed, and can't find anything. Thanks. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 From psanquin <@t> lugo.usc.es Thu Oct 21 04:12:40 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] FETAL BRAIN FIXATION In-Reply-To: <009101c4b74b$d63452c0$372ed445@domainnotset.invalid> Message-ID: <3.0.6.32.20041021111240.007de320@pop.lugo.usc.es> You can try with acetic-alcohol: Glacial acetic acid in absolute ethanol (5% v/v) Good luck, Pablo Sanchez At 04:56 a.m. 21/10/04 -0400, you wrote: >Anyone have a good fixative for the very soft fetal brains? > >Our neuropathologist does not want to use Bouin, as the tissue turns yellow >and photographs do not look the way they are supposed to. > >Another one of our pathologists remembers using an alcohol-acetone-formalin >mixture years ago as a resident in another institution, but doesn't know the >percentage of any of the reagents. > >I've tried several search engines and PubMed, and can't find anything. > >Thanks. > >Peggy A. Wenk, HTL(ASCP)SLS >William Beaumont Hospital >Royal Oak, MI 48073 > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Marjorie.Lehman <@t> unilever.com Thu Oct 21 06:18:47 2004 From: Marjorie.Lehman <@t> unilever.com (marjorie lehman) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Microwave retrieval Message-ID: Here is a Second Opinion! I tried using a (Household) microwave for a long time and was not happy. After I got my Decloaking Chamber from Biocare I never looked back! Marge -----Original Message----- From: Amos Brooks [SMTP:amosbrooks@earthlink.net] Sent: Wednesday, October 20, 2004 8:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave retrieval Ed, I strongly recommend that you don't try to put a coplin jar of retrieval solution with slides in it in the microwave and expect any consistant results. What happens with this method is you get a spike of heat then a temperature drop. Very often there is boil over and your slides dry out. Ideally you should be looking for an even temperature for a defined length of time that can be well monitored. For this reason if you *must* use a microwave it should be a lab grade oven that has consistant wattage and temperature control. It would also be good to use one of the microwave pressure cookers to improve the consistency. (I think Biocare Medical has them ... the guys that sell Borg Decloaker). My ultimate recommendation is to stick to the tried and true waterbath or steamer method. It may take a bit longer but the consistancy is better and there is far less variables in the procedure. Just my opinion, You know what they say 'bout opinions, Amos Brooks At 01:00 PM 10/20/2004, you wrote: >Message: 10 >Date: Wed, 20 Oct 2004 11:30:26 -0400 >From: "Harris, Ed" >Subject: [Histonet] Microwave retrieval >To: "'histonet@lists.utsouthwestern.edu'" > >Message-ID: >Content-Type: text/plain; charset="iso-8859-1" > >Can anyone give me the pro's and con's of steamer/pressure (Decloaker) vs. >microwave retrieval ? >Our Doc wants us to change from the Decloaker to microwave. Is a household >microwave adequate or do we need a lab grade unit? >Does the microwave save time and improve turn around time? > >Thanks in advance >Ed Harris HT ASCP MT ABB _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Thu Oct 21 07:15:08 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Microwave retrieval Message-ID: <83AACDB0810528418AA106F9AE9B7F7E278524@sjhaexc02.sjha.org> We were fortunate enough to have some left over capital dollars and have purchased the new Biogenex Retreival Microwave. It is wonderful - saves lots of time and gives consistency in staining! The only thing we would change is to be have smaller containers for instances when we have just a few slides to retreive. But we think its a winner for those of you are looking to purchase it. Cheers, Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of marjorie lehman Sent: Thursday, October 21, 2004 7:19 AM To: Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Microwave retrieval Here is a Second Opinion! I tried using a (Household) microwave for a long time and was not happy. After I got my Decloaking Chamber from Biocare I never looked back! Marge -----Original Message----- From: Amos Brooks [SMTP:amosbrooks@earthlink.net] Sent: Wednesday, October 20, 2004 8:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave retrieval Ed, I strongly recommend that you don't try to put a coplin jar of retrieval solution with slides in it in the microwave and expect any consistant results. What happens with this method is you get a spike of heat then a temperature drop. Very often there is boil over and your slides dry out. Ideally you should be looking for an even temperature for a defined length of time that can be well monitored. For this reason if you *must* use a microwave it should be a lab grade oven that has consistant wattage and temperature control. It would also be good to use one of the microwave pressure cookers to improve the consistency. (I think Biocare Medical has them ... the guys that sell Borg Decloaker). My ultimate recommendation is to stick to the tried and true waterbath or steamer method. It may take a bit longer but the consistancy is better and there is far less variables in the procedure. Just my opinion, You know what they say 'bout opinions, Amos Brooks At 01:00 PM 10/20/2004, you wrote: >Message: 10 >Date: Wed, 20 Oct 2004 11:30:26 -0400 >From: "Harris, Ed" >Subject: [Histonet] Microwave retrieval >To: "'histonet@lists.utsouthwestern.edu'" > >Message-ID: >Content-Type: text/plain; charset="iso-8859-1" > >Can anyone give me the pro's and con's of steamer/pressure (Decloaker) vs. >microwave retrieval ? >Our Doc wants us to change from the Decloaker to microwave. Is a household >microwave adequate or do we need a lab grade unit? >Does the microwave save time and improve turn around time? > >Thanks in advance >Ed Harris HT ASCP MT ABB _______________________________________________ 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 Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From vbaker60 <@t> yahoo.com Thu Oct 21 07:47:46 2004 From: vbaker60 <@t> yahoo.com (Victoria Baker) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Re: Lost specimens Message-ID: <20041021124747.10040.qmail@web50102.mail.yahoo.com> Mary, At time of submission, a notation would be made by the pathologist/PA -'MNSP' may not survive processing. As the majority of these specimens were aspirates or mucoid substances they would be coming out in our early basket and be on our biopsy embedding log with that notation. The embedder would wash the sponge or paper with wet paraffin, save the sponge and cassette top and make a block. We would still cut the block and submit to the pathologist. At time of sign out we would be told that nothing had survived. We would still keep the block and the slide would have DNSP written in red and filed. Because everything is entered on the computer the notation would automatically follow the patient/accession number for any future inquiries. We did not keep a separate log for these types of specimens as it never came up as an issue. Also, we kept all of our embedding waste for 1 week, so if there was any discrepancy we still had that to go back to. Vikki Baker __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From j_gorenstein <@t> yahoo.com Thu Oct 21 07:48:32 2004 From: j_gorenstein <@t> yahoo.com (Juile Gorenstein) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Anti-Dig options question for ISH Message-ID: <20041021124832.24478.qmail@web42002.mail.yahoo.com> Hello everyone, I was wondering, other than Roche, who else manufactures Anti-DIG antibodies. Does anyone have a favorite? (I am planning to use on Ventana Discovery) Thanks in advance! Julie Novartis Cambridge, MA --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From bhewlett <@t> cogeco.ca Thu Oct 21 08:32:32 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] FETAL BRAIN FIXATION References: <009101c4b74b$d63452c0$372ed445@domainnotset.invalid> Message-ID: <003a01c4b772$6bbcb320$6400a8c0@mainbox> Peggy, We faced the same issue a number of years ago. Our solution was to use a modified Brasil fixative. The pediatric pathologist was delighted with the results! The formula follows; Modified Brasil Fluid. Ethanol............................................. 700 mL 40% w/v Formaldehyde.....................100 mL Glacial Acetic acid..............................50 mL add D.water to make up to.................1000 mL Fetal brains are left to fix for 1-3 weeks, rinsed in 70% ETOH, then sliced and photographed. Selected blocks are processed using the same reagents and times as for NBF fixed material. Hope this helps, Bryan ----- Original Message ----- From: To: "Histonet" Sent: Thursday, October 21, 2004 4:56 AM Subject: [Histonet] FETAL BRAIN FIXATION > Anyone have a good fixative for the very soft fetal brains? > > Our neuropathologist does not want to use Bouin, as the tissue turns yellow > and photographs do not look the way they are supposed to. > > Another one of our pathologists remembers using an alcohol-acetone-formalin > mixture years ago as a resident in another institution, but doesn't know the > percentage of any of the reagents. > > I've tried several search engines and PubMed, and can't find anything. > > Thanks. > > Peggy A. Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From carmen_loiselle <@t> hotmail.com Thu Oct 21 08:44:28 2004 From: carmen_loiselle <@t> hotmail.com (carmen loiselle) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] NK1-C3 Message-ID: Good morning all, Is anybody out there familiar with the antibody " NK1-C3" ?? It's a putative melanoma antigen. If so, can you tell me where to find it, which company etc... Thanks in advance From jmitchell <@t> neurology.wisc.edu Thu Oct 21 09:03:04 2004 From: jmitchell <@t> neurology.wisc.edu (Mitchell (Jean)) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] PLP Fixation Message-ID: I use periodate-lysine-paraformaldehyde fixative for skin samples. I start with a lysine stock solution & a paraformaldehyde stock solution (each stable for 3 weeks). I make up the working solution with those stock solutions the day of use adding the periodate powder at that time. I have not experienced the solution turning yellow after 24 hours. If you would like more specifics on the solutions I use, I would gladly forward that information to you. Jean Mitchell, BS, HT (ASCP) University of Wisconsin Hospital & Clinics Department of Neurology Madison, WI -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Cheryl Crowder Sent: Wednesday, October 20, 2004 4:52 PM To: Histonet Subject: [Histonet] PLP Fixation We have a research project in which all the tissues are fixed in periodate-lysine-paraformaldehyde (PLP). The question has arisen as to how long in advance to use can the solution be made? and the solution turns yellow after approx. 24 hours. Is this solution still usable? Thank you for your help. Cheryl Cheryl Crowder, BA, HTL(ASCP) Chief Technologist Anatomic Pathology Department of Pathobiological Sciences School of Veterinary Medicine Louisiana State University Skip Bertman Drive Baton Rouge, LA 70803 225-578-9734 FAX: 225-578-9720 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From corporate_21 <@t> hotmail.com Thu Oct 21 09:07:12 2004 From: corporate_21 <@t> hotmail.com (corporate headquarters) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Thanks Message-ID: Just a thanks to Mad Dawg for all the info provided. Very interesting. _________________________________________________________________ On the road to retirement? Check out MSN Life Events for advice on how to get there! http://lifeevents.msn.com/category.aspx?cid=Retirement From convmcm <@t> cc.usu.edu Thu Oct 21 09:21:05 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Microwave retrieval new question In-Reply-To: <83AACDB0810528418AA106F9AE9B7F7E278524@sjhaexc02.sjha.org> Message-ID: <000d01c4b779$34090a30$4a737b81@Cygnus> I find this all interesting as my pathologist has suggested that I try the MW for our AR procedures. We were thinking about using it for proteinase K rather than using a citrate buffer or Tris. I would assume that the temp would be very important to control with the enzyme. So... For those of you who do MW AR, would you mind sharing your procedure? Thnks bunches!! *G* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 email: convmcm@cc.usu.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: Thursday, October 21, 2004 5:15 AM To: marjorie lehman; Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Microwave retrieval We were fortunate enough to have some left over capital dollars and have purchased the new Biogenex Retreival Microwave. It is wonderful - saves lots of time and gives consistency in staining! The only thing we would change is to be have smaller containers for instances when we have just a few slides to retreive. But we think its a winner for those of you are looking to purchase it. Cheers, Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of marjorie lehman Sent: Thursday, October 21, 2004 7:19 AM To: Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Microwave retrieval Here is a Second Opinion! I tried using a (Household) microwave for a long time and was not happy. After I got my Decloaking Chamber from Biocare I never looked back! Marge -----Original Message----- From: Amos Brooks [SMTP:amosbrooks@earthlink.net] Sent: Wednesday, October 20, 2004 8:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave retrieval Ed, I strongly recommend that you don't try to put a coplin jar of retrieval solution with slides in it in the microwave and expect any consistant results. What happens with this method is you get a spike of heat then a temperature drop. Very often there is boil over and your slides dry out. Ideally you should be looking for an even temperature for a defined length of time that can be well monitored. For this reason if you *must* use a microwave it should be a lab grade oven that has consistant wattage and temperature control. It would also be good to use one of the microwave pressure cookers to improve the consistency. (I think Biocare Medical has them ... the guys that sell Borg Decloaker). My ultimate recommendation is to stick to the tried and true waterbath or steamer method. It may take a bit longer but the consistancy is better and there is far less variables in the procedure. Just my opinion, You know what they say 'bout opinions, Amos Brooks At 01:00 PM 10/20/2004, you wrote: >Message: 10 >Date: Wed, 20 Oct 2004 11:30:26 -0400 >From: "Harris, Ed" >Subject: [Histonet] Microwave retrieval >To: "'histonet@lists.utsouthwestern.edu'" > >Message-ID: >Content-Type: text/plain; charset="iso-8859-1" > >Can anyone give me the pro's and con's of steamer/pressure (Decloaker) vs. >microwave retrieval ? >Our Doc wants us to change from the Decloaker to microwave. Is a household >microwave adequate or do we need a lab grade unit? >Does the microwave save time and improve turn around time? > >Thanks in advance >Ed Harris HT ASCP MT ABB _______________________________________________ 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 Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From Evelyn.Flynn <@t> childrens.harvard.edu Thu Oct 21 09:37:31 2004 From: Evelyn.Flynn <@t> childrens.harvard.edu (Flynn, Evelyn) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Microwave retrieval new question Message-ID: Dear Connie and others, In microwave antigen retrieval, I do the heating in three steps: 1. I heat a "water bath" filled with water for 5-10 minutes. I use a Rubbermaid plastic bowl, a 6 cup- size. 2. In PLASTIC Coplin jars sitting in the water bath I heat the retrieval solution for 5-10 minutes. In order for the jars to sit in the water bath, I fill the slots with blank slides (otherwise the jar would float). 3. Carefully with forceps I remove the blank slides and put in the slides with the sections. The solution should come to the top of the jar to allow for evaporation. Cover loosely to allow steam to escape. I microwave the jars in the waterbath for 15-20 min. Remove the jars from the waterbath, remove the covers, and allow to cool for 20-30 min. Good luck, Evelyn Flynn -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Connie McManus Sent: Thu 10/21/2004 10:21 AM To: 'Weems, Joyce'; 'marjorie lehman'; 'Amos Brooks'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Microwave retrieval new question I find this all interesting as my pathologist has suggested that I try the MW for our AR procedures. We were thinking about using it for proteinase K rather than using a citrate buffer or Tris. I would assume that the temp would be very important to control with the enzyme. So... For those of you who do MW AR, would you mind sharing your procedure? Thnks bunches!! *G* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 email: convmcm@cc.usu.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: Thursday, October 21, 2004 5:15 AM To: marjorie lehman; Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Microwave retrieval We were fortunate enough to have some left over capital dollars and have purchased the new Biogenex Retreival Microwave. It is wonderful - saves lots of time and gives consistency in staining! The only thing we would change is to be have smaller containers for instances when we have just a few slides to retreive. But we think its a winner for those of you are looking to purchase it. Cheers, Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of marjorie lehman Sent: Thursday, October 21, 2004 7:19 AM To: Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Microwave retrieval Here is a Second Opinion! I tried using a (Household) microwave for a long time and was not happy. After I got my Decloaking Chamber from Biocare I never looked back! Marge -----Original Message----- From: Amos Brooks [SMTP:amosbrooks@earthlink.net] Sent: Wednesday, October 20, 2004 8:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave retrieval Ed, I strongly recommend that you don't try to put a coplin jar of retrieval solution with slides in it in the microwave and expect any consistant results. What happens with this method is you get a spike of heat then a temperature drop. Very often there is boil over and your slides dry out. Ideally you should be looking for an even temperature for a defined length of time that can be well monitored. For this reason if you *must* use a microwave it should be a lab grade oven that has consistant wattage and temperature control. It would also be good to use one of the microwave pressure cookers to improve the consistency. (I think Biocare Medical has them ... the guys that sell Borg Decloaker). My ultimate recommendation is to stick to the tried and true waterbath or steamer method. It may take a bit longer but the consistancy is better and there is far less variables in the procedure. Just my opinion, You know what they say 'bout opinions, Amos Brooks At 01:00 PM 10/20/2004, you wrote: >Message: 10 >Date: Wed, 20 Oct 2004 11:30:26 -0400 >From: "Harris, Ed" >Subject: [Histonet] Microwave retrieval >To: "'histonet@lists.utsouthwestern.edu'" > >Message-ID: >Content-Type: text/plain; charset="iso-8859-1" > >Can anyone give me the pro's and con's of steamer/pressure (Decloaker) vs. >microwave retrieval ? >Our Doc wants us to change from the Decloaker to microwave. Is a household >microwave adequate or do we need a lab grade unit? >Does the microwave save time and improve turn around time? > >Thanks in advance >Ed Harris HT ASCP MT ABB _______________________________________________ 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 Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From abright <@t> brightinstruments.com Thu Oct 21 09:38:55 2004 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Sectionnong whole-body rats with a BRIGHT 8250 Cryostat Message-ID: Dear Marie-Helene, I do hope you have not been suffering these problems for to long, as the solution is simple. The knife angles are you are trying to section with are much too shallow and the specimen is being compressed by the rear end of the facet. The 'D' profile knife should be increased by 10? to 20? and the Tungsten Carbide Tipped knife increased by 6? to 26?. With these major changes to the knife angles you will find that the sharp edge of the knife will then be able to section, the correct sectioning temperature is -18 to -20?C. Best Regards Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: Marie-Helene.Lavergne@mdsps.com [mailto:Marie-Helene.Lavergne@mdsps.com] Sent: 20 October 2004 18:58 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Sectionnong whole-body rats with a BRIGHT 8250 Cryostat Hi everyone, I'm using a Bright 8250 cryostat to do rat whole-body sections for QWBA analysis. Since I started using the microtome, (even with blood standards blocks or a rat carcass) sometimes (1 time/10), the knife appears to slip on the block at the begginning of the block and start to section at the end. I tried two type of knife (D-profile and tungsten) and many angles (generaly 10 degree for the D-profile and 20 for tungsten) I tried to adjust the slide tensioners. I tried two types of CMC. I tried different temperature (-20 c, -25 c) and it still doesn't work. WHAT IS THE PROBLEM???? Please, HELP ME, I'm trying to solve this problem since many months. Marie-Helene Lavergne Analyst DMPK MDS Pharma [1]marie-helene.lavergne@mdsps.com References 1. file://localhost/tmp/mne@mdsps.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kerry.l.crabb <@t> gsk.com Thu Oct 21 09:44:56 2004 From: kerry.l.crabb <@t> gsk.com (kerry.l.crabb@gsk.com) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Kerry L Crabb/PharmRD/GSK is out of the office. Message-ID: I will be out of the office starting 21-Oct-2004 and will not return until 25-Oct-2004. Contact Eve about necropsy issue and contact Teresa about histology issues. I will respond to your message when I return. From contact <@t> excaliburpathology.com Thu Oct 21 10:15:47 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Microwave retrieval Message-ID: <20041021151547.74405.qmail@web50310.mail.yahoo.com> I have used a Black & Decker Rice Steamer for years for antigen retrieval and have had no problems. They are $30 at WalMart. Have one at home too, you can put a whole head of cauliflower in it. yum! Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From contact <@t> excaliburpathology.com Thu Oct 21 10:22:27 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Fetal brain Message-ID: <20041021152227.76947.qmail@web50310.mail.yahoo.com> When using Bouin's fixative, you are suppose to rinse several times in 70% alcohol, a couple hours each if whole specimen, to remove the yellow color before processing. Otherwise, the picric acid in Bouin's continues to "eat" your tissue and will interfere with staining. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From dellav <@t> musc.edu Thu Oct 21 10:28:43 2004 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] I may have a problem Message-ID: Greetings Linda or Herb, I have not been receiving histonet messages for some time. I believe this stems back to a time recently when the server was having difficulties. in any event. I thought I was un-subscribed and re-subscribed. I received a message stating that I am already subscribed. I'm uncertain what needs to be done next but I would be grateful for your advice or assistance. Sincerely Vinnie Della Speranza From shive003 <@t> umn.edu Thu Oct 21 10:38:44 2004 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] re: microwave retrieval Message-ID: <008701c4b784$0c702a20$78065486@vdl220FAC> I previously used plastic coplin jars set into a tupperware-type square dish, but also had occasional evaporating/boiling down of buffer and subsequent uncovering/drying out of tissues (even though I'd check and add to the buffer level after every 5 minutes). Now I use Tissue-Tek plastic containers, the rectangular-shaped ones that are used in the deparaffinization/rehydration Tissue-Tek system. They hold more buffer volume, and it seems that less buffer is evaporated/boiled off. I still check buffer levels after every 5 minutes of microwaving, but have never lost too much buffer volume since switching. And, the added bonus is that each container can hold up to 24 slides in a Tissue Tek plastic slide rack. Jan Shivers Univ. of Minnesota From shaumik.adhya <@t> ucl.ac.uk Thu Oct 21 10:42:57 2004 From: shaumik.adhya <@t> ucl.ac.uk (Shaumik Adhya) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] detection of HRP in FFPE sections- ?antibody Message-ID: <1098373377.4177d901a7095@www.webmail.ucl.ac.uk> Hi histonetters, I was wondering if any of you out there have experience of detection of HRP in formalin fixed, paraffin embedded tissue. I assume that the process of fixation and tissue processing will destroy the enzymatic activity of HRP, rendering the usual histochemical reagents useless. Are there antibodies out there that anyone's used? Thanks a lot for your help, Shaumik Adhya From kjnpeppa <@t> yahoo.com Thu Oct 21 10:50:03 2004 From: kjnpeppa <@t> yahoo.com (Barbara Rouse) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Problem with tissue processing in my VIP Message-ID: <20041021155003.22419.qmail@web51706.mail.yahoo.com> Hi everyone: We are having problems when we process our tissue specimens in the VIP. Whenever we process 80+ blocks (our VIP can hold 150 blocks), anything that is not a small biopsy does not process well (such as uteri, breast tissue, etc) even with specimens being held overnight or longer in formalin for fixation. We use zinc formalin, ethyl alcohol, Histo-Solv (xylene substitute - which we have been using for a long time, so I don't think that is the problem) and ParaPlast Xtra. Our times once machine starts processing: Zinc formalin, total of 2 hours 70% alcohol, 45 minutes 95% alcohol, total of 90 minutes 100% alcohol, total of 2-1/2 hours Histo-Solv, total of 2 hours Paraplast, total of 2 hours, 15 minutes Agitation is on and so is vacuum. Any ideas or help will be appreciated. Thanks in advance. Barbara kjnpeppa@yahoo.com --------------------------------- Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. From bwhitaker <@t> brownpathology.com Thu Oct 21 10:55:21 2004 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] re: microwave retrieval In-Reply-To: <008701c4b784$0c702a20$78065486@vdl220FAC> Message-ID: <000001c4b786$5f0c7890$3601a8c0@brownpathology.net> Be sure and fill the racks each time you do HIER. The heating will be more consistant if they always have the same amount of slides. (Use blanks over and over.) Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan Shivers Sent: Thursday, October 21, 2004 10:39 AM To: histonet Subject: [Histonet] re: microwave retrieval I previously used plastic coplin jars set into a tupperware-type square dish, but also had occasional evaporating/boiling down of buffer and subsequent uncovering/drying out of tissues (even though I'd check and add to the buffer level after every 5 minutes). Now I use Tissue-Tek plastic containers, the rectangular-shaped ones that are used in the deparaffinization/rehydration Tissue-Tek system. They hold more buffer volume, and it seems that less buffer is evaporated/boiled off. I still check buffer levels after every 5 minutes of microwaving, but have never lost too much buffer volume since switching. And, the added bonus is that each container can hold up to 24 slides in a Tissue Tek plastic slide rack. Jan Shivers Univ. of Minnesota _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DonnaWillis <@t> texashealth.org Thu Oct 21 11:05:45 2004 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] re: microwave retrieval Message-ID: Don't forget that BioGenex has a patent on HIER in a microwave. Donna Willis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonnie Whitaker Sent: Thursday, October 21, 2004 10:55 AM To: 'Jan Shivers'; 'histonet' Subject: RE: [Histonet] re: microwave retrieval Be sure and fill the racks each time you do HIER. The heating will be more consistant if they always have the same amount of slides. (Use blanks over and over.) Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan Shivers Sent: Thursday, October 21, 2004 10:39 AM To: histonet Subject: [Histonet] re: microwave retrieval I previously used plastic coplin jars set into a tupperware-type square dish, but also had occasional evaporating/boiling down of buffer and subsequent uncovering/drying out of tissues (even though I'd check and add to the buffer level after every 5 minutes). Now I use Tissue-Tek plastic containers, the rectangular-shaped ones that are used in the deparaffinization/rehydration Tissue-Tek system. They hold more buffer volume, and it seems that less buffer is evaporated/boiled off. I still check buffer levels after every 5 minutes of microwaving, but have never lost too much buffer volume since switching. And, the added bonus is that each container can hold up to 24 slides in a Tissue Tek plastic slide rack. Jan Shivers Univ. of Minnesota _______________________________________________ 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 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 are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From ploykasek <@t> phenopath.com Thu Oct 21 11:09:25 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:24:11 2005 Subject: [Histonet] Microwave retrieval In-Reply-To: Message-ID: Ed, I've done antigen retrieval many ways, and there are pros & cons to each. I believe consistency in what you do is the key to any antigen retrieval. I do not recommend microwaving with open containers - it is far too easy for there to be hot spots, loss of fluid, etc.... That being said, I do think it is possible to microwave retrieve in a closed container such as a microwave pressure cooker. Some of the keys to this technique are: use the same volume of liquid, the same # of slides (using blank slides if necessary) each & every time, monitoring with your microwave the time it takes to get up to pressure & how long you want your slides to remain under pressure, careful cooling down of slides. I?d be happy to provide the particulars of how we do it, if anyone is interested. In a microwave pressure cooker the fluid becomes ?super heated? - much hotter than in a steamer & you have the added pressure. We have found it improves staining with many difficult targets & nuclear antigens. I would note that I work at a reference lab, and our tissue is fixed & processed many different ways. If you can control the fixation & processing, and optimize it, I think a steamer works great & we use it for many antibodies. Again, consistency is the key. We preheat solutions before adding slides, only adding slides when the solution has reached 95 degrees C. To monitor the temperature we have thermometers with slender, metal probes that fit thru the holes in the steamer ( alternately, I?ve had my husband carefully enlarge the steamer lid holes to accommodate a standard thermometer). After adding slides, we start our retrieval time when the solution is back up to 95. We do the same retrieval time & cool down time each time. Well, I hope this helps you out. Feel free to contact me for more info. Good luck with your retrieval & staining. Patti Loykasek PhenoPath Laboratories Seattle, WA > Can anyone give me the pro's and con's of steamer/pressure (Decloaker) vs. > microwave retrieval ? > Our Doc wants us to change from the Decloaker to microwave. Is a household > microwave adequate or do we need a lab grade unit? > Does the microwave save time and improve turn around time? > > Thanks in advance > Ed Harris HT ASCP MT ABB > > > > This E-mail message (including attachments, if any) is intended for the use of > the individual or entity to which it is addressed and may contain information > that is privileged, proprietary, confidential and exempt from disclosure. If > you are not the intended recipient, you are notified that any dissemination, > distribution or copying of this communication is strictly prohibited. If you > have received this communication in error, please notify the sender and erase > this E-mail message immediately. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vitha <@t> mic.tamu.edu Thu Oct 21 11:23:39 2004 From: vitha <@t> mic.tamu.edu (Stanislav Vitha) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Re: Polyester wax -it does work for me too Message-ID: <6.1.2.0.0.20041021103030.0b851eb0@mic.tamu.edu> Hi Rebecca, I just joined the mailing list and found this old thread on Steedman's wax. I am glad Steedman's wax is working for you. I have used it extensively in last several years with good results for immunofluorescence labeling of many different epitopes in plant tissues (expanded leaves, meristems, root tips, flower buds, etc (I also embedded and sectioned mouse embryos and it worked fine as well). Using a standard, old-fashioned AO/Spencer rotary microtome, I am routinely able to get 5 micron sections if the air-conditioning works reasonably well, no need to use cold room or chill the block or the knife in the refrigerator (at least for the tissues I have worked with; it may help if you want even thinner sections). My specimens are usually rather small, the face of the sectioned block is generally about 5 x 4 mm, so for larger blocks it may be not as easy. I did not have the Section Transfer Station on my microtome, but several tricks helped: 1) During sectioning, I use a fine paint brush to lift the end of the ribbon and to hold the ribbon in the air so that it does not slide on the knife (it tends to stick to it if you do not) 2) For the 5 to 10 micron sections I use high sectioning speed (turn the wheel as fast as you can) and produce a ribbon up to about 30 cm long. I have cut 20 micron sections occasionally, using slower cutting speeds, but they tend to curl. I then use a second paint brush to lift and dislodge the other end of the ribbon from the knife edge. 3) I place the ribbon on a wooden board or a sheet of paper and using a razor blade, cut the ribbon into smaller pieces and place them on a dry, poly-L-lysine coated slide. - if you tilt the razor blade when cutting the piece, the short ribbon will stick to the edge well enough so that you can lift it and lay it on the microscope slide. 4) Add a drop of water to one end of each ribbon on the slide. The water will run under the ribbon and it will expand and stretch. Wick off excess water with filter paper. 5) Allow to dry at room temperature for few hours or overnight, then proceed with dewaxing and staining/immunostaining. More details and pictures on embedding and sectioning can be found in the following book chapter: Vitha, S., Balu?ka, F., Jasik, J., Volkmann, D., and Barlow, P., Steedman's wax for F-actin visualization, in Actin: a Dynamic Framework for Multiple Plant Cell Functions, C.J. Staiger, F. Balu?ka, D. Volkmann, and P. Barlow, Editors. 2000, Kluwer: Dordrecht, The Netherlands. p. 619-636. You can download the PDF from http://www.izmb.de/volkmann/pdfs/ (Book-Steedman's_wax.pdf) The disadvantage of the above method is that the ribbons sometimes do not expand 100%, or al least not all sections; for what I was doing it was not critical, I was more interested in cellular and subcellular immunostaining rather that looking at tissue and whole-organ morphology. For some of the immunolabeling, I have also used Superfrost Plus slides, but the section adherence is much worse than with the poly-L-lysine slides, especially if one needs to do antigen retrieval and/or lengthy incubations. (For some epitopes, I had to autoclave the tissue in high-pH buffer, and all sections were lost from the Superfrost Plus slides, while most remained on the lysin-coated ones). Good luck! Stan Vitha >From: Rebecca Nishi > >---------- > >I really appreciate everyones comments. It sounds like most everyone is not >that excited about it. > >I too read all of the literature from the 50s onward (there isnt too much), >and have tried Steedmans, and PEG-Disterate from Sigma. The procedure isnt >really too bad. > >As for my results so far, they are actually pretty good. I was hoping to get >some help with the fine tuning. I am confident I can get good 10 um >sections, using a rotary microtome (from Microm), I tried it with and >without a cool-cut adapter to keep the specimen cold (I think it was around >-10 to +4 C, I forgot), cold didn?t seem to help so we didn?t get that. But >what helped tremendously was the STS (Section transfer station). It is a >little waterfall and water bath, set to 30C. The sections slide perfectly >and flat down the waterfall, and into the bath, and are immediately mounted >on Superfrost Plus slides. I dry them on the slides for a day or 2 at room >temperature or 4C, then bake them at 30C -40C for about 15-30 minutes. > >I think a cooler room definitely helps. But I was cutting on the rotary >microtome, here in Southern California (mostly sunny, ~70-80F at the time) >and had no trouble when using the STS. We were having a lot of trouble >cryostatting recently (The past year), and thought we could get more stable >sections with this wax. I tried cutting the wax on the cryostat and with a >sliding microtome with little success. > >I was able to get nice 20uM sections as well, but with my current adhering >protocol, they came off the slides during staining. I plan to work on that >soon. > >I was unable to get 30 uM sections, they crumbled when hitting the knife. I >think 10 and thinner is optimal, but I would like to get 20 if possible. > >I can easily get ribbons that float on the pool. Once the section or ribbon >goes under the water, they are a little bit hard to handle, and unfold. You >have to put on a slide right away. Also, I don?t think you can store the >ribbons like other types of wax. I am only going to put 1 or 2 sections per >slide anyways because I plan to do stereology, and want multiple sets for >various stains. > >I think this method does have some potential, and I am particularly >interested in using this for injured areas, which tend to fall apart with >cryostat. I really have not found it that difficult to use, so far. > >Rebecca > >On 5/4/04 10:23 PM, "John Kiernan" wrote: > > > We tried Steedman's polyester wax in the early 1980s (Yes, > > after reading the early 1950s literature!). Powder rather > > than sections poured over the knife's edge. We gave up. > > > > In the light of recent (1990s) advances, are there any > > reasons for trying to master the lost art of sectioning > > polyester wax? This embedding medium was introduced > > shortly before the cryostat and long before antigen > > retrieval. In one of Steedman's procedures polyester wax > > was included in a one-step mixture with a Bouin-like > > fixative. > > After 24 hrs the liquid was cooled, and when it had set > > you could trim the block and (he said) produce ribbons > > of sections. > > > > Polyester wax reappeared in the 1970s, when it was widely > > thought that immunohistochemistry worked best after little > > or no fixation and avoidance of organic solvents, wax, > > heat etc. This didn't catch on despite being published in > > classy journals. > > > > The comments from Stephen.Eyres@sanofi-synthelabo.com > > (cited below) should help those who try to cut polyester > > wax. Clearly it's a difficult medium to handle. Are there > > any purposes for which it must be used? Dr. Stanislav Vitha vitha@mic.tamu.edu Microscopy and Imaging Center Texas A&M University BSBW 119 College Station, TX 77843-2257 tel: 979-845-1129 (main desk) tel: 979-845-1607 (direct link) fax: 979-847-8933 From xia <@t> oakland.edu Thu Oct 21 11:53:50 2004 From: xia <@t> oakland.edu (xia@oakland.edu) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Technician Position in Cartilage research Message-ID: Technician Position in Cartilage research A technician position will soon be available* in the Microscopic NMR Imaging (?MRI) Lab in Department of Physics, Oakland University. OU is located in suburban Rochester, Michigan, in north Oakland County, which boasts one of the most picturesque campuses in the country. We study articular cartilage using multidisciplinary techniques, including non-invasive ?MRI, polarized light microscopy and histology, Fourier-transform infrared imaging (FTIRI) system, and chemical and mechanical calibrations. MRI instrumentation consists of a Bruker AMX 300 NMR system with 7T wide-bore superconducting magnet, microimaging accessories, and a SGI workstation. Other major instruments in our lab include an EnduraTec ELF 3200 mechanical testing system, a Leica DM RXP polarized light microscope interfaced with a 12-bit CCD camera, a soon-to-be-installed FTIRI system, a biochemistry and histology lab, and a number of personal computers running Macintosh, UNIX, and Windows operating systems. The successful candidate should possess practical skills in the histology of cartilage or other connective tissue and a strong background in the routine operation of various modern instruments. Working knowledge of computers and Internet will be an asset. Specifically, we are looking for a highly motivated individual who will participate in the multidisciplinary research activities in our lab as part of a team. Additional responsibilities include routine-maintenance/trouble-shooting/upgrade/minor-repairs of existing instrument and computers, assistance in data analysis, training new students and other users, and other related duties as assigned. Handling live animal is not required. This is a full-time position funded for up to five years, renewable on a yearly basis given adequate performance. The position can also be tailored into part-time for a set of specific responsibilities. Interested individuals should send their CV, statement of previous experience, and the names, telephone numbers, and e-mail addresses of at least three references to: Dr. Yang Xia Associate Professor of Physics Dept of Physics, Oakland University, Rochester, MI 48309, USA Tel: 248-370-3420; Fax: 248-370-3408; E-mail: xia@oakland.edu Web: http://www.oakland.edu/~xia/XiaLab_index.html. * Pending final budgetary approval. OU is an equal opportunity employer. -- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Yang Xia, Ph. D. 276 Hannah Hall, Physics Department Oakland University, Rochester, MI 48309-4487, USA Tel: (248) 370-3420(office), 370-2403(NMR), 370-3402 & 370-4873 (labs) Fax: (248) 370-3408 Email: xia@oakland.edu Web: http://www.oakland.edu/~xia/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ From hymclab <@t> hyhc.com Thu Oct 21 12:08:32 2004 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Microwave retrieval Message-ID: Third Opinion! I did use a household microwave for years getting somewhat conistent results when doing them myself. Then we added more employees and the consistency thing went right out of the window. We got a rice steamer about a year ago and have had beautiful and consistent results ever since. I am very glad I did it!!! Dawn -----Original Message----- From: marjorie lehman [mailto:Marjorie.Lehman@unilever.com] Sent: Thursday, October 21, 2004 6:19 AM To: Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Microwave retrieval Here is a Second Opinion! I tried using a (Household) microwave for a long time and was not happy. After I got my Decloaking Chamber from Biocare I never looked back! Marge -----Original Message----- From: Amos Brooks [SMTP:amosbrooks@earthlink.net] Sent: Wednesday, October 20, 2004 8:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave retrieval Ed, I strongly recommend that you don't try to put a coplin jar of retrieval solution with slides in it in the microwave and expect any consistant results. What happens with this method is you get a spike of heat then a temperature drop. Very often there is boil over and your slides dry out. Ideally you should be looking for an even temperature for a defined length of time that can be well monitored. For this reason if you *must* use a microwave it should be a lab grade oven that has consistant wattage and temperature control. It would also be good to use one of the microwave pressure cookers to improve the consistency. (I think Biocare Medical has them ... the guys that sell Borg Decloaker). My ultimate recommendation is to stick to the tried and true waterbath or steamer method. It may take a bit longer but the consistancy is better and there is far less variables in the procedure. Just my opinion, You know what they say 'bout opinions, Amos Brooks At 01:00 PM 10/20/2004, you wrote: >Message: 10 >Date: Wed, 20 Oct 2004 11:30:26 -0400 >From: "Harris, Ed" >Subject: [Histonet] Microwave retrieval >To: "'histonet@lists.utsouthwestern.edu'" > >Message-ID: >Content-Type: text/plain; charset="iso-8859-1" > >Can anyone give me the pro's and con's of steamer/pressure (Decloaker) >vs. microwave retrieval ? Our Doc wants us to change from the Decloaker >to microwave. Is a household microwave adequate or do we need a lab >grade unit? Does the microwave save time and improve turn around time? > >Thanks in advance >Ed Harris HT ASCP MT ABB _______________________________________________ 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 From fjones <@t> namsa.com Thu Oct 21 13:03:52 2004 From: fjones <@t> namsa.com (Fawn Jones) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] MIDS Trichrome Brown Message-ID: <915E55B02E236E4D95258B181EEF6317075D44@namsams01.namsa.int> Does anybody know of a staining procedure for MIDS Trichrome Brown that they would be willing to share. I have a request to do this stain on paraffin sections as well as on hard plastic (polyester) sections en bloc. I have never heard of this stain and would appreciate any help on the matter. Thank you Fawn Jones From Luis.Chiriboga <@t> med.nyu.edu Thu Oct 21 11:41:46 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Microwave retrieval In-Reply-To: <83AACDB0810528418AA106F9AE9B7F7E278524@sjhaexc02.sjha.org> Message-ID: hi everyone Here's a slightly different view. I'm on my third household microwave in the past 6 years. I have used the microwave (occasionally MW pressure cooker) for all my HIER and have been able to get extremely reproducible results. If you look through the literature, you'll pretty much find that the source of heat is not important, it is the amount and time. That being said, the way I get around the "hotspot" and "drying" issues is to antigen retrieve in a large volume of retrieval buffer. I use 1000 ml beakers filled up to ~900 ml. I always start out with the same volume to be as consistent as possible. I have figured the amount of time it takes to get the volume boiling and just add that to my HIER time. periodically I check to make sure that the preheat time is still appropriate as the microwave generators do lose power over time. Obviously, this approach would not be applicable for those of you who by commercial grade retrieval buffers. We make up all are own stuff (20 L at a time) and go through it quite quickly. My lab is research primarily and when I showed one of our administrators the price of a laboratory grade microwave, he had a stroke. I guess when your poor (us research folks are on the bottom of the food chain) you have no choice and have to make do with what you got....... Hope this helps -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Weems, Joyce Sent: Thursday, October 21, 2004 8:15 AM To: marjorie lehman; Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Microwave retrieval We were fortunate enough to have some left over capital dollars and have purchased the new Biogenex Retreival Microwave. It is wonderful - saves lots of time and gives consistency in staining! The only thing we would change is to be have smaller containers for instances when we have just a few slides to retreive. But we think its a winner for those of you are looking to purchase it. Cheers, Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of marjorie lehman Sent: Thursday, October 21, 2004 7:19 AM To: Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Microwave retrieval Here is a Second Opinion! I tried using a (Household) microwave for a long time and was not happy. After I got my Decloaking Chamber from Biocare I never looked back! Marge -----Original Message----- From: Amos Brooks [SMTP:amosbrooks@earthlink.net] Sent: Wednesday, October 20, 2004 8:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave retrieval Ed, I strongly recommend that you don't try to put a coplin jar of retrieval solution with slides in it in the microwave and expect any consistant results. What happens with this method is you get a spike of heat then a temperature drop. Very often there is boil over and your slides dry out. Ideally you should be looking for an even temperature for a defined length of time that can be well monitored. For this reason if you *must* use a microwave it should be a lab grade oven that has consistant wattage and temperature control. It would also be good to use one of the microwave pressure cookers to improve the consistency. (I think Biocare Medical has them ... the guys that sell Borg Decloaker). My ultimate recommendation is to stick to the tried and true waterbath or steamer method. It may take a bit longer but the consistancy is better and there is far less variables in the procedure. Just my opinion, You know what they say 'bout opinions, Amos Brooks At 01:00 PM 10/20/2004, you wrote: >Message: 10 >Date: Wed, 20 Oct 2004 11:30:26 -0400 >From: "Harris, Ed" >Subject: [Histonet] Microwave retrieval >To: "'histonet@lists.utsouthwestern.edu'" > >Message-ID: >Content-Type: text/plain; charset="iso-8859-1" > >Can anyone give me the pro's and con's of steamer/pressure (Decloaker) vs. >microwave retrieval ? >Our Doc wants us to change from the Decloaker to microwave. Is a household >microwave adequate or do we need a lab grade unit? >Does the microwave save time and improve turn around time? > >Thanks in advance >Ed Harris HT ASCP MT ABB _______________________________________________ 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 Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From Luis.Chiriboga <@t> med.nyu.edu Thu Oct 21 08:44:54 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Microwave retrieval In-Reply-To: <83AACDB0810528418AA106F9AE9B7F7E278524@sjhaexc02.sjha.org> Message-ID: Hi everyone Here's a slightly different view. I'm on my third household microwave in the past 6 years. I have used the microwave (occasionally MW pressure cooker) for all my HIER and have been able to get extremely reproducible results. If you look through the literature, you'll pretty much find that the source of heat is not important, it is the amount and time. That being said, the way I get around the "hotspot" and "drying" issues is to antigen retrieve in a large volume of retrieval buffer. I use 1000 ml beakers filled up to ~900 ml. I always start out with the same volume to be as consistent as possible. I have figured the amount of time it takes to get the volume boiling and just add that to my HIER time. periodically I check to make sure that the preheat time is still appropriate as the microwave generators do lose power over time. Obviously, this approach would not be applicable for those of you who by commercial grade retrieval buffers. We make up all are own stuff (20 L at a time) and go through it quite quickly. My lab is research primarily and when I showed one of our administrators the price of a laboratory grade microwave, he had a stroke. I guess when your poor (us research folks are on the bottom of the food chain) you have no choice and have to make do with what you got....... Hope this helps -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Weems, Joyce Sent: Thursday, October 21, 2004 8:15 AM To: marjorie lehman; Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Microwave retrieval We were fortunate enough to have some left over capital dollars and have purchased the new Biogenex Retreival Microwave. It is wonderful - saves lots of time and gives consistency in staining! The only thing we would change is to be have smaller containers for instances when we have just a few slides to retreive. But we think its a winner for those of you are looking to purchase it. Cheers, Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of marjorie lehman Sent: Thursday, October 21, 2004 7:19 AM To: Amos Brooks; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Microwave retrieval Here is a Second Opinion! I tried using a (Household) microwave for a long time and was not happy. After I got my Decloaking Chamber from Biocare I never looked back! Marge -----Original Message----- From: Amos Brooks [SMTP:amosbrooks@earthlink.net] Sent: Wednesday, October 20, 2004 8:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave retrieval Ed, I strongly recommend that you don't try to put a coplin jar of retrieval solution with slides in it in the microwave and expect any consistant results. What happens with this method is you get a spike of heat then a temperature drop. Very often there is boil over and your slides dry out. Ideally you should be looking for an even temperature for a defined length of time that can be well monitored. For this reason if you *must* use a microwave it should be a lab grade oven that has consistant wattage and temperature control. It would also be good to use one of the microwave pressure cookers to improve the consistency. (I think Biocare Medical has them ... the guys that sell Borg Decloaker). My ultimate recommendation is to stick to the tried and true waterbath or steamer method. It may take a bit longer but the consistancy is better and there is far less variables in the procedure. Just my opinion, You know what they say 'bout opinions, Amos Brooks At 01:00 PM 10/20/2004, you wrote: >Message: 10 >Date: Wed, 20 Oct 2004 11:30:26 -0400 >From: "Harris, Ed" >Subject: [Histonet] Microwave retrieval >To: "'histonet@lists.utsouthwestern.edu'" > >Message-ID: >Content-Type: text/plain; charset="iso-8859-1" > >Can anyone give me the pro's and con's of steamer/pressure (Decloaker) vs. >microwave retrieval ? >Our Doc wants us to change from the Decloaker to microwave. Is a household >microwave adequate or do we need a lab grade unit? >Does the microwave save time and improve turn around time? > >Thanks in advance >Ed Harris HT ASCP MT ABB _______________________________________________ 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 Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From d.fuster <@t> ub.edu Thu Oct 21 13:33:12 2004 From: d.fuster <@t> ub.edu (Dolors Fuster) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Tendons Message-ID: <417800E8.7050702@ub.edu> Hello .... I've been asked to obtain histological sections from rabbit tendons with an acrylic fiber inside to see how the tissue incorpores the fiber. I can try to do it frozen or parafin embedding, although I think I can cut the acrylic fiber with no one of the methods. Any suggestions? Thanks in advance. From PatPatterson <@t> mhd.com Thu Oct 21 13:39:31 2004 From: PatPatterson <@t> mhd.com (Patterson, Pat) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Diff Quik stain Message-ID: <293C7C19EFF7D611AE1A0002A53F81140CB43AB5@omega.mhd.com> Our lab had been using silver stain for Helicobacter - can anyone recommend a Diff-Quik type stain kit for this? Thanks Pat Patterson Methodist Dallas *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. From c.m.vanderloos <@t> amc.uva.nl Thu Oct 21 13:56:25 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] RE: Anti-Dig options question for ISH Message-ID: <240f44241483.241483240f44@amc.uva.nl> Dear Julie, As far as I know those anti-dig antibodies are patented and produced by Roche. Likely other vendors of this product will get their stuff via Roche. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Juile Gorenstein Date Thu, 21 Oct 2004 05:48:32 -0700 (PDT) To Histonet Subject [Histonet] Anti-Dig options question for ISH Hello everyone, I was wondering, other than Roche, who else manufactures Anti-DIG antibodies. Does anyone have a favorite? (I am planning to use on Ventana Discovery) Thanks in advance! Julie Novartis Cambridge, MA From c.m.vanderloos <@t> amc.uva.nl Thu Oct 21 14:30:10 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] RE: NK1-C3 Message-ID: <23ca2d23ea79.23ea7923ca2d@amc.uva.nl> Dear Carmen, The antibody is named NKI-C3 instead of NK1-C3. It was once produced in Amsterdam by researchers at the Dutch Cancer Institute (Nederlands Kanker Instituut= NKI) and commercially available through Monosan (www.monosan.com) product number MON7003. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From carmen loiselle Date Thu, 21 Oct 2004 09:44:28 -0400 To histonet@pathology.swmed.edu Subject [Histonet] NK1-C3 Good morning all, Is anybody out there familiar with the antibody " NK1-C3" ?? It's a putative melanoma antigen. If so, can you tell me where to find it, which company etc... Thanks in advance From c.m.vanderloos <@t> amc.uva.nl Thu Oct 21 14:18:29 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] RE: detection of HRP in FFPE sections- ?antibody Message-ID: <370ba936bab4.36bab4370ba9@amc.uva.nl> Dear Shaumik, I guess that HRP was used as tracer in some kind of labeling experiment in an whole organ, animal or something, followed by fixation and embedding. As you know endogenous peroxidase activity in erythrocytes and neutrophils perfectly survives the whole embedding circus quite well (unfortunately to many of us). So would try this with your labeling HRP as well. There was a Dako antibody (rabbit) available long time ago. Also at www.biodesign.com. Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Shaumik Adhya Date Thu, 21 Oct 2004 16:42:57 +0100 To Histonet@lists.utsouthwestern.edu Subject [Histonet] detection of HRP in FFPE sections- ?antibody Hi histonetters, I was wondering if any of you out there have experience of detection of HRP in formalin fixed, paraffin embedded tissue. I assume that the process of fixation and tissue processing will destroy the enzymatic activity of HRP, rendering the usual histochemical reagents useless. Are there antibodies out there that anyone's used? Thanks a lot for your help, Shaumik Adhya From terribraud <@t> msn.com Thu Oct 21 14:27:35 2004 From: terribraud <@t> msn.com (TERRI BRAUD) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] RE: Microwave vs Steamer/Decloaker Message-ID: I read, with great interest, the discussion of microwave vs Steamer/decloaker. I would agree that achieving consistency in the process, from run to run, batch to batch, is the key. Personally, I've found that more difficult (but not impossible) to achieve in anything but a closed system, such as a steamer or pressure cooker. I started out using a microwave, then a microwave pressure cooker, and now use a digital, programmable, pressure cooker that offers me the desired consistency in temp, time, and pressure control. The reagents you use in that instrument and process are, of course, entirely up to you, and can make or break the successful labeling of an antigen site. What I do not find is a "one reagent fits all" that will sucessfully work for all tissues, all antibodies, and all fixatives and tissue processing, since all labs seem to have such great variability in that part of the process. I hope this helps. Terri Braud Surgical Pathology Manager Medical Laboratories University of Virgnina Health Systems Charlottesville, VA 22908 From darrenj <@t> medica.co.nz Thu Oct 21 14:47:51 2004 From: darrenj <@t> medica.co.nz (Darren James) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Diff Quik stain In-Reply-To: <293C7C19EFF7D611AE1A0002A53F81140CB43AB5@omega.mhd.com> Message-ID: <001001c4b7a6$d9c35d90$c864a8c0@medica.co.nz> Hi Pat, A large lab I worked in used Toluidine Blue set up on a Shandon Linistain for the large quantities of gastric and duodenal biopsies we received. This appeared to work very well(the Paths liked it)and it was a very simple regressive method. Darren -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patterson, Pat Sent: Friday, 22 October 2004 7:40 a.m. To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Diff Quik stain Our lab had been using silver stain for Helicobacter - can anyone recommend a Diff-Quik type stain kit for this? Thanks Pat Patterson Methodist Dallas *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Ben.Shelkowsky <@t> chomp.org Thu Oct 21 15:07:57 2004 From: Ben.Shelkowsky <@t> chomp.org (Shelkowsky, Ben) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Diff Quik stain Message-ID: <384DA2BD670CE34DA8D3B60F4ED7157F4D6150@exchsrvr.chomp.org> Our lab has been using the PROTOCOL Hema 3 setup. This is a quick 3 step stain that we stain for Helicobacter. We get it from Fisher Scientific. We are also trying a Richard Allan product which is also a 3 step procedure that is the same as the PROTOCOL product. It takes less than 1 minute total time to stain 1 slide at a time. I suppose with a little slide carrier you could stain up to 5 slides simultaneously. But we prefer to do it, one at a time. Hope this helps. Ben Shelkowsky HTL, (ASCP) ben.shelkowsky@chomp.org CHOMP Monterey, CA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Patterson, Pat Sent: Thu 10/21/2004 11:39 AM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Diff Quik stain Our lab had been using silver stain for Helicobacter - can anyone recommend a Diff-Quik type stain kit for this? Thanks Pat Patterson Methodist Dallas *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This is a transmission from Community Hospital of the Monterey Peninsula. This message and any attached documents may be confidential and contain information protected by state and federal medical privacy statutes. They are intended only for the use of the addressee. If you are not the intended recipient, any disclosure, copying, or distribution of this information is strictly prohibited. If you received this transmission in error, please accept our apologies and notify the sender. Thank you. From betsy.fink <@t> uky.edu Thu Oct 21 14:51:50 2004 From: betsy.fink <@t> uky.edu (Betsy Fink) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] muscle staining Message-ID: <00a501c4b7a7$68476f70$a317a380@universi6b29b2> I was wondering if anyone is familiar with staining skeletal muscle grossly with nitrotetrazolium blue, to determine viable cells after reperfusion injury? We have done this successfully a few years back with pig rectus muscle. We now want to use this method with the rat gracilis muscle. Any thoughts or comments would be appreciated. From Jackie.O'Connor <@t> abbott.com Thu Oct 21 15:34:52 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Tendons Message-ID: I've done this with gore-tex grafts in carotids - in paraffin sections routinely processed. They were pretty nice - I don't know if the acrylic would hold up to xylene, however. So, my answer is pretty much useless - - Dolors Fuster Sent by: histonet-bounces@lists.utsouthwestern.edu 10/21/2004 01:33 PM To: HISTONET NOU cc: Subject: [Histonet] Tendons Hello .... I've been asked to obtain histological sections from rabbit tendons with an acrylic fiber inside to see how the tissue incorpores the fiber. I can try to do it frozen or parafin embedding, although I think I can cut the acrylic fiber with no one of the methods. Any suggestions? Thanks in advance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Sandra.Etheridge <@t> gems8.gov.bc.ca Thu Oct 21 16:04:47 2004 From: Sandra.Etheridge <@t> gems8.gov.bc.ca (Etheridge, Sandra AGF:EX) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Giardia Primary Antibody Message-ID: <424FAC08A8CDFE4BADD6497AE75B5E2F0E0A8792@atlas.gov.bc.ca> Hello everyone, We are looking for a primary antibody for Giardia. Is anyone out there in the veterinary research community aware of anyone who will supply us with some?? I have checked out numerous websites and found one lab who tests for it in Michigan (from the South Dakota State IHC web page), but have not heard back from them. Any info is, as always, appreciated. Thanks! Sandra Etheridge BC Ministry of Agriculture, Food & Fisheries Animal Health Center, Histology Abbotsford, BC Canada From gentras <@t> vetmed.auburn.edu Thu Oct 21 16:11:14 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] relin or relen (?sp) Ab Message-ID: <6.0.1.1.0.20041021160613.01af7438@mailhost.vetmed.auburn.edu> Hello, is anyone familiar with an IHC protocol using relin/relen ab on formalin fixed, paraffin embedded brain tissue? If so will you please share it with me? Thanks! Atoska From AnthonyH <@t> chw.edu.au Thu Oct 21 18:41:46 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] FETAL BRAIN FIXATION Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E0DC@simba.kids> We have also used a modified Bouin's Fixative for our fetal brains. Bryan do you have a reference for the modified Brasil's Fixative? The Brasil's fixative that we have used for liver biopsies where a metabolic disease is suspected is as follows: Absolute alcohol 1650ml Formalin 600ml Trichloracetic acid 7.5g Picric acid 10g It has been suggested as a good fixative for glycogen, though I can't find a reference for this. The above solution has been used in my lab for years, before I arrived here, hence my confusion. In fact one of our recent papers used this fixative and it had to be published with out a reference, so if anyone can shed some light I would be most appreciative: V-M. Mangan, V. Farago, M. Kelly, and A. F. Henwood (2002) " An Amylase Reagent with a Long Shelf Life for the Removal of Glycogen from Tissue Sections" J Histotechnol. 25(3): 153-4. You will note that our Brasil's fixative contains picric acid. The nomenclature seems very confused! Any insights? Regards, Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Bryan Hewlett [mailto:bhewlett@cogeco.ca] Sent: Thursday, 21 October 2004 11:33 PM To: lpwenk@sbcglobal.net; Histonet Subject: Re: [Histonet] FETAL BRAIN FIXATION Peggy, We faced the same issue a number of years ago. Our solution was to use a modified Brasil fixative. The pediatric pathologist was delighted with the results! The formula follows; Modified Brasil Fluid. Ethanol............................................. 700 mL 40% w/v Formaldehyde.....................100 mL Glacial Acetic acid..............................50 mL add D.water to make up to.................1000 mL Fetal brains are left to fix for 1-3 weeks, rinsed in 70% ETOH, then sliced and photographed. Selected blocks are processed using the same reagents and times as for NBF fixed material. Hope this helps, Bryan ----- Original Message ----- From: To: "Histonet" Sent: Thursday, October 21, 2004 4:56 AM Subject: [Histonet] FETAL BRAIN FIXATION > Anyone have a good fixative for the very soft fetal brains? > > Our neuropathologist does not want to use Bouin, as the tissue turns yellow > and photographs do not look the way they are supposed to. > > Another one of our pathologists remembers using an alcohol-acetone-formalin > mixture years ago as a resident in another institution, but doesn't know the > percentage of any of the reagents. > > I've tried several search engines and PubMed, and can't find anything. > > Thanks. > > Peggy A. Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > > _______________________________________________ > 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 ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From emry <@t> u.washington.edu Thu Oct 21 19:31:23 2004 From: emry <@t> u.washington.edu (P. Emry) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] stains..fiber..cont.tissue Message-ID: Hi, I need the simplest stain for fibers or connective tissues for decalcified bone sutures. I am doing an H & E also, but I would like to dazzle the boss without working very hard. Thanks, Trisha From rockbeki <@t> ufl.edu Thu Oct 21 21:29:45 2004 From: rockbeki <@t> ufl.edu (SMITH,REBEKAH FELICIA) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Microwave retrieval Message-ID: <52842769.1098412185761.JavaMail.osg@osgjas01.cns.ufl.edu> Being myself a poor research tech, I also use the household microwave method. I stick my slides in coplin jars filled with Tris buffer with the tops off then stick them in another container and microwave for 45sec twice, changing the water in between. It's been working for me. SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" From doscwk <@t> nus.edu.sg Thu Oct 21 22:31:24 2004 From: doscwk <@t> nus.edu.sg (Chan Wai Kam) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Fixative for tunel procedure Message-ID: Hi Netters, I'm processing rabbit spine specimens for normal histology and also the tunel procedure for paraffin sections, and I'm using 10% formol calcium as fixative. The procedures for the tunel procedure which I referred to recommend using 4% paraformaldehye as fixative. Does using formol calcium as fixative make any difference to the end result for the tunel procedure? I would appreciate your advice. Thanks Julee Chan Dept of Orthopaedic Surgery National University of Singapore From bhewlett <@t> cogeco.ca Thu Oct 21 22:57:36 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] FETAL BRAIN FIXATION References: <1CF2E2E5BB36D5119E7A0008C791F3741269E0DC@simba.kids> Message-ID: <001601c4b7eb$44de6680$6400a8c0@mainbox> Tony, You are not alone in being confused by the nomenclature of fixatives. I share your confusion and so do most authors of textbooks!!! Anyone making a slight modification to a fixative mixture seems to have put his name on it!!! There are numerous examples in the texts. There can be little doubt that Clarke (1851) was the originator of using a combination of alcohol and acetic acid for fixation, yet many authors attribute Carnoy (1887). Indeed, Carnoy did use it (Carnoy A), but only his contribution of adding chloroform to the mixture (Carnoy B) should be acknowledged (see Baker and Lillie). (Reference: Clarke J L.(1851) Researches into the structure of the spinal cord, Phil. Trans. R.S. 141: 601-622) Following the introduction of formaldehyde (Blum 1893) a great number of people utilized it in various combinations with other fixatives. Most notably Bouin (1897), who combined it with picric and acetic acids the result of which is still in wide use today. Brasil (1905) also experimented with added formaldehyde in several mixtures.He used the combination of formaldehyde and Clarke's fluid for general histological work (the one I gave) and an alcoholic version of Bouin's fluid for protozoa. It is the latter which is commonly called Brasil's fluid!! (Reference: Brasil, L.(1905) Nouvelles recherches sur la reproduction des gregarines monocystidees, Arch. Zool. Exp. 4: 69-99) Brasil's combination of formaldehyde, acetic acid and alcohol as a fixative mixture is known by many names. Lillie lists Tellyesniczky, Fekete, Opie & Lavin, Bodian and Lillie's FAA as the same fixative with only minor variations in the individual component concentrations! None the less Brasil was the originator. Incidentally, I can find no reference to the substitution of trichloracetic for the acetic acid in the original Brasil's fluid you are using! The formula I have gives: 80% alcohol 1500 mL Formalin 600 mL Glacial acetic acid 150 mL Picric acid 10 g Your formula is no doubt another of those minor modifications that actually went unnamed!! You will find a reference to the so-called good fixation of glycogen by Brasil and similar fixatives in Kiernan's textbook under Gendre's fluid, yet another name for alcoholic Bouin!!!!! Yours in confusion, Bryan ----- Original Message ----- From: "Tony Henwood" To: "'Bryan Hewlett'" ; ; "Histonet" Sent: Thursday, October 21, 2004 7:41 PM Subject: RE: [Histonet] FETAL BRAIN FIXATION > > We have also used a modified Bouin's Fixative for our fetal brains. > Bryan do you have a reference for the modified Brasil's Fixative? > The Brasil's fixative that we have used for liver biopsies where a metabolic > disease is suspected is as follows: > > Absolute alcohol 1650ml > Formalin 600ml > Trichloracetic acid 7.5g > Picric acid 10g > > It has been suggested as a good fixative for glycogen, though I can't find a > reference for this. The above solution has been used in my lab for years, > before I arrived here, hence my confusion. In fact one of our recent papers > used this fixative and it had to be published with out a reference, so if > anyone can shed some light I would be most appreciative: > > V-M. Mangan, V. Farago, M. Kelly, and A. F. Henwood (2002) " An Amylase > Reagent with a Long Shelf Life for the Removal of Glycogen from Tissue > Sections" J Histotechnol. 25(3): 153-4. > > You will note that our Brasil's fixative contains picric acid. > The nomenclature seems very confused! > Any insights? > > Regards, > > Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) > Laboratory Manager > The Children's Hospital at Westmead, > Locked Bag 4001, Westmead, 2145, AUSTRALIA. > Tel: 612 9845 3306 > Fax: 612 9845 3318 > > -----Original Message----- > From: Bryan Hewlett [mailto:bhewlett@cogeco.ca] > Sent: Thursday, 21 October 2004 11:33 PM > To: lpwenk@sbcglobal.net; Histonet > Subject: Re: [Histonet] FETAL BRAIN FIXATION > > > Peggy, > > We faced the same issue a number of years ago. > Our solution was to use a modified Brasil fixative. > The pediatric pathologist was delighted with the results! > The formula follows; > > Modified Brasil Fluid. > > Ethanol............................................. 700 mL > 40% w/v Formaldehyde.....................100 mL > Glacial Acetic acid..............................50 mL > add D.water to make up to.................1000 mL > > Fetal brains are left to fix for 1-3 weeks, rinsed in 70% ETOH, then sliced > and photographed. > Selected blocks are processed using the same reagents and times as for NBF > fixed material. > > Hope this helps, > > Bryan > > ----- Original Message ----- > From: > To: "Histonet" > Sent: Thursday, October 21, 2004 4:56 AM > Subject: [Histonet] FETAL BRAIN FIXATION > > > > Anyone have a good fixative for the very soft fetal brains? > > > > Our neuropathologist does not want to use Bouin, as the tissue turns > yellow > > and photographs do not look the way they are supposed to. > > > > Another one of our pathologists remembers using an > alcohol-acetone-formalin > > mixture years ago as a resident in another institution, but doesn't know > the > > percentage of any of the reagents. > > > > I've tried several search engines and PubMed, and can't find anything. > > > > Thanks. > > > > Peggy A. Wenk, HTL(ASCP)SLS > > William Beaumont Hospital > > Royal Oak, MI 48073 > > > > > > _______________________________________________ > > 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 > > > ********************************************************************** > This email and any files transmitted with it are confidential and > intended solely for the use of the individual or entity to whom they > are addressed. If you are not the intended recipient, please > delete it and notify the sender. > > Views expressed in this message and any attachments are those > of the individual sender, and are not necessarily the views of The > Children's Hospital at Westmead > > This footnote also confirms that this email message has been > virus scanned and although no computer viruses were detected, > the Childrens Hospital at Westmead accepts no liability for any > consequential damage resulting from email containing computer > viruses. > ********************************************************************** > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From AnthonyH <@t> chw.edu.au Thu Oct 21 23:30:48 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] FETAL BRAIN FIXATION Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E0E2@simba.kids> BRILLIANT Thanks Bryan. At last I have some references to the conundrum that is the Brasil's fixatives ( or should I say Bouin's or Clarkes, or Carnoy's or Gendre's.... Or Henwood's!! Why Not?). (Tongue in cheek!!) Regards, Tony -----Original Message----- From: Bryan Hewlett [mailto:bhewlett@cogeco.ca] Sent: Friday, 22 October 2004 1:58 PM To: Tony Henwood; lpwenk@sbcglobal.net; Histonet Subject: RE: Re: [Histonet] FETAL BRAIN FIXATION Tony, You are not alone in being confused by the nomenclature of fixatives. I share your confusion and so do most authors of textbooks!!! Anyone making a slight modification to a fixative mixture seems to have put his name on it!!! There are numerous examples in the texts. There can be little doubt that Clarke (1851) was the originator of using a combination of alcohol and acetic acid for fixation, yet many authors attribute Carnoy (1887). Indeed, Carnoy did use it (Carnoy A), but only his contribution of adding chloroform to the mixture (Carnoy B) should be acknowledged (see Baker and Lillie). (Reference: Clarke J L.(1851) Researches into the structure of the spinal cord, Phil. Trans. R.S. 141: 601-622) Following the introduction of formaldehyde (Blum 1893) a great number of people utilized it in various combinations with other fixatives. Most notably Bouin (1897), who combined it with picric and acetic acids the result of which is still in wide use today. Brasil (1905) also experimented with added formaldehyde in several mixtures.He used the combination of formaldehyde and Clarke's fluid for general histological work (the one I gave) and an alcoholic version of Bouin's fluid for protozoa. It is the latter which is commonly called Brasil's fluid!! (Reference: Brasil, L.(1905) Nouvelles recherches sur la reproduction des gregarines monocystidees, Arch. Zool. Exp. 4: 69-99) Brasil's combination of formaldehyde, acetic acid and alcohol as a fixative mixture is known by many names. Lillie lists Tellyesniczky, Fekete, Opie & Lavin, Bodian and Lillie's FAA as the same fixative with only minor variations in the individual component concentrations! None the less Brasil was the originator. Incidentally, I can find no reference to the substitution of trichloracetic for the acetic acid in the original Brasil's fluid you are using! The formula I have gives: 80% alcohol 1500 mL Formalin 600 mL Glacial acetic acid 150 mL Picric acid 10 g Your formula is no doubt another of those minor modifications that actually went unnamed!! You will find a reference to the so-called good fixation of glycogen by Brasil and similar fixatives in Kiernan's textbook under Gendre's fluid, yet another name for alcoholic Bouin!!!!! Yours in confusion, Bryan ----- Original Message ----- From: "Tony Henwood" To: "'Bryan Hewlett'" ; ; "Histonet" Sent: Thursday, October 21, 2004 7:41 PM Subject: RE: [Histonet] FETAL BRAIN FIXATION > > We have also used a modified Bouin's Fixative for our fetal brains. > Bryan do you have a reference for the modified Brasil's Fixative? > The Brasil's fixative that we have used for liver biopsies where a metabolic > disease is suspected is as follows: > > Absolute alcohol 1650ml > Formalin 600ml > Trichloracetic acid 7.5g > Picric acid 10g > > It has been suggested as a good fixative for glycogen, though I can't find a > reference for this. The above solution has been used in my lab for years, > before I arrived here, hence my confusion. In fact one of our recent papers > used this fixative and it had to be published with out a reference, so if > anyone can shed some light I would be most appreciative: > > V-M. Mangan, V. Farago, M. Kelly, and A. F. Henwood (2002) " An Amylase > Reagent with a Long Shelf Life for the Removal of Glycogen from Tissue > Sections" J Histotechnol. 25(3): 153-4. > > You will note that our Brasil's fixative contains picric acid. > The nomenclature seems very confused! > Any insights? > > Regards, > > Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) > Laboratory Manager > The Children's Hospital at Westmead, > Locked Bag 4001, Westmead, 2145, AUSTRALIA. > Tel: 612 9845 3306 > Fax: 612 9845 3318 > > -----Original Message----- > From: Bryan Hewlett [mailto:bhewlett@cogeco.ca] > Sent: Thursday, 21 October 2004 11:33 PM > To: lpwenk@sbcglobal.net; Histonet > Subject: Re: [Histonet] FETAL BRAIN FIXATION > > > Peggy, > > We faced the same issue a number of years ago. > Our solution was to use a modified Brasil fixative. > The pediatric pathologist was delighted with the results! > The formula follows; > > Modified Brasil Fluid. > > Ethanol............................................. 700 mL > 40% w/v Formaldehyde.....................100 mL > Glacial Acetic acid..............................50 mL > add D.water to make up to.................1000 mL > > Fetal brains are left to fix for 1-3 weeks, rinsed in 70% ETOH, then sliced > and photographed. > Selected blocks are processed using the same reagents and times as for NBF > fixed material. > > Hope this helps, > > Bryan > > ----- Original Message ----- > From: > To: "Histonet" > Sent: Thursday, October 21, 2004 4:56 AM > Subject: [Histonet] FETAL BRAIN FIXATION > > > > Anyone have a good fixative for the very soft fetal brains? > > > > Our neuropathologist does not want to use Bouin, as the tissue turns > yellow > > and photographs do not look the way they are supposed to. > > > > Another one of our pathologists remembers using an > alcohol-acetone-formalin > > mixture years ago as a resident in another institution, but doesn't know > the > > percentage of any of the reagents. > > > > I've tried several search engines and PubMed, and can't find anything. > > > > Thanks. > > > > Peggy A. Wenk, HTL(ASCP)SLS > > William Beaumont Hospital > > Royal Oak, MI 48073 > > > > > > _______________________________________________ > > 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 > > > ********************************************************************** > This email and any files transmitted with it are confidential and > intended solely for the use of the individual or entity to whom they > are addressed. If you are not the intended recipient, please > delete it and notify the sender. > > Views expressed in this message and any attachments are those > of the individual sender, and are not necessarily the views of The > Children's Hospital at Westmead > > This footnote also confirms that this email message has been > virus scanned and although no computer viruses were detected, > the Childrens Hospital at Westmead accepts no liability for any > consequential damage resulting from email containing computer > viruses. > ********************************************************************** > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Marion.Hiles <@t> north-bristol.swest.nhs.uk Fri Oct 22 06:16:26 2004 From: Marion.Hiles <@t> north-bristol.swest.nhs.uk (Marion Hiles) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Problem with tissue processing in my VIP Message-ID: <2EE924DF60902943AC6E2EF35155451F19CD3C@nbfexch03.north-bristol.nhs> Ditch the zinc formalin and use 10% neutral buffered formalin and it seems the processing times are a bit short. Try extending the clearing and wax stages. Bob Quilty Neuropath Frenchay Hospital Bristol UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Barbara Rouse Sent: 21 October 2004 16:50 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Problem with tissue processing in my VIP Hi everyone: We are having problems when we process our tissue specimens in the VIP. Whenever we process 80+ blocks (our VIP can hold 150 blocks), anything that is not a small biopsy does not process well (such as uteri, breast tissue, etc) even with specimens being held overnight or longer in formalin for fixation. We use zinc formalin, ethyl alcohol, Histo-Solv (xylene substitute - which we have been using for a long time, so I don't think that is the problem) and ParaPlast Xtra. Our times once machine starts processing: Zinc formalin, total of 2 hours 70% alcohol, 45 minutes 95% alcohol, total of 90 minutes 100% alcohol, total of 2-1/2 hours Histo-Solv, total of 2 hours Paraplast, total of 2 hours, 15 minutes Agitation is on and so is vacuum. Any ideas or help will be appreciated. Thanks in advance. Barbara kjnpeppa@yahoo.com --------------------------------- Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet DISCLAIMER: The information in this message is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorised. If you are not the intended recipient, any disclosure, copying, or distribution of the message, or any action or omission taken by you in reliance on it, is prohibited and may be unlawful. Please immediately contact the sender if you have received this message in error. Thank you. From dobbin <@t> upei.ca Fri Oct 22 06:47:07 2004 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:24:12 2005 Subject: (Fwd) Re: [Histonet] Giardia Primary Antibody Message-ID: <4178C90A.30772.E384F@localhost> ------- Forwarded message follows ------- From: Greg Dobbin To: "Etheridge, Sandra AGF:EX" Subject: Re: [Histonet] Giardia Primary Antibody Date sent: Fri, 22 Oct 2004 08:46:21 ADT Hi Sandra, Here at AVC we just had a young parasitologist leave for a job in Perth, Australia, his name is Dr. Ryan O'Hanley and giardia is his thing. He was using an Ab for IFATs. He bought his Ab's from Waterbourne Inc. in New Orleans. He ordered "Giardia/A/Glow" Cat # A300. Sounds like a FITC conjugated Ab, but it's a start. Have a nice weekend. Greg From: "Etheridge, Sandra AGF:EX" To: " (histonet@lists.utsouthwestern.edu)" Date sent: Thu, 21 Oct 2004 14:04:47 -0700 Subject: [Histonet] Giardia Primary Antibody > Hello everyone, > > We are looking for a primary antibody for Giardia. Is anyone out there in > the veterinary research community aware of anyone who will supply us with > some?? I have checked out numerous websites and found one lab who tests for > it in Michigan (from the South Dakota State IHC web page), but have not > heard back from them. > > Any info is, as always, appreciated. > > Thanks! > > Sandra Etheridge > > BC Ministry of Agriculture, Food & Fisheries > Animal Health Center, Histology > Abbotsford, BC > Canada > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------- End of forwarded message ------- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Pathology Lab Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Happiness is a journey, not a destination. From mark.lewis <@t> thermo.com Fri Oct 22 07:46:48 2004 From: mark.lewis <@t> thermo.com (mark.lewis@thermo.com) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Microwave retrieval Message-ID: Patty, I think you hit the nail on the head when you said "I believe consistency in what you do is the key to any antigen retrieval." I believe it is important for all aspects of the Histo lab. It makes trouble-shooting problems much easier if the work is all done in a consistent manner. Have a nice day Best regards, Mark Mark Lewis Product Specialist Anatomical Pathology Clinical Diagnostics Thermo Electron Corporation (412) 747-4013 (412) 788-1097 E-mail: mark.lewis@thermo.com Patti Loykasek To: histonet Sent by: cc: histonet-bounces@lists.utsouth Subject: Re: [Histonet] Microwave retrieval western.edu 10/21/2004 12:09 PM Ed, I've done antigen retrieval many ways, and there are pros & cons to each. I believe consistency in what you do is the key to any antigen retrieval. I do not recommend microwaving with open containers - it is far too easy for there to be hot spots, loss of fluid, etc.... That being said, I do think it is possible to microwave retrieve in a closed container such as a microwave pressure cooker. Some of the keys to this technique are: use the same volume of liquid, the same # of slides (using blank slides if necessary) each & every time, monitoring with your microwave the time it takes to get up to pressure & how long you want your slides to remain under pressure, careful cooling down of slides. I?d be happy to provide the particulars of how we do it, if anyone is interested. In a microwave pressure cooker the fluid becomes ?super heated? - much hotter than in a steamer & you have the added pressure. We have found it improves staining with many difficult targets & nuclear antigens. I would note that I work at a reference lab, and our tissue is fixed & processed many different ways. If you can control the fixation & processing, and optimize it, I think a steamer works great & we use it for many antibodies. Again, consistency is the key. We preheat solutions before adding slides, only adding slides when the solution has reached 95 degrees C. To monitor the temperature we have thermometers with slender, metal probes that fit thru the holes in the steamer ( alternately, I?ve had my husband carefully enlarge the steamer lid holes to accommodate a standard thermometer). After adding slides, we start our retrieval time when the solution is back up to 95. We do the same retrieval time & cool down time each time. Well, I hope this helps you out. Feel free to contact me for more info. Good luck with your retrieval & staining. Patti Loykasek PhenoPath Laboratories Seattle, WA > Can anyone give me the pro's and con's of steamer/pressure (Decloaker) vs. > microwave retrieval ? > Our Doc wants us to change from the Decloaker to microwave. Is a household > microwave adequate or do we need a lab grade unit? > Does the microwave save time and improve turn around time? > > Thanks in advance > Ed Harris HT ASCP MT ABB > > > > This E-mail message (including attachments, if any) is intended for the use of > the individual or entity to which it is addressed and may contain information > that is privileged, proprietary, confidential and exempt from disclosure. If > you are not the intended recipient, you are notified that any dissemination, > distribution or copying of this communication is strictly prohibited. If you > have received this communication in error, please notify the sender and erase > this E-mail message immediately. > > _______________________________________________ > 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 From Catherine.Goeden <@t> med.va.gov Fri Oct 22 08:42:01 2004 From: Catherine.Goeden <@t> med.va.gov (Goeden, Catherine) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] CAP question Message-ID: <597FCC5997E8504EAA6814E6C9ED52298FAD6A@VHASUXEXC1> We are getting ready for a CAP inspection and I have a question about how other techs are handling a question regarding waterbaths: ANP.23350 Are they clean and well-maintained, and is there a policy for preventing cross-contamination of paraffin sections in the bath? I have a procedure for cutting the sections and cleaning water baths but I guess I am not specifically using the word cross-contamination. Would appreciate how other labs are "wording" this procedure. Thanks in advance!! From amosbrooks <@t> earthlink.net Fri Oct 22 09:48:40 2004 From: amosbrooks <@t> earthlink.net (amosbrooks@earthlink.net) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Microwave retrieval Message-ID: <10909363.1098456520591.JavaMail.root@skeeter.psp.pas.earthlink.net> Hi, Just to point out a steamer is cheaper than a microwave. So for a lab on a budget the steamer is even better. You should give it a whirl some time. You may find some improved consistancy. Of course if you are content with what you have I stand by something I always say: All labs are different, What works fine in one lab may cause problems in another. If it aint broke don't fix it. Amos -----Original Message----- From: "SMITH,REBEKAH FELICIA" Sent: Oct 21, 2004 10:29 PM To: Luis Chiriboga , "Weems, Joyce" , marjorie lehman , Amos Brooks , histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Microwave retrieval Being myself a poor research tech, I also use the household microwave method. I stick my slides in coplin jars filled with Tris buffer with the tops off then stick them in another container and microwave for 45sec twice, changing the water in between. It's been working for me. SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" From tobetrice <@t> yahoo.com Fri Oct 22 11:28:14 2004 From: tobetrice <@t> yahoo.com (Patrice McCall) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] reference for plastics Message-ID: <20041022162814.84913.qmail@web54504.mail.yahoo.com> Does anyone know a good reference book or other material to use for learning to do plastics in histology? __________________________________ Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. http://promotions.yahoo.com/new_mail From portera203 <@t> yahoo.com Fri Oct 22 12:08:42 2004 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Problem with tissue processing in my VIP In-Reply-To: <2EE924DF60902943AC6E2EF35155451F19CD3C@nbfexch03.north-bristol.nhs> Message-ID: <20041022170842.97878.qmail@web40914.mail.yahoo.com> Not that it is always the machine, but you might want to have a service call just to be sure that there are no problems with the processor. At one time we had a problem with the diaphragm being cracked, but not enough to cause an alarm for vacuum or pressure. We were getting some cross contamination of reagents which caused "mushy" uninfiltrated pieces of some tissue types. If modifying your processing protocol does not help the situation and you still have problems, don't assume that it is something you are not doing right....good luck solving the problem. Marion Hiles wrote:Ditch the zinc formalin and use 10% neutral buffered formalin and it seems the processing times are a bit short. Try extending the clearing and wax stages. Bob Quilty Neuropath Frenchay Hospital Bristol UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Barbara Rouse Sent: 21 October 2004 16:50 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Problem with tissue processing in my VIP Hi everyone: We are having problems when we process our tissue specimens in the VIP. Whenever we process 80+ blocks (our VIP can hold 150 blocks), anything that is not a small biopsy does not process well (such as uteri, breast tissue, etc) even with specimens being held overnight or longer in formalin for fixation. We use zinc formalin, ethyl alcohol, Histo-Solv (xylene substitute - which we have been using for a long time, so I don't think that is the problem) and ParaPlast Xtra. Our times once machine starts processing: Zinc formalin, total of 2 hours 70% alcohol, 45 minutes 95% alcohol, total of 90 minutes 100% alcohol, total of 2-1/2 hours Histo-Solv, total of 2 hours Paraplast, total of 2 hours, 15 minutes Agitation is on and so is vacuum. Any ideas or help will be appreciated. Thanks in advance. Barbara kjnpeppa@yahoo.com --------------------------------- Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet DISCLAIMER: The information in this message is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorised. If you are not the intended recipient, any disclosure, copying, or distribution of the message, or any action or omission taken by you in reliance on it, is prohibited and may be unlawful. Please immediately contact the sender if you have received this message in error. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Amy S.Porter, HT(ASCP) Michigan State University Department of Physiology Division of Human Pathology College of Human Medicine portera203@yahoo.com --------------------------------- Do you Yahoo!? Yahoo! Mail – CNET Editors' Choice 2004. Tell them what you think. a From Rachael_Emerson <@t> URMC.Rochester.edu Fri Oct 22 13:27:13 2004 From: Rachael_Emerson <@t> URMC.Rochester.edu (Emerson, Rachael) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] RE: Histonet Digest, Vol 11, Issue 30 Message-ID: Hello. Does anyone have any experience working with CD41, GPV, and GPIBbeta antibodies? I am looking for protocols to use them for immunohistochemistry on mouse embryos fixed in paraformaldehyde. THANK YOU! Rachael L. Emerson Center for Human Genetics and Molecular Pediatric Diseases University of Rochester Medical Center 575 Elmwood Avenue MRBX 1.11301 Rochester, NY 14642 Tel (585) 275-5073 Fax (585) 276-0232 > ---------- > From: histonet-request@lists.utsouthwestern.edu > Reply To: histonet@lists.utsouthwestern.edu > Sent: Friday, October 22, 2004 1:03 PM > To: histonet@lists.utsouthwestern.edu > Subject: Histonet Digest, Vol 11, Issue 30 > > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > > > Today's Topics: > > 1. (Fwd) Re: [Histonet] Giardia Primary Antibody (Greg Dobbin) > 2. Re: Microwave retrieval (mark.lewis@thermo.com) > 3. CAP question (Goeden, Catherine) > 4. RE: Microwave retrieval (amosbrooks@earthlink.net) > 5. reference for plastics (Patrice McCall) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Fri, 22 Oct 2004 08:47:07 ADT > From: "Greg Dobbin" > Subject: (Fwd) Re: [Histonet] Giardia Primary Antibody > To: histonet@lists.utsouthwestern.edu > Message-ID: <4178C90A.30772.E384F@localhost> > Content-Type: text/plain; charset=US-ASCII > > > ------- Forwarded message follows ------- > From: Greg Dobbin > To: "Etheridge, Sandra AGF:EX" > > Subject: Re: [Histonet] Giardia Primary Antibody > Date sent: Fri, 22 Oct 2004 08:46:21 ADT > > Hi Sandra, > Here at AVC we just had a young parasitologist leave for a job in > Perth, Australia, his name is Dr. Ryan O'Hanley and giardia is his > thing. He was using an Ab for IFATs. > > He bought his Ab's from Waterbourne Inc. in New Orleans. He > ordered "Giardia/A/Glow" Cat # A300. Sounds like a FITC > conjugated Ab, but it's a start. Have a nice weekend. > Greg > > From: "Etheridge, Sandra AGF:EX" > > To: " (histonet@lists.utsouthwestern.edu)" > > Date sent: Thu, 21 Oct 2004 14:04:47 -0700 > Subject: [Histonet] Giardia Primary Antibody > > > Hello everyone, > > > > We are looking for a primary antibody for Giardia. Is anyone out there > in > > the veterinary research community aware of anyone who will supply us > with > > some?? I have checked out numerous websites and found one lab who tests > for > > it in Michigan (from the South Dakota State IHC web page), but have not > > heard back from them. > > > > Any info is, as always, appreciated. > > > > Thanks! > > > > Sandra Etheridge > > > > BC Ministry of Agriculture, Food & Fisheries > > Animal Health Center, Histology > > Abbotsford, BC > > Canada > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > ------- End of forwarded message ------- > > > ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ > Greg Dobbin > Pathology Lab > Atlantic Veterinary College, U.P.E.I. > 550 University Ave. > Charlottetown, P.E.I. > Canada, C1A 4P3 > Phone: (902)566-0744 > Fax: (902)566-0851 > ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ > Happiness is a journey, not a destination. > > > > ------------------------------ > > Message: 2 > Date: Fri, 22 Oct 2004 08:46:48 -0400 > From: mark.lewis@thermo.com > Subject: Re: [Histonet] Microwave retrieval > To: Patti Loykasek > Cc: histonet-bounces@lists.utsouthwestern.edu, histonet > > Message-ID: > > > > Content-Type: text/plain; charset=iso-8859-1 > > > Patty, > > I think you hit the nail on the head when you said "I believe consistency > in what you do is the key to any antigen > retrieval." I believe it is important for all aspects of the Histo lab. It > makes trouble-shooting problems much easier if the work is all done in a > consistent manner. > > Have a nice day > > > Best regards, > > Mark > > Mark Lewis > Product Specialist > Anatomical Pathology > Clinical Diagnostics > Thermo Electron Corporation > (412) 747-4013 > (412) 788-1097 > E-mail: mark.lewis@thermo.com > > > > > > Patti Loykasek > > To: > histonet > > Sent by: cc: > > histonet-bounces@lists.utsouth Subject: Re: > [Histonet] Microwave retrieval > western.edu > > > > > > 10/21/2004 12:09 PM > > > > > > > > > > Ed, > I've done antigen retrieval many ways, and there are pros & cons to > each. I believe consistency in what you do is the key to any antigen > retrieval. I do not recommend microwaving with open containers - it is far > too easy for there to be hot spots, loss of fluid, etc.... That being > said, > I do think it is possible to microwave retrieve in a closed container such > as a microwave pressure cooker. Some of the keys to this technique are: > use > the same volume of liquid, the same # of slides (using blank slides if > necessary) each & every time, monitoring with your microwave the time it > takes to get up to pressure & how long you want your slides to remain > under > pressure, careful cooling down of slides. I1d be happy to provide the > particulars of how we do it, if anyone is interested. In a microwave > pressure cooker the fluid becomes 3super heated2 - much hotter than in a > steamer & you have the added pressure. We have found it improves staining > with many difficult targets & nuclear antigens. > I would note that I work at a reference lab, and our tissue is fixed & > processed many different ways. > If you can control the fixation & processing, and optimize it, I think a > steamer works great & we use it for many antibodies. Again, consistency is > the key. We preheat solutions before adding slides, only adding slides > when > the solution has reached 95 degrees C. To monitor the temperature we have > thermometers with slender, metal probes that fit thru the holes in the > steamer ( alternately, I1ve had my husband carefully enlarge the steamer > lid > holes to accommodate a standard thermometer). After adding slides, we > start > our retrieval time when the solution is back up to 95. We do the same > retrieval time & cool down time each time. > Well, I hope this helps you out. Feel free to contact me for more info. > Good > luck with your retrieval & staining. > > > Patti Loykasek > PhenoPath Laboratories > Seattle, WA > > > > > > Can anyone give me the pro's and con's of steamer/pressure (Decloaker) > vs. > > microwave retrieval ? > > Our Doc wants us to change from the Decloaker to microwave. Is a > household > > microwave adequate or do we need a lab grade unit? > > Does the microwave save time and improve turn around time? > > > > Thanks in advance > > Ed Harris HT ASCP MT ABB > > > > > > > > This E-mail message (including attachments, if any) is intended for the > use of > > the individual or entity to which it is addressed and may contain > information > > that is privileged, proprietary, confidential and exempt from > disclosure. > If > > you are not the intended recipient, you are notified that any > dissemination, > > distribution or copying of this communication is strictly prohibited. If > you > > have received this communication in error, please notify the sender and > erase > > this E-mail message immediately. > > > > _______________________________________________ > > 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 > > > > > > > > > ------------------------------ > > Message: 3 > Date: Fri, 22 Oct 2004 08:42:01 -0500 > From: "Goeden, Catherine" > Subject: [Histonet] CAP question > To: "'histonet@pathology.swmed.edu'" > Message-ID: <597FCC5997E8504EAA6814E6C9ED52298FAD6A@VHASUXEXC1> > Content-Type: text/plain; charset="iso-8859-1" > > > We are getting ready for a CAP inspection and I have a question about how > other techs are handling a question regarding waterbaths: > > ANP.23350 Are they clean and well-maintained, and is there a policy for > preventing cross-contamination of paraffin sections in the bath? > > I have a procedure for cutting the sections and cleaning water baths but I > guess I am not specifically using the word cross-contamination. Would > appreciate how other labs are "wording" this procedure. > > Thanks in advance!! > > > > ------------------------------ > > Message: 4 > Date: Fri, 22 Oct 2004 10:48:40 -0400 (GMT-04:00) > From: amosbrooks@earthlink.net > Subject: RE: [Histonet] Microwave retrieval > To: "SMITH,REBEKAH FELICIA" , > histonet@lists.utsouthwestern.edu > Message-ID: > <10909363.1098456520591.JavaMail.root@skeeter.psp.pas.earthlink.net> > Content-Type: text/plain; charset=us-ascii > > Hi, > Just to point out a steamer is cheaper than a microwave. So for a lab > on a budget the steamer is even better. You should give it a whirl some > time. You may find some improved consistancy. Of course if you are content > with what you have I stand by something I always say: All labs are > different, What works fine in one lab may cause problems in another. If it > aint broke don't fix it. > Amos > > > -----Original Message----- > From: "SMITH,REBEKAH FELICIA" > Sent: Oct 21, 2004 10:29 PM > To: Luis Chiriboga , > "Weems, Joyce" , > marjorie lehman , > Amos Brooks , > histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Microwave retrieval > > Being myself a poor research tech, I also use the household > microwave method. I stick my slides in coplin jars filled with > Tris buffer with the tops off then stick them in another container > and microwave for 45sec twice, changing the water in between. It's > been working for me. > SMITH,REBEKAH FELICIA > "You are a child of the universe, no less than the trees and the > stars > You have a right to be here and whether or not it is clear to you, > no doubt the universe is unfolding as it should. Therefore be at > peace with G-d, whatever you conceive Him to be. And whatever your > labors and aspirations,in the noisy confusion of life, keep peace > in your soul.-Max Ehrmann,"Desiderata" > > > > > ------------------------------ > > Message: 5 > Date: Fri, 22 Oct 2004 09:28:14 -0700 (PDT) > From: Patrice McCall > Subject: [Histonet] reference for plastics > To: histonet@lists.utsouthwestern.edu > Message-ID: <20041022162814.84913.qmail@web54504.mail.yahoo.com> > Content-Type: text/plain; charset=us-ascii > > Does anyone know a good reference book or other > material to use for learning to do plastics in histology? > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - Helps protect you from nasty viruses. > http://promotions.yahoo.com/new_mail > > > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 11, Issue 30 > **************************************** > > From RCazares <@t> schosp.org Fri Oct 22 14:39:17 2004 From: RCazares <@t> schosp.org (Cazares, Ruth) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Billing of Immunos Message-ID: <913FAC2B773C19488E26AE6572180FA53AF68C@exch01.schosp.org> Hello Histonetters, I have a question regarding Immuno billing. If a case has parts 1 & 2, and immunos ( 2 antibodies ) are done on 4 blocks of part 1, how many immuno charges can be billed? There seems to be some disagreement on this. I have always known it to be one charge per specimen per antibody. What are the exceptions, if any ? Thanks, Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. From nancy.troiano <@t> yale.edu Fri Oct 22 14:59:40 2004 From: nancy.troiano <@t> yale.edu (Nancy W. Troiano) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] reference for plastics Message-ID: <5.2.1.1.2.20041022155510.00b38de8@email.med.yale.edu> I don't know what type of plastic you are referring to but if you are interested in methylmethacrylate for undecalcified bone our recent article in JOH, June 2004 (Kacena et al) describes two different methylmethacrylate embedding methods for bone. Also, there is a chapter in a book entitled "Bone Histomorphometry, Techniques and Interpretation, Recker, RR editor, CRC press, 1983 , pp 13-35 that describes another method. Nancy Troiano Yale Core Center for Musculoskeletal Disorders Yale University P.O. Box 208071 New Haven, CT (203)785-5136 From JWEEMS <@t> sjha.org Fri Oct 22 15:05:26 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Billing of Immunos Message-ID: <83AACDB0810528418AA106F9AE9B7F7E27854E@sjhaexc02.sjha.org> You are correct, as I understand it. No exceptions that I am aware of. Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Cazares, Ruth Sent: Friday, October 22, 2004 3:39 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Billing of Immunos Hello Histonetters, I have a question regarding Immuno billing. If a case has parts 1 & 2, and immunos ( 2 antibodies ) are done on 4 blocks of part 1, how many immuno charges can be billed? There seems to be some disagreement on this. I have always known it to be one charge per specimen per antibody. What are the exceptions, if any ? Thanks, Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From JNocito <@t> Pathreflab.com Fri Oct 22 15:17:29 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Billing of Immunos In-Reply-To: <913FAC2B773C19488E26AE6572180FA53AF68C@exch01.schosp.org> Message-ID: Ruth, I just had an in-service on this this morning. You may charge one immuno for each specimen. Example: if I have a case with a cervical bx designated as 12 o'clock and 6 o'clock and the Dr. orders 2 immunos on each block, I charge for 4 immunos. Example 2: If I have a uterus that has 10 blocks and the Dr. asks for 2 immunos on block 1( say cervix) and 2 on block 7 (say endometrium) I can only charge for 2 immunos because there is only 1 specimen despite have multiple areas submitted. If you have any more questions, or if I can explain it better, please don't hesitate to call me 210-231-8058. Hope this clears things up a bit. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Cazares, Ruth Sent: Friday, October 22, 2004 2:39 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Billing of Immunos Hello Histonetters, I have a question regarding Immuno billing. If a case has parts 1 & 2, and immunos ( 2 antibodies ) are done on 4 blocks of part 1, how many immuno charges can be billed? There seems to be some disagreement on this. I have always known it to be one charge per specimen per antibody. What are the exceptions, if any ? Thanks, Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Fri Oct 22 16:35:55 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] RE: detection of HRP in FFPE sections- ?antibody In-Reply-To: <370ba936bab4.36bab4370ba9@amc.uva.nl> Message-ID: <003b01c4b87f$1dc9ac50$83020a0a@IHCTech> I have done DAB on ffpe tissue's to detect endogenous peroxidase (myleoperoxidase) with success to identify neutrophils, just don't use h202 to block for endogenous peroxidase. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of C.M. van der Loos Sent: Thursday, October 21, 2004 12:18 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: detection of HRP in FFPE sections- ?antibody Dear Shaumik, I guess that HRP was used as tracer in some kind of labeling experiment in an whole organ, animal or something, followed by fixation and embedding. As you know endogenous peroxidase activity in erythrocytes and neutrophils perfectly survives the whole embedding circus quite well (unfortunately to many of us). So would try this with your labeling HRP as well. There was a Dako antibody (rabbit) available long time ago. Also at www.biodesign.com. Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Shaumik Adhya Date Thu, 21 Oct 2004 16:42:57 +0100 To Histonet@lists.utsouthwestern.edu Subject [Histonet] detection of HRP in FFPE sections- ?antibody Hi histonetters, I was wondering if any of you out there have experience of detection of HRP in formalin fixed, paraffin embedded tissue. I assume that the process of fixation and tissue processing will destroy the enzymatic activity of HRP, rendering the usual histochemical reagents useless. Are there antibodies out there that anyone's used? Thanks a lot for your help, Shaumik Adhya _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bill501 <@t> mindspring.com Fri Oct 22 18:43:57 2004 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Billing of Immunos In-Reply-To: References: Message-ID: At 3:17 PM -0500 10/22/04, Joe Nocito wrote: >Example 2: If I have a uterus that has 10 blocks and the Dr. asks for 2 >immunos on block 1( say cervix) and 2 on block 7 (say endometrium) Even if you accept the per specimen rational (I don't, I would charge per immuno stainEd slide I have to look at), the Uterus is multiple specimens taken out together - Cervis, endometrium, myometrium are all different organs. It can even be broken down more rigorously (endo- vs exocervix, eg). Of course the bean counters might disagree, but... screw 'em. -- ______________ Bill Blank, MD Heartland Lab From jkiernan <@t> uwo.ca Sat Oct 23 00:19:05 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] RE: detection of HRP in FFPE sections- ?antibody References: <003b01c4b87f$1dc9ac50$83020a0a@IHCTech> Message-ID: <4179E9C9.547C5C4C@uwo.ca> You are quite right. The peroxidase activity of HRP does not survive dehydration and paraffin embedding, but it does survive brief formaldehyde fixation. If you obtain positive "peroxidase" staining in paraffin sections it is probably due to the catalytic action of certain metalloproteins including haemoglobin in erythrocytes and "myeloperoxidase" in leukocytes. Some neurons in the CNS have "endogenous peroxidase" activity that shows up with DAB-H2O2. In neurobiology HRP and HRP-labelled proteins have been used as in vivo tracers for more than 30 years. Endogenous peroxidase-like activity has always been a curse because there is no way to block it without also inhibiting the HRP. The optimal methods for preparing sections were worked out in the 1970s, especially by MM Mesulam and colleagues. Their paper, "Sensitivity in horseradish peroxidase neurohistochemistry: a comparative and quantitative study of nine methods" is a must-read for anyone who needs to detect HRP that has been introduced into a living animal. The paper is in J. Histochem. Cytochem. 27:763-773 (1979). Briefly, you thoroughly perfuse the animal with a neutral buffered formaldehyde solution, put it in a poly bag in the fridge for a few hours, take out the brain and put it in a strong sucrose solution for cryoprotection (24-48h, 4C), then freeze and cut frozen sections. Don't use this summary as a set of instructions. Read Mesulam's paper and some of the earlier ones cited in it. For many neuro purposes thick (50-200 um) sections are needed to see infrequently occurring labelled cells or axons. If there is a high density of HRP labelling, you may need to cut cryostat sections (5-20 um), or cut unfrozen wet sections (100-200 um) with a vibrating micotome, do the DAB-H2O2 peroxidase histochemistry, osmicate, then cut out small areas of interest, and then process these into a resin for ultrathin sectioning and electron microscopy. There is a huge body of literature in this field. It's mostly in journals, and you'll need the full text, not just the PubMed abstract. If you are a researcher you will have access to a library. Use it. If you have already made paraffin sections of your HRP-labelled tissue your only hope may lie with immunohistochemistry directed specifically at the horseradish plant's peroxidase protein. There's probably something to be found if you look in enough catalogues of antibodies and antisera, but it may need many trials to get valid results on your material. It may well be less expensive to repeat the tracing experiments and prepare the tissues optimally for HRP activity. This would also increase the chances of eventually getting the results published in a good journal. -- John A. Kiernan Department of Anatomy & Cell Biology The University of Western Ontario London, Canada. http://publish.uwo.ca/~jkiernan/ ___________________________________________ Shaumik Adhya wrote: > Date Thu, 21 Oct 2004 16:42:57 +0100 > To Histonet@lists.utsouthwestern.edu > Subject [Histonet] detection of HRP in FFPE sections- ?antibody > Hi histonetters, > > I was wondering if any of you out there have experience of detection of > HRP in formalin fixed, paraffin embedded tissue. I assume that the > process of fixation and tissue processing will destroy the enzymatic > activity of HRP, rendering the usual histochemical reagents useless. > Are there antibodies out there that anyone's used? Thanks a lot for > your help, > > Shaumik Adhya > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------------------------- From vanann702 <@t> skmc.gov.ae Sat Oct 23 06:25:57 2004 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] true paperless system Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E02F6D3B8@SKMCEMAIL.skmc.gov.ae> Hi Histonetters Am getting flak from fellow 'clinical techs' who insist that a true paperless system is what I need. Does such a thing really exist in a Histo/Cyto lab. I value your comments Annie in Arabia Anne S van Binsbergen Section Chief, AP Laboratory Shaikh Khalifa Medical Centre PO BOX 51900 Abu Dhabi UAE +971 02 6102695 +971 50 3162804 From g_shaferjp <@t> ybb.ne.jp Sat Oct 23 09:00:46 2004 From: g_shaferjp <@t> ybb.ne.jp (Jen Wyffels & Greg Shafer) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] shark egg carbohydrate jelly staining Message-ID: <003101c4b908$b237e120$2fb28eda@user> I would like to stain the carbohydrate jelly in a shark egg. The egg is similar to a chicken egg, the yolk is surrounded by a protective matrix but in sharks it is carbohydrate rather than albumen. I will remove the yolk leaving the egg jelly within the egg shell. This jelly has 3 distinct and separate compositions, a water like layer (1), a 'snot' like layer (2) and finally a very dense solid (3). I would like to immerse this egg in a carbohydrate stain that would show the layers and their distribution withing the egg case. These layers have been shown (after complete hydrolysis) to differ in the concentration of sugar moeties present with the dense layer containg the highest concentrations. Has anyone tried something similar and willing to share their results? I can experiment a little but egg availability is limited so any advice is welcome. Thanks, Jen From carl.hobbs <@t> kcl.ac.uk Sat Oct 23 13:57:09 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] re microwave.... Message-ID: "If it aint broke don't fix it." Always room for improvement tho........ Lets keep testing! Not just accepting ;-))) --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.778 / Virus Database: 525 - Release Date: 15/10/2004 From jkiernan <@t> uwo.ca Sat Oct 23 23:11:57 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] shark egg carbohydrate jelly staining References: <003101c4b908$b237e120$2fb28eda@user> Message-ID: <417B2B8D.2B57C8F3@uwo.ca> If the biochemistry of shark egg jelly is known (as you indicate), it will probably be possible to localize different carbohydrate components in different colours. Are the carbohydrates bound to protein, as glycoproteins or proteoglycans? If so, they will be fixable with formaldehyde and you will be able to do the staining the usual way - on sections rather than whole specimens. Even if the jelly is protein-free it must consist of very large polysacharide molecules at least in the mucous and solid layers, and can probably insolubilized by a suitable fixative. The choice of staining methods for carbohydrates is quite large, and it's possible to collect significant chemical information. These techniques have all been developed for sections (usually paraffin). Some of the simpler techniques can be applied to very small (0.5 mm) whole objects that are later embedded and sectioned for electron microscopy. How big is a shark's egg? That affects the technology! If you can provide more biochemical information about shark egg jelly, I (and others reading Histonet messages) will be able to advise you about preparative techniques and staining methods. John A. Kiernan Dept of Anatomy & Cell Biology University of Western Ontario London, Canada N6A 5C1 _____________________________________________ Jen Wyffels & Greg Shafer wrote: > > I would like to stain the carbohydrate jelly in a shark egg. The egg is > similar to a chicken egg, the yolk is surrounded by a protective matrix but > in sharks it is carbohydrate rather than albumen. I will remove the yolk > leaving the egg jelly within the egg shell. This jelly has 3 distinct and > separate compositions, a water like layer (1), a 'snot' like layer (2) and > finally a very dense solid (3). I would like to immerse this egg in a > carbohydrate stain that would show the layers and their distribution withing > the egg case. These layers have been shown (after complete hydrolysis) to > differ in the concentration of sugar moeties present with the dense layer > containg the highest concentrations. Has anyone tried something similar and > willing to share their results? I can experiment a little but egg > availability is limited so any advice is welcome. Thanks, Jen > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From g_shaferjp <@t> ybb.ne.jp Sun Oct 24 02:26:05 2004 From: g_shaferjp <@t> ybb.ne.jp (Jen Wyffels & Greg Shafer) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] shark egg carbohydrate jelly staining References: <003101c4b908$b237e120$2fb28eda@user> <417B2B8D.2B57C8F3@uwo.ca> Message-ID: <003501c4b99a$b9b70ce0$2fb28eda@user> Dr. Kiernan and other histonet readers, More information about the egg case jelly in sharks: There has been only one paper that investigated the composition of the 3 different egg jellys in elasmobranch eggs. In that report there was no protein associated with any jelly layer. The different layers were analyzed after hydrolysis for neutral and amino sugars by HPLC and found to contain different proportions and increasing concentrations of the same 4 sugars, glucosamine, galactose, galactosamine and fucose. Note, acidic sugars if present were not measured. The liquid egg jelly (1) was analyzed by gel electrophoresis and had components ranging in size from 31 to 200 kDa as stained by alcian blue, and again no protein was found in the same samples (comasssie blue stain). Size does matter....these eggs are at the smallest 70mm by 20 mm, 15mm thick and the larger eggs are 150mm by 70mm, 35mm thick, both measurements include the egg shell (0.5mm thick). I am afraid I cannot embed and section either egg. I was thinking more of immersing the entire egg after removing the top half of the shell to expose the contents within. I was hoping that the jelly layers would stain differently (either a single stain or multiple) and yield a gradient of color(s) from the center liquid jelly (1) through the next mucus like jelly (2) to the edges of the egg case where the dense jelly (3) is concentrated. I can easily use a fixative on the entire egg, but complete penetration through the jelly may not be possible without disrupting its normal distribution. The dense jelly 3 will not separate from the egg case but the mucus jelly (2) and the liquid jelly (1) are readily lost if the egg is tipped over. I realize the liquid jelly (1) will be lost in any staining technique so I will remove it and attempt to stain it with the same method as the egg but in a test tube. I have already prepared some eggs with embryos in 10% formalin and I can add that the jelly (2) turns from a clear snot to an opaque white color after fixation. The dense jelly (3) is opaque and sometimes has a slight yellow cast before fixation. I hope this information helps, I appreciate the time you have taken to consider this staining adventure. Jen ----- Original Message ----- From: "John Kiernan" To: "Jen Wyffels & Greg Shafer" Cc: Sent: Sunday, October 24, 2004 1:11 PM Subject: Re: [Histonet] shark egg carbohydrate jelly staining > If the biochemistry of shark egg jelly is known > (as you indicate), it will probably be possible to > localize different carbohydrate components in > different colours. Are the carbohydrates bound to > protein, as glycoproteins or proteoglycans? If so, > they will be fixable with formaldehyde and you will > be able to do the staining the usual way - on sections > rather than whole specimens. Even if the jelly is > protein-free it must consist of very large polysacharide > molecules at least in the mucous and solid layers, and > can probably insolubilized by a suitable fixative. > > The choice of staining methods for carbohydrates is > quite large, and it's possible to collect significant > chemical information. These techniques have all > been developed for sections (usually paraffin). Some > of the simpler techniques can be applied to very small > (0.5 mm) whole objects that are later embedded and > sectioned for electron microscopy. > > How big is a shark's egg? That affects the technology! > If you can provide more biochemical information about > shark egg jelly, I (and others reading Histonet > messages) will be able to advise you about preparative > techniques and staining methods. > > John A. Kiernan > Dept of Anatomy & Cell Biology > University of Western Ontario > London, Canada N6A 5C1 > _____________________________________________ > Jen Wyffels & Greg Shafer wrote: > > > > I would like to stain the carbohydrate jelly in a shark egg. The egg is > > similar to a chicken egg, the yolk is surrounded by a protective matrix but > > in sharks it is carbohydrate rather than albumen. I will remove the yolk > > leaving the egg jelly within the egg shell. This jelly has 3 distinct and > > separate compositions, a water like layer (1), a 'snot' like layer (2) and > > finally a very dense solid (3). I would like to immerse this egg in a > > carbohydrate stain that would show the layers and their distribution withing > > the egg case. These layers have been shown (after complete hydrolysis) to > > differ in the concentration of sugar moeties present with the dense layer > > containg the highest concentrations. Has anyone tried something similar and > > willing to share their results? I can experiment a little but egg > > availability is limited so any advice is welcome. Thanks, Jen > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From contact <@t> excaliburpathology.com Sun Oct 24 13:57:12 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Shark egg Message-ID: <20041024185712.68332.qmail@web50303.mail.yahoo.com> I agree that the size of the specimen will affect the processing schedule and fixation will allow paraffin processing procedure. The amount of water naturally occuring in the egg white may cause some shrinkage as it is removed during processing, but you should still get a fairly decent section. No bone in sharks, all cartilage. The eggs are not covered by a calcified shell such as avian eggs, so are you fixing the whole egg, then disecting? What is the pH of the different layers? I am thinking a combo alcian blue-PAS stain would show the different layers very nicely by tweeking the pH of the alcian blue. Basic layers will be blue, acidic will be hot pink, and neutral will be purple. Counterstain with hematoxylin. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From amosbrooks <@t> earthlink.net Sun Oct 24 17:44:56 2004 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Changes to methodology In-Reply-To: <200410241001.1clLK8MI3NZFpP0@mx-a065b23.pas.sa.earthlink.n et> References: <200410241001.1clLK8MI3NZFpP0@mx-a065b23.pas.sa.earthlink.net> Message-ID: <6.0.0.22.0.20041024182704.01c5a310@pop.earthlink.net> Hi, You know, ya just can't please anyone. I've sent messages to the list in the past preaching the fire and brimstone of certain methods of processing tissue or staining it or some such thing only to be berated by both colleges and vendors for either being narrow minded for not seeing alternative ways of doing things. So, being a good person I adapted my approach and when we do something differently I stated so and explained this as best I could with a correlary that if you want to do it differently that's OK just make sure that you get consistent results. Now people are basically saying that I'm advocating histology anarchy! Now it's "We can't have variation in the labs. We need uniformity and structure!" OK, which is it folks. Should we say something is not right if we have seen problems with it in the past, or should we say people are people and will do things differently? If you say it is wrong you are accused of being Machiavelli incarnate. If you say you do things differently but don't chastise them for not conforming to your way you are accused of being a "flip flopper". I'm confused... Amos From pengbw <@t> sjtu.edu.cn Mon Oct 25 02:01:10 2004 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] need protocol for EM Message-ID: <20041025070110.BD57511101F6@sjtu.edu.cn> Dear all netters, Does anyone can share me a protocol for EM to observe tissue sections? Baowei Peng 1954 Huashan Road Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China From pengbw <@t> sjtu.edu.cn Mon Oct 25 10:05:17 2004 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] need protocol for EM Message-ID: <20041025150517.B3D2E10EC931@sjtu.edu.cn> Thanks for all replied to my post. Indeed I have no idea about using EM to observe tissue sections. What I want to do is using EM to trace a particle which is belived to be taken up by macrophage/DC in lymph node, the tracer have been said can be observed under EM. So I need a protocol in which I can learn from tissue removal through all steps to embedding. I only have a machine can cut sections to 5 micrometer thick. But maybe I can get these material include grids from EM technologist. From tpmorken <@t> labvision.com Mon Oct 25 10:56:54 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] true paperless system Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA257F632@usca0082k08.labvision.apogent.com> Anne, It can, but it requires several "engineering" solutions. 1) A common data storage system for the whole institution. That is so rare right now that you could probably count those systems on one hand - woprldwide. 2) universal access to a computer/terminal. For instance, how is the cutting or stains person going to know what to do with a given block or slide? Paper or a screen representation is needed (a la fast food restaurants - every look at the screens the burder flippers get their instructions from?). This also applies to offices or any place that people discuss cases - they have to be able to access the information from anywhere. Wireless networks and tablet computers and pda's come to mind. 3) scan every bit of paper that comes in with a case so it can be viewed on screen (assuming the rest of the hospital is also paperless (BTW - do the clinicians order everything online?)). 4) Barcode everything - including every slide cut. Most of the current LIS's do not allow identification of individual slides, so not much hope with those. 5) Are paper "backup" records necessary? Only if your computer system is not failproof. Anyone want to bet on that? And even if it is, the records have to kept in a medium that can be read in the future. That means transferring to new and/or updated storage medium every few years. It would be interesting to know how far along your clincial collegues really are in this quest. My guess is, not as far as they like to think. Tim Morken -----Original Message----- From: Anne Van Binsbergen [mailto:vanann702@skmc.gov.ae] Sent: Saturday, October 23, 2004 4:26 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] true paperless system Hi Histonetters Am getting flak from fellow 'clinical techs' who insist that a true paperless system is what I need. Does such a thing really exist in a Histo/Cyto lab. I value your comments Annie in Arabia Anne S van Binsbergen Section Chief, AP Laboratory Shaikh Khalifa Medical Centre PO BOX 51900 Abu Dhabi UAE +971 02 6102695 +971 50 3162804 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MAUGER <@t> email.chop.edu Mon Oct 25 11:06:26 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] eosin in 95% Message-ID: Dear list, Those of you who use eosin in your 95% etoh on the processor-what exactly do you use? The ready to stain eosin, or more concentrated? I remember someone writing that they put it in the first 95,correct? We need to stain our tiny gi bxs to better see them when embedding and cutting. Any tips are appreciated. Jo >>> "Melanie Black" 10/21/04 02:28AM >>> >I have also had rod-like crystals in my Alk phos/NBT system. I have >shown them to the company rep, who has no idea what they might be. >With so much written about it here...I think DAKO should sit up and >take a look. Melanie Black > >Thanks all for the cat scratch advice. I am trying it with HRP- AEC. >Will report result. >Jo > > >>>> "C.M. van der Loos" 10/17/04 02:20PM >>> >Dear Joanne, > >Usually, AP staining doesn't show up in a crystal-like precipitate, >but I have seen it a couple of times. Never found out what was the >cause of it. Be sure that after your IHC procedure and before the AP >visualization, the sections are washed at with a Tris-HCl pH8.5 >buffer. Phosphate ions from PBS will inhibit your AP reaction. You >also may mix the Permanent Red components first and than press >immediately through a syringe-filter to remove all bigger particles >in the AP solution. >Have you viewed your slides under a fluorescence microscope? A small >amount of Permanent Red reaction product also gives a bright red >fluorescence signal when using the rhodamine filter pack (green >light). >Good suggestion by Richard to use a tonsil as a negative control. >But, how about running the good old control blanc slide without any >primary antibody? >Hope this helps. > >Chris van der Loos, PhD >Dept. of Pathology >Academical Medical Center >Amsterdam - The Netherlands > >----- Original Message ----- >>From "Joanne Mauger" >Date Fri, 15 Oct 2004 15:45:05 -0400 >To histonet@lists.utsouthwestern.edu, jkiernan@uwo.ca >Subject Re: [Histonet] IHC withquantum dots >Hi group, >Can anyone help me with suggestions-I am working up the monoclonal >Ab (H2A10) to cat scratch disease. Ab from Novus Biologicus. I have >a purchased control from Newcomer Supply. I am using permanent red >chromagen from Dako., with alk phos link also from Dako. I am >getting a nice specific staining of what looks like specs vs tiny >rods or spirochetes, but the control tissue does , by Warthin >Starry, look much more spirochete-ish. Can I trust this result? I >have also stained several suspected + cases and see the same dust >like + staining,but I am afraid it is teeny bits of precipitate. >Does this ring a bell? >Please help. >Thanks, >Jo > >----- Original Message ----- >>From "Richard Cartun" >Date Fri, 15 Oct 2004 16:45:57 -0400 >To ,, > >Subject Re: [Histonet] IHC withquantum dots >Have you tried staining a section of tonsil to see if you get the >same pattern of staining. I have found "precipitation" to be a >problem with alkaline phosphatase (PA) detection, although I am by >no means an expert when it comes to AP IHC. I use HRP/AEC for most >of my infectious disease studies including Bartonella henselae. >Don't forget that you may not see the normal morphology of the >bacterium with IHC. You are labeling epitopes on the surface of the >organism; you are not staining the entire cell wall. I have found >B. henselae to be one of the most difficult organisms to label (and >interpret) with IHC. Good luck! > >Richard > > >Richard W. Cartun, Ph.D. >Director, Immunopathology & Histology >Assistant Director, Anatomic Pathology >Hartford Hospital >80 Seymour Street >Hartford, CT 06102 > > > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Cardiovascular Research Unit Div. of Cardiothoracic Surgery Chris Barnard Building University of Cape Town Anzio Road Observatory 7925 Republic of South Africa Tel +27 21 406-6589 Cel +27 82 469-3352 Fax +27 21 448-5935 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Nancy.Temple <@t> ssfhs.org Mon Oct 25 11:36:39 2004 From: Nancy.Temple <@t> ssfhs.org (Temple Nancy) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] eosin in 95% Message-ID: <4862D47C7409844E9E52532B25609062010153D3@ontexind01.ssfhs.org> We put about 20-30 ml. of our regular eosin (from H&E stain) in the final alcohol(100%) on both our processors. This helps stain not only our GI bx. but also prostate needle bx. or any other small possibly hard to see bx. Nancy SFHC -----Original Message----- From: Joanne Mauger [mailto:MAUGER@email.chop.edu] Sent: Monday, October 25, 2004 11:06 AM To: ctsblack@capeheart.uct.ac.za; histonet@pathology.swmed.edu Subject: [Histonet] eosin in 95% Dear list, Those of you who use eosin in your 95% etoh on the processor-what exactly do you use? The ready to stain eosin, or more concentrated? I remember someone writing that they put it in the first 95,correct? We need to stain our tiny gi bxs to better see them when embedding and cutting. Any tips are appreciated. Jo >>> "Melanie Black" 10/21/04 02:28AM >>> >I have also had rod-like crystals in my Alk phos/NBT system. I have >shown them to the company rep, who has no idea what they might be. >With so much written about it here...I think DAKO should sit up and >take a look. Melanie Black > >Thanks all for the cat scratch advice. I am trying it with HRP- AEC. >Will report result. >Jo > > >>>> "C.M. van der Loos" 10/17/04 02:20PM >>> >Dear Joanne, > >Usually, AP staining doesn't show up in a crystal-like precipitate, >but I have seen it a couple of times. Never found out what was the >cause of it. Be sure that after your IHC procedure and before the AP >visualization, the sections are washed at with a Tris-HCl pH8.5 >buffer. Phosphate ions from PBS will inhibit your AP reaction. You >also may mix the Permanent Red components first and than press >immediately through a syringe-filter to remove all bigger particles >in the AP solution. >Have you viewed your slides under a fluorescence microscope? A small >amount of Permanent Red reaction product also gives a bright red >fluorescence signal when using the rhodamine filter pack (green >light). >Good suggestion by Richard to use a tonsil as a negative control. >But, how about running the good old control blanc slide without any >primary antibody? >Hope this helps. > >Chris van der Loos, PhD >Dept. of Pathology >Academical Medical Center >Amsterdam - The Netherlands > >----- Original Message ----- >>From "Joanne Mauger" >Date Fri, 15 Oct 2004 15:45:05 -0400 >To histonet@lists.utsouthwestern.edu, jkiernan@uwo.ca >Subject Re: [Histonet] IHC withquantum dots >Hi group, >Can anyone help me with suggestions-I am working up the monoclonal >Ab (H2A10) to cat scratch disease. Ab from Novus Biologicus. I have >a purchased control from Newcomer Supply. I am using permanent red >chromagen from Dako., with alk phos link also from Dako. I am >getting a nice specific staining of what looks like specs vs tiny >rods or spirochetes, but the control tissue does , by Warthin >Starry, look much more spirochete-ish. Can I trust this result? I >have also stained several suspected + cases and see the same dust >like + staining,but I am afraid it is teeny bits of precipitate. >Does this ring a bell? >Please help. >Thanks, >Jo > >----- Original Message ----- >>From "Richard Cartun" >Date Fri, 15 Oct 2004 16:45:57 -0400 >To ,, > >Subject Re: [Histonet] IHC withquantum dots >Have you tried staining a section of tonsil to see if you get the >same pattern of staining. I have found "precipitation" to be a >problem with alkaline phosphatase (PA) detection, although I am by >no means an expert when it comes to AP IHC. I use HRP/AEC for most >of my infectious disease studies including Bartonella henselae. >Don't forget that you may not see the normal morphology of the >bacterium with IHC. You are labeling epitopes on the surface of the >organism; you are not staining the entire cell wall. I have found >B. henselae to be one of the most difficult organisms to label (and >interpret) with IHC. Good luck! > >Richard > > >Richard W. Cartun, Ph.D. >Director, Immunopathology & Histology >Assistant Director, Anatomic Pathology >Hartford Hospital >80 Seymour Street >Hartford, CT 06102 > > > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Cardiovascular Research Unit Div. of Cardiothoracic Surgery Chris Barnard Building University of Cape Town Anzio Road Observatory 7925 Republic of South Africa Tel +27 21 406-6589 Cel +27 82 469-3352 Fax +27 21 448-5935 _______________________________________________ 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 ____________________________________________________________________________ __________________________________ The information contained in this email and any accompanying documents is intended for the sole use of the recipient to whom it is addressed, and may contain information that is privileged, confidential, and prohibited from disclosure under applicable law. If you are not the intended recipient, or authorized to receive this on behalf of the recipient, you are hereby notified that any review, use, disclosure, copying, or distribution is prohibited. If you are not the intended recipient(s), please contact the sender by e-mail and destroy all copies of the original message. Thank you. From BlazekL <@t> childrensdayton.org Mon Oct 25 11:45:07 2004 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] eosin in 95% Message-ID: We put the eosin in the last 100%. We use about 10 - 20 ml from the same eosin we use on our staining set up. I don't think it really makes a lot of difference if you put it in the 95% or the 100% and the volume of eosin is pretty much a personal preference. It doesn't take very much to get the tint in the biopsies. Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center 1 Children's Plaza Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org >>> "Joanne Mauger" 10/25/2004 12:06:26 PM >>> Dear list, Those of you who use eosin in your 95% etoh on the processor-what exactly do you use? The ready to stain eosin, or more concentrated? I remember someone writing that they put it in the first 95,correct? We need to stain our tiny gi bxs to better see them when embedding and cutting. Any tips are appreciated. Jo From gu.lang <@t> gmx.at Mon Oct 25 11:55:38 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] eosin in 95% References: Message-ID: <003d01c4bab3$7473e080$eeeea8c0@server> We have another aproach to this. We give one or two drops of 2% Eosin in the biopsy-container in the formalin, if the bx is very tiny. So only the biopsy will be stained and not the whole number of blocks. On white paper the tissue is then well seen. Gudrun ----- Original Message ----- From: "Joanne Mauger" To: ; Sent: Monday, October 25, 2004 6:06 PM Subject: [Histonet] eosin in 95% Dear list, Those of you who use eosin in your 95% etoh on the processor-what exactly do you use? The ready to stain eosin, or more concentrated? I remember someone writing that they put it in the first 95,correct? We need to stain our tiny gi bxs to better see them when embedding and cutting. Any tips are appreciated. Jo >>> "Melanie Black" 10/21/04 02:28AM >>> >I have also had rod-like crystals in my Alk phos/NBT system. I have >shown them to the company rep, who has no idea what they might be. >With so much written about it here...I think DAKO should sit up and >take a look. Melanie Black > >Thanks all for the cat scratch advice. I am trying it with HRP- AEC. >Will report result. >Jo > > >>>> "C.M. van der Loos" 10/17/04 02:20PM >>> >Dear Joanne, > >Usually, AP staining doesn't show up in a crystal-like precipitate, >but I have seen it a couple of times. Never found out what was the >cause of it. Be sure that after your IHC procedure and before the AP >visualization, the sections are washed at with a Tris-HCl pH8.5 >buffer. Phosphate ions from PBS will inhibit your AP reaction. You >also may mix the Permanent Red components first and than press >immediately through a syringe-filter to remove all bigger particles >in the AP solution. >Have you viewed your slides under a fluorescence microscope? A small >amount of Permanent Red reaction product also gives a bright red >fluorescence signal when using the rhodamine filter pack (green >light). >Good suggestion by Richard to use a tonsil as a negative control. >But, how about running the good old control blanc slide without any >primary antibody? >Hope this helps. > >Chris van der Loos, PhD >Dept. of Pathology >Academical Medical Center >Amsterdam - The Netherlands > >----- Original Message ----- >>From "Joanne Mauger" >Date Fri, 15 Oct 2004 15:45:05 -0400 >To histonet@lists.utsouthwestern.edu, jkiernan@uwo.ca >Subject Re: [Histonet] IHC withquantum dots >Hi group, >Can anyone help me with suggestions-I am working up the monoclonal >Ab (H2A10) to cat scratch disease. Ab from Novus Biologicus. I have >a purchased control from Newcomer Supply. I am using permanent red >chromagen from Dako., with alk phos link also from Dako. I am >getting a nice specific staining of what looks like specs vs tiny >rods or spirochetes, but the control tissue does , by Warthin >Starry, look much more spirochete-ish. Can I trust this result? I >have also stained several suspected + cases and see the same dust >like + staining,but I am afraid it is teeny bits of precipitate. >Does this ring a bell? >Please help. >Thanks, >Jo > >----- Original Message ----- >>From "Richard Cartun" >Date Fri, 15 Oct 2004 16:45:57 -0400 >To ,, > >Subject Re: [Histonet] IHC withquantum dots >Have you tried staining a section of tonsil to see if you get the >same pattern of staining. I have found "precipitation" to be a >problem with alkaline phosphatase (PA) detection, although I am by >no means an expert when it comes to AP IHC. I use HRP/AEC for most >of my infectious disease studies including Bartonella henselae. >Don't forget that you may not see the normal morphology of the >bacterium with IHC. You are labeling epitopes on the surface of the >organism; you are not staining the entire cell wall. I have found >B. henselae to be one of the most difficult organisms to label (and >interpret) with IHC. Good luck! > >Richard > > >Richard W. Cartun, Ph.D. >Director, Immunopathology & Histology >Assistant Director, Anatomic Pathology >Hartford Hospital >80 Seymour Street >Hartford, CT 06102 > > > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Cardiovascular Research Unit Div. of Cardiothoracic Surgery Chris Barnard Building University of Cape Town Anzio Road Observatory 7925 Republic of South Africa Tel +27 21 406-6589 Cel +27 82 469-3352 Fax +27 21 448-5935 _______________________________________________ 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 From tdobersztyn <@t> chmca.org Mon Oct 25 12:24:50 2004 From: tdobersztyn <@t> chmca.org (tdobersztyn@chmca.org) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] EM protocol Message-ID: I am an EM tech here in Akron, Ohio doing EM on medical kidney, tumor, muscle, &nerve biopsies.. What exactly do you need??? processing? Microtomy? I have info on all EM techniques. Theresa R Dobersztyn HT ASCP Electron Microscopy Laboratory Department of Pathology Akron Children's Hospital 1 Perkins Square Akron, Ohio 44308 From TJJ <@t> Stowers-Institute.org Mon Oct 25 12:34:04 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Changes to methodology Message-ID: Amos, You've just summed up the political climate in the United States perfectly.... Frankly, I believe that methodology should be followed as written. Protocol development is where you can make changes, starting with published protocols and making whatever modifications from there. Controls, controls, and controls and very critical evaluation should then follow. Best of both worlds. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From JNocito <@t> Pathreflab.com Mon Oct 25 14:46:47 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] eosin in 95% In-Reply-To: Message-ID: Joanne, we put about 30 mLs of our regular staining eosin in our first 100% alcohol. We used less, but my embedders thought it was too light still. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joanne Mauger Sent: Monday, October 25, 2004 11:06 AM To: ctsblack@capeheart.uct.ac.za; histonet@pathology.swmed.edu Subject: [Histonet] eosin in 95% Dear list, Those of you who use eosin in your 95% etoh on the processor-what exactly do you use? The ready to stain eosin, or more concentrated? I remember someone writing that they put it in the first 95,correct? We need to stain our tiny gi bxs to better see them when embedding and cutting. Any tips are appreciated. Jo >>> "Melanie Black" 10/21/04 02:28AM >>> >I have also had rod-like crystals in my Alk phos/NBT system. I have >shown them to the company rep, who has no idea what they might be. >With so much written about it here...I think DAKO should sit up and >take a look. Melanie Black > >Thanks all for the cat scratch advice. I am trying it with HRP- AEC. >Will report result. >Jo > > >>>> "C.M. van der Loos" 10/17/04 02:20PM >>> >Dear Joanne, > >Usually, AP staining doesn't show up in a crystal-like precipitate, >but I have seen it a couple of times. Never found out what was the >cause of it. Be sure that after your IHC procedure and before the AP >visualization, the sections are washed at with a Tris-HCl pH8.5 >buffer. Phosphate ions from PBS will inhibit your AP reaction. You >also may mix the Permanent Red components first and than press >immediately through a syringe-filter to remove all bigger particles >in the AP solution. >Have you viewed your slides under a fluorescence microscope? A small >amount of Permanent Red reaction product also gives a bright red >fluorescence signal when using the rhodamine filter pack (green >light). >Good suggestion by Richard to use a tonsil as a negative control. >But, how about running the good old control blanc slide without any >primary antibody? >Hope this helps. > >Chris van der Loos, PhD >Dept. of Pathology >Academical Medical Center >Amsterdam - The Netherlands > >----- Original Message ----- >>From "Joanne Mauger" >Date Fri, 15 Oct 2004 15:45:05 -0400 >To histonet@lists.utsouthwestern.edu, jkiernan@uwo.ca >Subject Re: [Histonet] IHC withquantum dots >Hi group, >Can anyone help me with suggestions-I am working up the monoclonal >Ab (H2A10) to cat scratch disease. Ab from Novus Biologicus. I have >a purchased control from Newcomer Supply. I am using permanent red >chromagen from Dako., with alk phos link also from Dako. I am >getting a nice specific staining of what looks like specs vs tiny >rods or spirochetes, but the control tissue does , by Warthin >Starry, look much more spirochete-ish. Can I trust this result? I >have also stained several suspected + cases and see the same dust >like + staining,but I am afraid it is teeny bits of precipitate. >Does this ring a bell? >Please help. >Thanks, >Jo > >----- Original Message ----- >>From "Richard Cartun" >Date Fri, 15 Oct 2004 16:45:57 -0400 >To ,, > >Subject Re: [Histonet] IHC withquantum dots >Have you tried staining a section of tonsil to see if you get the >same pattern of staining. I have found "precipitation" to be a >problem with alkaline phosphatase (PA) detection, although I am by >no means an expert when it comes to AP IHC. I use HRP/AEC for most >of my infectious disease studies including Bartonella henselae. >Don't forget that you may not see the normal morphology of the >bacterium with IHC. You are labeling epitopes on the surface of the >organism; you are not staining the entire cell wall. I have found >B. henselae to be one of the most difficult organisms to label (and >interpret) with IHC. Good luck! > >Richard > > >Richard W. Cartun, Ph.D. >Director, Immunopathology & Histology >Assistant Director, Anatomic Pathology >Hartford Hospital >80 Seymour Street >Hartford, CT 06102 > > > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Cardiovascular Research Unit Div. of Cardiothoracic Surgery Chris Barnard Building University of Cape Town Anzio Road Observatory 7925 Republic of South Africa Tel +27 21 406-6589 Cel +27 82 469-3352 Fax +27 21 448-5935 _______________________________________________ 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 From swaram <@t> myrealbox.com Mon Oct 25 20:41:36 2004 From: swaram <@t> myrealbox.com (Swaram) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] PNA Telomere Probe In-Reply-To: <20041019014858.5798110CD52D@sjtu.edu.cn> References: <20041019014858.5798110CD52D@sjtu.edu.cn> Message-ID: <417DAB50.6000904@myrealbox.com> Hello Histonetters, I am using Cy3-PNA Telomere Probe from ABI on 8-10 year old paraffin fixed tissues. I also have a Cy5-Centromere probe in the same mix. I deparaffinize, treat with NaSCN 80°C (4min), Pepsin 37°C (8min), air dry and then denature in denaturing solution 74°C, air dry again, keep sections on heating block 37°C and apply 10µl of the Probe mix, cover with a small piece of plastic coverslip, heat to 81°C in a Hybrite and then leave OVT at 37°C in a humidity chamber. Washing is done in 50% CH3-NO, 2X SSC and 2X SSC-Tween-20, mounted with DAPI and Prolong antifade. However, I get good signals of the Cy5-centromere probe, but mostly no signals of the Telomere Probe. Sometimes, in the same tissue I get the Telomere probe signals (faint) but then I also see some non specific signals in the cytoplasm. I have run controls using metaphase spreads and verified that the Cy3-Telomere probe works fine. Please help me overcome the high background and non specific signals in the Cy3-Telomere PNA Probe. Interestingly, the first time, I ran this experiment, I got really good signals on both Cy3-Cy5. But I started getting problems after I changed the lot of the probe. Sincerely, Swaram Univ. California From swaram <@t> myrealbox.com Mon Oct 25 20:44:36 2004 From: swaram <@t> myrealbox.com (Swaram) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] PNA Telomere Probe In-Reply-To: <20041019014858.5798110CD52D@sjtu.edu.cn> References: <20041019014858.5798110CD52D@sjtu.edu.cn> Message-ID: <417DAC04.7090402@myrealbox.com> Hello Histonetters, I am using Cy3-PNA Telomere Probe from ABI on 8-10 year old paraffin fixed tissues. I also have a Cy5-Centromere probe in the same mix. I deparaffinize, treat with NaSCN 80°C (4min), Pepsin 37°C (8min), air dry and then denature in denaturing solution 74°C, air dry again, keep sections on heating block 37°C and apply 10µl of the Probe mix, cover with a small piece of plastic coverslip, heat to 81°C in a Hybrite and then leave OVT at 37°C in a humidity chamber. Washing is done in 50% CH3-NO, 2X SSC and 2X SSC-Tween-20, mounted with DAPI and Prolong antifade. However, I get good signals of the Cy5-centromere probe, but mostly no signals of the Telomere Probe. Sometimes, in the same tissue I get the Telomere probe signals (faint) but then I also see some non specific signals in the cytoplasm. I have run controls using metaphase spreads and verified that the Cy3-Telomere probe works fine. Please help me overcome the high background and non specific signals in the Cy3-Telomere PNA Probe. Interestingly, the first time, I ran this experiment, I got really good signals on both Cy3-Cy5. But I started getting problems after I changed the lot of the probe. Sincerely, Swaram Univ. California From emry <@t> u.washington.edu Mon Oct 25 21:18:12 2004 From: emry <@t> u.washington.edu (P. Emry) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] I'm Sorry! Message-ID: Dear Histoneters, Last week I attempted some humor in asking for some recommendations for staining bone sutures (beyond H&E) and got not one reply. If I offened..please forgive me. I had just gotten my car stolen and was in a state. (Got the car back a bit banged up, but runs). I would appreciate some suggestions. Thank you, Trisha From ClarkatSCSC <@t> aol.com Mon Oct 25 23:26:46 2004 From: ClarkatSCSC <@t> aol.com (ClarkatSCSC@aol.com) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Looking for a Cryostat for Mohs Message-ID: <30.63e551b3.2eaf2c06@aol.com> Hi Everyone, I'm looking for a great cryostat for Mohs. I'm currently using a Tissue Tek thats about 30 yrs old and love it. I have tried a couple new ones on the market but nothing can compare. Does anyone feel that they have the best cryostat out there for Mohs. I would love to hear from you. Thanks. David Clark Portland, Or From jkiernan <@t> uwo.ca Tue Oct 26 00:10:53 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] I'm Sorry! References: Message-ID: <417DDC5D.A40FB703@uwo.ca> Sympathy ++ for the car. Your attempt for compensation by way of humour can't have worked, but cannot have offended anyone. Probably your humorous effort was so subtle and clever that it got passed by as yet another unanswerable query. I hope you get real compensation from your car's insurance company. Don't apologize to histonetters! If you didn't get even one reply it must mean you didn't offend anyone. It may also mean that your message was so smart that none of the histonetters (how many now, Herb? 2000?) picked up on the point you were trying to make. This happens all the time: send a provocative message - nobody takes you up on it. John Kiernan London, Canada. ------------------------------------------------ "P. Emry" wrote: > > Dear Histoneters, > > Last week I attempted some humor in asking for some recommendations for > staining bone sutures (beyond H&E) and got not one reply. > > If I offended..please forgive me. I had just gotten my car stolen and was > in a state. (Got the car back a bit banged up, but runs). > > I would appreciate some suggestions. > > Thank you, > > Trisha > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Mon Oct 25 10:46:18 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] shark egg carbohydrate jelly staining References: <003101c4b908$b237e120$2fb28eda@user> <417B2B8D.2B57C8F3@uwo.ca> <003501c4b99a$b9b70ce0$2fb28eda@user> Message-ID: <417D1FCA.19FF3D9F@uwo.ca> The sugars you name in the first paragraph are ones that go with glycoproteins. Even though acidic sugars were not mentioned in the "only one paper," they must be present in the components separated by electrophoresis, or alcian blue staining would not occur, because this dye stains only sugar-acids. The change to opaque white in formaldehyde indicates that protein is present because formaldehyde does not react chemically with most carbohydrates; it does cross-link and insolubilize proteins. Your eggs are quite large specimens, and it's difficult to imagine a dye penetrating evenly to the centre and forming a meaningful gradient of colour. If you do not have the equipment or expertise to make sections your best bet may be to collaborate with someone who has. Marine biology labs must surely do that kind of work. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Jen Wyffels & Greg Shafer wrote: > > Dr. Kiernan and other histonet readers, > > More information about the egg case jelly in sharks: > > There has been only one paper that investigated the composition of the 3 > different egg jellys in elasmobranch eggs. In that report there was no > protein associated with any jelly layer. The different layers were analyzed > after hydrolysis for neutral and amino sugars by HPLC and found to contain > different proportions and increasing concentrations of the same 4 sugars, > glucosamine, galactose, galactosamine and fucose. Note, acidic sugars if > present were not measured. The liquid egg jelly (1) was analyzed by gel > electrophoresis and had components ranging in size from 31 to 200 kDa as > stained by alcian blue, and again no protein was found in the same samples > (comasssie blue stain). > > Size does matter....these eggs are at the smallest 70mm by 20 mm, 15mm thick > and the larger eggs are 150mm by 70mm, 35mm thick, both measurements include > the egg shell (0.5mm thick). I am afraid I cannot embed and section either > egg. I was thinking more of immersing the entire egg after removing the top > half of the shell to expose the contents within. I was hoping that the > jelly layers would stain differently (either a single stain or multiple) and > yield a gradient of color(s) from the center liquid jelly (1) through the > next mucus like jelly (2) to the edges of the egg case where the dense jelly > (3) is concentrated. I can easily use a fixative on the entire egg, but > complete penetration through the jelly may not be possible without > disrupting its normal distribution. The dense jelly 3 will not separate > from the egg case but the mucus jelly (2) and the liquid jelly (1) are > readily lost if the egg is tipped over. I realize the liquid jelly (1) will > be lost in any staining technique so I will remove it and attempt to stain > it with the same method as the egg but in a test tube. I have already > prepared some eggs with embryos in 10% formalin and I can add that the jelly > (2) turns from a clear snot to an opaque white color after fixation. The > dense jelly (3) is opaque and sometimes has a slight yellow cast before > fixation. > > I hope this information helps, I appreciate the time you have taken to > consider this staining adventure. Jen > > ----- Original Message ----- > From: "John Kiernan" > To: "Jen Wyffels & Greg Shafer" > Cc: > Sent: Sunday, October 24, 2004 1:11 PM > Subject: Re: [Histonet] shark egg carbohydrate jelly staining > > > If the biochemistry of shark egg jelly is known > > (as you indicate), it will probably be possible to > > localize different carbohydrate components in > > different colours. Are the carbohydrates bound to > > protein, as glycoproteins or proteoglycans? If so, > > they will be fixable with formaldehyde and you will > > be able to do the staining the usual way - on sections > > rather than whole specimens. Even if the jelly is > > protein-free it must consist of very large polysacharide > > molecules at least in the mucous and solid layers, and > > can probably insolubilized by a suitable fixative. > > > > The choice of staining methods for carbohydrates is > > quite large, and it's possible to collect significant > > chemical information. These techniques have all > > been developed for sections (usually paraffin). Some > > of the simpler techniques can be applied to very small > > (0.5 mm) whole objects that are later embedded and > > sectioned for electron microscopy. > > > > How big is a shark's egg? That affects the technology! > > If you can provide more biochemical information about > > shark egg jelly, I (and others reading Histonet > > messages) will be able to advise you about preparative > > techniques and staining methods. > > > > John A. Kiernan > > Dept of Anatomy & Cell Biology > > University of Western Ontario > > London, Canada N6A 5C1 > > _____________________________________________ > > Jen Wyffels & Greg Shafer wrote: > > > > > > I would like to stain the carbohydrate jelly in a shark egg. The egg is > > > similar to a chicken egg, the yolk is surrounded by a protective matrix > but > > > in sharks it is carbohydrate rather than albumen. I will remove the > yolk > > > leaving the egg jelly within the egg shell. This jelly has 3 distinct > and > > > separate compositions, a water like layer (1), a 'snot' like layer (2) > and > > > finally a very dense solid (3). I would like to immerse this egg in a > > > carbohydrate stain that would show the layers and their distribution > withing > > > the egg case. These layers have been shown (after complete hydrolysis) > to > > > differ in the concentration of sugar moeties present with the dense > layer > > > containg the highest concentrations. Has anyone tried something similar > and > > > willing to share their results? I can experiment a little but egg > > > availability is limited so any advice is welcome. Thanks, Jen > > > > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From slimwillie <@t> cox.net Tue Oct 26 01:23:23 2004 From: slimwillie <@t> cox.net (Jerry Wilson) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] CAP question References: <597FCC5997E8504EAA6814E6C9ED52298FAD6A@VHASUXEXC1> Message-ID: <006401c4bb24$4c39f520$702e6044@no.cox.net> We word it exactly like the question. Included in the procedure manual and on the daily temp. chart there is a statement that goes something like: "To avoid cross contamination of sections, the water bath is wiped clean with Kim-wipes between each section." Hope this helps Jerry ----- Original Message ----- From: "Goeden, Catherine" To: Sent: Friday, October 22, 2004 8:42 AM Subject: [Histonet] CAP question > > We are getting ready for a CAP inspection and I have a question about how > other techs are handling a question regarding waterbaths: > > ANP.23350 Are they clean and well-maintained, and is there a policy for > preventing cross-contamination of paraffin sections in the bath? > > I have a procedure for cutting the sections and cleaning water baths but I > guess I am not specifically using the word cross-contamination. Would > appreciate how other labs are "wording" this procedure. > > Thanks in advance!! > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SBarnes <@t> elch.org Tue Oct 26 06:27:32 2004 From: SBarnes <@t> elch.org (Sue Barnes) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Formula 83, xylene replacement Message-ID: > -----Original Message----- > From: Sue Barnes > Sent: Tuesday, October 26, 2004 7:04 AM > To: Histonet (E-mail) > Subject: Formula 83, xylene replacement > > Need information on formula 83. > > Is there anyone out there using it in place of xylene > Do you use it on the processor and on the stainer? > > Please contact me if you don't mind more questions. > > Thanks > Sue Barnes > East Liverpool City Hospital > East Liverpool, Ohio From amarusk1 <@t> FAIRVIEW.ORG Tue Oct 26 08:13:33 2004 From: amarusk1 <@t> FAIRVIEW.ORG (ANN MARUSKA) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] CD15 Message-ID: Hi Histonetters, We have recently had problems where our CD15 is NOT staining in any of the Hodgkins cases we had - although, the CD30 is staining quite well. Our control will stain well but we have had 3 patients who have stained negative with our procedure and yet positive at another institution. This has been on both bone marrow cores and nodes. We currently use Ventana's pre-dilute (clone MMA) on our Benchmark. Has this happened to anyone else - or do you have any suggestions? Thanks in advance. Ann Ann Maruska Fairview-University Medical Center Mpls. MN 55454 amarusk1@fairview.org 612-273-9119 The information transmitted in this e-mail is intended only for the person or entity to which it is addressed and may contain confidential and/or privileged material, including “protected health information.” If you are not the intended recipient, you are hereby notified that any review, retransmission, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. <<<>>> -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:ANN MARUSKA N:MARUSKA;ANN TEL;WORK:612-273-9119 ORG:;LAB EMAIL;WORK;PREF;NGW:AMARUSK1@FAIRVIEW.ORG END:VCARD From Michael.Rice <@t> holy-cross.com Tue Oct 26 08:22:02 2004 From: Michael.Rice <@t> holy-cross.com (Rice, Michael) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Looking for a Cryostat for Mohs Message-ID: <3BC92F29BE821745AB15E04C98EE028D69362B@HCH2KMAIL.holy-cross.com> I have used Leica cryostats for years for Mohs and they are great, push button feed and retract, hold a lot of specimens holders, maintain temperature well, and are comfortable to work on Mike Rice Holy Cross Hospital Ft Lauderdale, Fl -----Original Message----- From: ClarkatSCSC@aol.com [mailto:ClarkatSCSC@aol.com] Sent: Tuesday, October 26, 2004 12:27 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Looking for a Cryostat for Mohs Hi Everyone, I'm looking for a great cryostat for Mohs. I'm currently using a Tissue Tek thats about 30 yrs old and love it. I have tried a couple new ones on the market but nothing can compare. Does anyone feel that they have the best cryostat out there for Mohs. I would love to hear from you. Thanks. David Clark Portland, Or _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- From JNocito <@t> Pathreflab.com Tue Oct 26 08:45:56 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Working with toenails Message-ID: Hey everyone, short story first. When I was in high school, I was a running back in football (American football, not soccer). I quit because the middle line backers and defensive ends were twice my size and I was getting killed. Stay with me just a bit longer. I promise this will be worth it. I didn't realize that I could have been a place kicker or a punter. I could have been known as "Joe the Toe". Now, we are becoming the south Texas lab known for toenails, but we are having a problem with them staying on. We also perform a PAS/Fungus. We have tried Nair and ammonia water, but we are still having problems. Our sales manager is going to all the podiatrists in town and we are getting inundated with toenails. Can anyone give me some suggestions? This isn't what I had in mind when I said I wanted to be known as "Joe the Toe". thanks in advance. Joe "The Toe" Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX From Bauer.Karen <@t> mayo.edu Tue Oct 26 08:47:17 2004 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Looking for a Cryostat for Mohs Message-ID: I agree with Mike Rice. We have 2 Leica cryostats and they both work wonderfully. We do not use them just for Mohs. They cut all tissues well, including the many skin excisions that we receive. Karen Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI -----Original Message----- From: ClarkatSCSC@aol.com [mailto:ClarkatSCSC@aol.com] Sent: Monday, October 25, 2004 11:27 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Looking for a Cryostat for Mohs Hi Everyone, I'm looking for a great cryostat for Mohs. I'm currently using a Tissue Tek thats about 30 yrs old and love it. I have tried a couple new ones on the market but nothing can compare. Does anyone feel that they have the best cryostat out there for Mohs. I would love to hear from you. Thanks. David Clark Portland, Or _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. From HornHV <@t> archildrens.org Tue Oct 26 08:50:13 2004 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Looking for a Cryostat for Mohs Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6322B97F@EMAIL.archildrens.org> Yes we love our Leica cryostats. Very dependable, easy to use. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bauer, Karen Sent: Tuesday, October 26, 2004 8:47 AM To: 'ClarkatSCSC@aol.com'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Looking for a Cryostat for Mohs I agree with Mike Rice. We have 2 Leica cryostats and they both work wonderfully. We do not use them just for Mohs. They cut all tissues well, including the many skin excisions that we receive. Karen Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI -----Original Message----- From: ClarkatSCSC@aol.com [mailto:ClarkatSCSC@aol.com] Sent: Monday, October 25, 2004 11:27 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Looking for a Cryostat for Mohs Hi Everyone, I'm looking for a great cryostat for Mohs. I'm currently using a Tissue Tek thats about 30 yrs old and love it. I have tried a couple new ones on the market but nothing can compare. Does anyone feel that they have the best cryostat out there for Mohs. I would love to hear from you. Thanks. David Clark Portland, Or _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From David.Edmondson <@t> christie-tr.nwest.nhs.uk Tue Oct 26 08:54:12 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Working with toenails Message-ID: Was a 2% Phenol solution another softening up option? Dave Christie Hosp Manchester UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joe Nocito Sent: 26 October 2004 14:46 To: Histonet Subject: [Histonet] Working with toenails Hey everyone, short story first. When I was in high school, I was a running back in football (American football, not soccer). I quit because the middle line backers and defensive ends were twice my size and I was getting killed. Stay with me just a bit longer. I promise this will be worth it. I didn't realize that I could have been a place kicker or a punter. I could have been known as "Joe the Toe". Now, we are becoming the south Texas lab known for toenails, but we are having a problem with them staying on. We also perform a PAS/Fungus. We have tried Nair and ammonia water, but we are still having problems. Our sales manager is going to all the podiatrists in town and we are getting inundated with toenails. Can anyone give me some suggestions? This isn't what I had in mind when I said I wanted to be known as "Joe the Toe". thanks in advance. Joe "The Toe" Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Bauer.Karen <@t> mayo.edu Tue Oct 26 08:56:47 2004 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Working with toenails Message-ID: Joe, We have had many problems with our toenails and fingernails staying on our slides. Lately we have been having good luck with them. We do not soak or pretreat the nails before processing, but once they are embedded and then faced into, we soak the block in soapy water. (A few drops of Dawn dish soap in a weigh boat with about 10 to 20 cc of tap water) Let it soak for a while, cut a section and place it on a slide. We have had good luck with Erie Scientifics Superfrost Plus Gold slides. So far the sections have stayed on pretty well. We also use regular charged slides with a bit of Elmer's Glue smeared on them. Put the section on and air dry for about 30 minutes. This works well too, but there is background from the glue. Good Luck, I know they are a pain to keep on the slides. Karen Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI -----Original Message----- From: Joe Nocito [mailto:JNocito@Pathreflab.com] Sent: Tuesday, October 26, 2004 8:46 AM To: Histonet Subject: [Histonet] Working with toenails Hey everyone, short story first. When I was in high school, I was a running back in football (American football, not soccer). I quit because the middle line backers and defensive ends were twice my size and I was getting killed. Stay with me just a bit longer. I promise this will be worth it. I didn't realize that I could have been a place kicker or a punter. I could have been known as "Joe the Toe". Now, we are becoming the south Texas lab known for toenails, but we are having a problem with them staying on. We also perform a PAS/Fungus. We have tried Nair and ammonia water, but we are still having problems. Our sales manager is going to all the podiatrists in town and we are getting inundated with toenails. Can anyone give me some suggestions? This isn't what I had in mind when I said I wanted to be known as "Joe the Toe". thanks in advance. Joe "The Toe" Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. From TMcNemar <@t> lmhealth.org Tue Oct 26 09:12:53 2004 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Formalin substitutes... Message-ID: <90092A4ED388D7119575006008F7112049CBEA@NT_EXCHANGE> Hello all, It's been a number of years since we looked at the substitutes. I was wondering how good they are these days. Is anybody using one? Any recommendations? I would appreciat any comments. Thanks. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 From jcox90 <@t> yahoo.com Tue Oct 26 09:21:27 2004 From: jcox90 <@t> yahoo.com (Jill Cox) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] New microtomy website is fixed!!! Message-ID: <20041026142127.14674.qmail@web40908.mail.yahoo.com> Hello Netters, I want to thank you all for finding my typo!! I probably didn't see it because it was right in front of me. If anyone needs control slides just let me know. I will be providing free shipping until the end of the year if you let me know you are from the Histonet...Thank you again for all the feedback and comments about starting a company, I couldn't have done this without you!!! You can visit www.precisionmicrotomy.com if you need more info. I will be getting a brochure together in the near future also. Have a great day, Jill Jill Cox HT (ASCP) --------------------------------- Do you Yahoo!? Yahoo! Mail Address AutoComplete - You start. We finish. From JNocito <@t> Pathreflab.com Tue Oct 26 09:41:04 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Working with toenails In-Reply-To: Message-ID: See, you're the greatest group I've have the honor of knowing. I've received so many replies from pre-soaking in Nair before processing, to using soapy water to a 2% phenol solution. I appreciate everyone's input. Thanks again. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joe Nocito Sent: Tuesday, October 26, 2004 8:46 AM To: Histonet Subject: [Histonet] Working with toenails Hey everyone, short story first. When I was in high school, I was a running back in football (American football, not soccer). I quit because the middle line backers and defensive ends were twice my size and I was getting killed. Stay with me just a bit longer. I promise this will be worth it. I didn't realize that I could have been a place kicker or a punter. I could have been known as "Joe the Toe". Now, we are becoming the south Texas lab known for toenails, but we are having a problem with them staying on. We also perform a PAS/Fungus. We have tried Nair and ammonia water, but we are still having problems. Our sales manager is going to all the podiatrists in town and we are getting inundated with toenails. Can anyone give me some suggestions? This isn't what I had in mind when I said I wanted to be known as "Joe the Toe". thanks in advance. Joe "The Toe" Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MAUGER <@t> email.chop.edu Tue Oct 26 10:12:52 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] eosin Message-ID: Thanks for all the varied answers about eosin in etoh on the processor! We are trying your suggestions to find whats best for us. Jo >>> "Joe Nocito" 10/26/04 10:41AM >>> See, you're the greatest group I've have the honor of knowing. I've received so many replies from pre-soaking in Nair before processing, to using soapy water to a 2% phenol solution. I appreciate everyone's input. Thanks again. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joe Nocito Sent: Tuesday, October 26, 2004 8:46 AM To: Histonet Subject: [Histonet] Working with toenails Hey everyone, short story first. When I was in high school, I was a running back in football (American football, not soccer). I quit because the middle line backers and defensive ends were twice my size and I was getting killed. Stay with me just a bit longer. I promise this will be worth it. I didn't realize that I could have been a place kicker or a punter. I could have been known as "Joe the Toe". Now, we are becoming the south Texas lab known for toenails, but we are having a problem with them staying on. We also perform a PAS/Fungus. We have tried Nair and ammonia water, but we are still having problems. Our sales manager is going to all the podiatrists in town and we are getting inundated with toenails. Can anyone give me some suggestions? This isn't what I had in mind when I said I wanted to be known as "Joe the Toe". thanks in advance. Joe "The Toe" Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ 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 From laurie.colbert <@t> huntingtonhospital.com Tue Oct 26 10:19:44 2004 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Tissue Capture Pen Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5C0DA@EXCHANGE1.huntingtonhospital.com> Has anyone tried the Tissue Capture Pen from Cancer Diagnostics for keeping tissues sections from falling off of the slides? Does it work?? Laurie Colbert From vazquezr <@t> ohsu.edu Tue Oct 26 10:23:26 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Looking for a Cryostat for Mohs Message-ID: Dave, I have a tissue -tek over here in derm surg that I am not using. Would have to talk to Nancy though. Robyn Derm Surg Portland, OR >>> 10/25/2004 9:26:46 PM >>> Hi Everyone, I'm looking for a great cryostat for Mohs. I'm currently using a Tissue Tek thats about 30 yrs old and love it. I have tried a couple new ones on the market but nothing can compare. Does anyone feel that they have the best cryostat out there for Mohs. I would love to hear from you. Thanks. David Clark Portland, Or _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Mandy <@t> serotec.co.uk Tue Oct 26 10:41:06 2004 From: Mandy <@t> serotec.co.uk (Mandy Townsend) Date: Fri Sep 16 15:24:12 2005 Subject: [Histonet] Re: Automated vs Manual Message-ID: Dear All We often experience that when using some antibodies in an automated IHC system that they require different pre-treatment requirements than they do if using in a manual system. Do other people see this, perhaps there has already been a discussion on histonet concerning this that someone could direct me to. Thanks for your help. Mandy From jcox90 <@t> yahoo.com Tue Oct 26 10:42:29 2004 From: jcox90 <@t> yahoo.com (Jill Cox) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Looking for a Cryostat for Mohs In-Reply-To: Message-ID: <20041026154229.52089.qmail@web40911.mail.yahoo.com> Dave, You can also try www.labx.com. They have a lot of cryostats that are very inexpensive. I have also found some on ebay. There is a lot of reputable companies on ebay just make sure you check them out. Hope this helps, jill Robyn Vazquez wrote: Dave, I have a tissue -tek over here in derm surg that I am not using. Would have to talk to Nancy though. Robyn Derm Surg Portland, OR >>> 10/25/2004 9:26:46 PM >>> Hi Everyone, I'm looking for a great cryostat for Mohs. I'm currently using a Tissue Tek thats about 30 yrs old and love it. I have tried a couple new ones on the market but nothing can compare. Does anyone feel that they have the best cryostat out there for Mohs. I would love to hear from you. Thanks. David Clark Portland, Or _______________________________________________ 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 Jill Cox HT (ASCP) --------------------------------- Do you Yahoo!? Yahoo! Mail Address AutoComplete - You start. We finish. From TJJ <@t> Stowers-Institute.org Tue Oct 26 11:28:49 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Bone suture staining Message-ID: Tricia, I ask questions a lot and don't get answers. Either we're trying stuff nobody does, trying something someone else wants to publish, or perhaps the person with the answer has temporarily unsubscribed to avoid the dreaded "out of office" loop! At any rate, have you thought of doing a trichrome stain? Or maybe an iron hematoxylin counterstained with alcoholic saffron or metanil yellow? I don't know if that would pick it up, but if you have some on hand, drop a slide in and see what you get. Bummer is the bone usually will stain the same color as the connective tissue.... Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From funderwood <@t> mcohio.org Tue Oct 26 11:41:36 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:13 2005 Subject: [BULK] - [Histonet] Working with toenails Message-ID: Liquid fabric softener works well. My ice tray is actually a frozen block of 0.2% fabric softener. I have mentioned this before, but I have found titebond (a commercial wood glue, use a 5% solution) is the "bomb" for adhering hard tissues to slides. Fred >>> "Joe Nocito" 10/26/04 09:45AM >>> Hey everyone, short story first. When I was in high school, I was a running back in football (American football, not soccer). I quit because the middle line backers and defensive ends were twice my size and I was getting killed. Stay with me just a bit longer. I promise this will be worth it. I didn't realize that I could have been a place kicker or a punter. I could have been known as "Joe the Toe". Now, we are becoming the south Texas lab known for toenails, but we are having a problem with them staying on. We also perform a PAS/Fungus. We have tried Nair and ammonia water, but we are still having problems. Our sales manager is going to all the podiatrists in town and we are getting inundated with toenails. Can anyone give me some suggestions? This isn't what I had in mind when I said I wanted to be known as "Joe the Toe". thanks in advance. Joe "The Toe" Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcline <@t> wchsys.org Tue Oct 26 11:59:26 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Joe the Toe Message-ID: <001901c4bb7d$270df9f0$1d2a14ac@wchsys.org> We have smeared the slides with Sta-on solution, and let them dry a little before mounting section of toenail. I use this in our waterbaths for adhesion. Sta-on is bought from Surgipath ***** CONFIDENTIALITY NOTICE ***** This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. From Jamie.Dukes <@t> se.amedd.army.mil Tue Oct 26 13:26:19 2004 From: Jamie.Dukes <@t> se.amedd.army.mil (Dukes, Jamie Mr EAMC) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Lymph node solution Message-ID: Please help me with instruction on using lymph node solution. From Jamie.Dukes <@t> se.amedd.army.mil Tue Oct 26 13:28:49 2004 From: Jamie.Dukes <@t> se.amedd.army.mil (Dukes, Jamie Mr EAMC) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] FW: Lymph node solution Message-ID: ________________________________ From: Dukes, Jamie Mr EAMC Sent: Tuesday, October 26, 2004 2:26 PM To: 'histonet@pathology.swmed.EDU' Subject: Lymph node solution Please help me with instruction on using lymph node solution. From funderwood <@t> mcohio.org Tue Oct 26 13:51:49 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:13 2005 Subject: [BULK] - [Histonet] Working with toenails Message-ID: Hi Fran, I use it for all tissues. Some samples I limit the exposure, for example neural tissue. Don't be tempted to make a stronger (5% and greater) solution. Results are less than satisfactory. It does help though on hard or bloody tissue. Fred >>> "Fran Lemons" 10/26/04 02:05PM >>> Do you use the fabric softener 'brick' for all of your tissues, or just nails? >>> "Fred Underwood" 10/26/04 12:41PM >>> Liquid fabric softener works well. My ice tray is actually a frozen block of 0.2% fabric softener. I have mentioned this before, but I have found titebond (a commercial wood glue, use a 5% solution) is the "bomb" for adhering hard tissues to slides. Fred >>> "Joe Nocito" 10/26/04 09:45AM >>> Hey everyone, short story first. When I was in high school, I was a running back in football (American football, not soccer). I quit because the middle line backers and defensive ends were twice my size and I was getting killed. Stay with me just a bit longer. I promise this will be worth it. I didn't realize that I could have been a place kicker or a punter. I could have been known as "Joe the Toe". Now, we are becoming the south Texas lab known for toenails, but we are having a problem with them staying on. We also perform a PAS/Fungus. We have tried Nair and ammonia water, but we are still having problems. Our sales manager is going to all the podiatrists in town and we are getting inundated with toenails. Can anyone give me some suggestions? This isn't what I had in mind when I said I wanted to be known as "Joe the Toe". thanks in advance. Joe "The Toe" Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ 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 From weneng2004 <@t> yahoo.com Tue Oct 26 14:00:56 2004 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] human Ab on human tumor Message-ID: <20041026190056.18665.qmail@web53408.mail.yahoo.com> Hi histonetters, Currently I am doing IHC using human Ab on human tumor. I know it would cause "messy" background. But this tumor was developed by injection human tumor cell to nude mice. The results came out pretty good. I didn't use conjugated primary Ab, but just traditional method. I think it's because the tumor was developed in mice, the tissue would not contain endogenous human serum that causes the "messy background", although the tumor is human tumor. Am I right on this? Or my stain result just "fake"? Hope I said it clearly. Could any expert give me help? Thanks in advance! --------------------------------- Do you Yahoo!? Read only the mail you want - Yahoo! Mail SpamGuard. From twheelock <@t> mclean.harvard.edu Tue Oct 26 14:43:42 2004 From: twheelock <@t> mclean.harvard.edu (Timothy R. Wheelock) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] IMMUNOSTAIN OUT-SOURCING Message-ID: <417EA8EE.5060608@mclean.harvard.edu> Hi Everyone: I was wondering if anyone knew of a laboratory or company that did immunostaining for a fee. I could cut the paraffin sections, ship them to the lab, and then receive the immuno-stained slides back. We would only need a total of about 140 slides per year. The lab would need to be able to do the following stains: Ubiquitin Alpha-synuclein Huntingtin Phosphorylated tau (AT-8) GFAP Calbindin Beta-amyloid HLA-DR. These are all for neuropathological diagnosis. Thank you, Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital Belmont, MA Any information, including protected health information (PHI), transmitted in this email is intended only for the person or entity to which it is addressed and may contain information that is privileged, confidential and or exempt from disclosure under applicable Federal or State law. Any review, retransmission, dissemination or other use of or taking of any action in reliance upon, protected health information (PHI) by persons or entities other than the intended recipient is prohibited. If you received this email in error, please contact the sender and delete the material from any computer. From ihc <@t> unipathllc.com Tue Oct 26 15:48:47 2004 From: ihc <@t> unipathllc.com (UniPath IHC) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Eosinophil marker Message-ID: <000601c4bb9d$31220830$4500a8c0@unipathllc.corp> Can anyone recommend an eosinophil antibody that works in FFPE tissue? From vazquezr <@t> ohsu.edu Tue Oct 26 16:14:07 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:13 2005 Subject: [BULK] - [Histonet] Working with toenails Message-ID: Fred, Titebond would work on frozen tissue as well? Robyn OHSU >>> "Fred Underwood" 10/26/2004 9:41:36 AM >>> Liquid fabric softener works well. My ice tray is actually a frozen block of 0.2% fabric softener. I have mentioned this before, but I have found titebond (a commercial wood glue, use a 5% solution) is the "bomb" for adhering hard tissues to slides. Fred >>> "Joe Nocito" 10/26/04 09:45AM >>> Hey everyone, short story first. When I was in high school, I was a running back in football (American football, not soccer). I quit because the middle line backers and defensive ends were twice my size and I was getting killed. Stay with me just a bit longer. I promise this will be worth it. I didn't realize that I could have been a place kicker or a punter. I could have been known as "Joe the Toe". Now, we are becoming the south Texas lab known for toenails, but we are having a problem with them staying on. We also perform a PAS/Fungus. We have tried Nair and ammonia water, but we are still having problems. Our sales manager is going to all the podiatrists in town and we are getting inundated with toenails. Can anyone give me some suggestions? This isn't what I had in mind when I said I wanted to be known as "Joe the Toe". thanks in advance. Joe "The Toe" Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ 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 From tahseen <@t> brain.net.pk Tue Oct 26 17:42:00 2004 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Looking for a Cryostat for Mohs References: <30.63e551b3.2eaf2c06@aol.com> Message-ID: <003201c4bbad$02a32880$972bfea9@m7c0y4> Yes we love our Leica cryostats. Very dependable, easy to use. Muhammad Tahseen Histology Supervisor SKMCH & RC Lahore Pakistan ----- Original Message ----- From: To: Sent: Tuesday, October 26, 2004 9:26 AM Subject: [Histonet] Looking for a Cryostat for Mohs > Hi Everyone, > > I'm looking for a great cryostat for Mohs. I'm currently using a Tissue Tek > thats about 30 yrs old and love it. I have tried a couple new ones on the > market but nothing can compare. Does anyone feel that they have the best cryostat > out there for Mohs. I would love to hear from you. > > Thanks. > > David Clark > Portland, Or > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From hasselblatt <@t> uni-muenster.de Tue Oct 26 19:16:29 2004 From: hasselblatt <@t> uni-muenster.de (Dr. med. Martin Hasselblatt) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Plasma Cell Antibodies? Message-ID: Dear Histonet-Members, I am searching for an specific and reliable plasma cell marker for routine diagnostics (paraffin-embedded tissue). We currently use antibodies from DAKO (kappa light chain, lambda light chain and VS38c), but are not so happy with the specificity of VS38c. What plasma cell markers are you using out there? Similiar experiences with VS38c? Thank you very much for your response! Best regards, Martin ______________________________________ Martin Hasselblatt MD Institute of Neuropathology University Hospital M?nster, Germany From AnthonyH <@t> chw.edu.au Tue Oct 26 19:30:28 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] CD15 Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E0E7@simba.kids> Ann, CD15 antibodies a very finicky. Being an IgM antibody, you might need to use a anti-mouse IgM after the localisation antibody to improve the sensitivity. I have also found a difference in clones with Leu-M1 from Becton-Dickinson better than Dako's C3D-1. Often overnight incubation with the Dako antibody will also improve the sensitivity. Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: ANN MARUSKA [mailto:amarusk1@FAIRVIEW.ORG] Sent: Tuesday, 26 October 2004 11:14 PM To: histonet@pathology.swmed.EDU Subject: [Histonet] CD15 Hi Histonetters, We have recently had problems where our CD15 is NOT staining in any of the Hodgkins cases we had - although, the CD30 is staining quite well. Our control will stain well but we have had 3 patients who have stained negative with our procedure and yet positive at another institution. This has been on both bone marrow cores and nodes. We currently use Ventana's pre-dilute (clone MMA) on our Benchmark. Has this happened to anyone else - or do you have any suggestions? Thanks in advance. Ann Ann Maruska Fairview-University Medical Center Mpls. MN 55454 amarusk1@fairview.org 612-273-9119 The information transmitted in this e-mail is intended only for the person or entity to which it is addressed and may contain confidential and/or privileged material, including "protected health information." If you are not the intended recipient, you are hereby notified that any review, retransmission, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. <<<>>> ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From hlukey <@t> msn.com Tue Oct 26 19:41:09 2004 From: hlukey <@t> msn.com (Hugh Luk) Date: Fri Sep 16 15:24:13 2005 Subject: [BULK] - [Histonet] Working with toenails Message-ID: Joe, I worked with Ben Manuel formerly of AFIP, and he did a two step addition. Sometimes toenails can get kind of thick and the chitin needs softening (diabetes ect.), so we used the AFIP (green book-it's not in the new one) recipe for chitin softening, but we modified it to use Zinc Chloride instead of Mercuric Chloride. We soaked the toenails after grossing it. Several hours normally did well. The H&E and PAS-f were not compromised. Ben used to cut his sections on re-treated adhesive slides (we buy "Plus" slides, then coat them again with diluted poly-L-lysine from Sigma) and he would place his slides in a forced air dryer flat, with A SEMI-WET PAPER TOWEL ON TOP OF THE SLIDES. When the paper towel and the slides were dry, we deparaffinized them ect. If the toenail still started falling off, we would not dilute the Sigma solution and coat the slide using a cotton tip applicator, let it dry, and rock-on. If microtomy was still an issue, face the block, and soften the face with "Chitin softening solution." We pour it into a screw-cap cup, then wash it and cool it again on ice. I've heard of Fred's suggestion from several techs, so I'm sure it works well too. However, because we cut our IHC on the same ice, we don't add anything to the water. We just haven't tested it. Zinc chloride makes administrators cringe, and the paper towel in an oven could be problematic. Good luck with your toenails. We hate them! And that was an awesome set-up for a question. >From: "Fred Underwood" >To: , >Subject: Re: [BULK] - [Histonet] Working with toenails >Date: Tue, 26 Oct 2004 12:41:36 -0400 > >Liquid fabric softener works well. My ice tray is actually a frozen >block of 0.2% fabric softener. I have mentioned this before, but I have >found titebond (a commercial wood glue, use a 5% solution) is the "bomb" >for adhering hard tissues to slides. > >Fred > > >>> "Joe Nocito" 10/26/04 09:45AM >>> >Hey everyone, >short story first. When I was in high school, I was a running back in >football (American football, not soccer). I quit because the middle >line >backers and defensive ends were twice my size and I was getting >killed. > Stay with me just a bit longer. I promise this will be worth >it. > I didn't realize that I could have been a place kicker or a >punter. I could >have been known as "Joe the Toe". >Now, we are becoming the south Texas lab known for toenails, but we >are >having a problem with them staying on. We also perform a PAS/Fungus. We >have >tried Nair and ammonia water, but we are still having problems. Our >sales >manager is going to all the podiatrists in town and we are getting >inundated >with toenails. Can anyone give me some suggestions? > This isn't what I had in mind when I said I wanted to be known >as "Joe the >Toe". > thanks in advance. > > >Joe "The Toe" Nocito, BS, HT(ASCP) QIHC >Histology Manager >Pathology Reference Lab >San Antonio, TX > > >_______________________________________________ >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 From gu.lang <@t> gmx.at Wed Oct 27 06:06:40 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] mohs' technique Message-ID: <005001c4bc15$09cef630$eeeea8c0@server> Can anyone explain me shortly this special thing? Gudrun From rentonlf <@t> bru.wits.ac.za Wed Oct 27 05:57:23 2004 From: rentonlf <@t> bru.wits.ac.za (renton louise mrs) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] ?titebond Message-ID: <1098874643.8c61d4c0rentonlf@bru.wits.ac.za> Fred, assuming that titebond is available in this country, how do you use it on your slides? smear, dip, drop or what? What's the background like? If you could I would appreciate you sharing your protocol with me best regards Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? From i_stain <@t> yahoo.com Wed Oct 27 06:13:30 2004 From: i_stain <@t> yahoo.com (Scott Mordue) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] CD52 Message-ID: <20041027111331.34875.qmail@web42008.mail.yahoo.com> Hi, Looking for a good antibody for CD52 on FFPE human tissue. If anyone know the source to buy this antibody as well as the protocol, please help. Thanks Scott --------------------------------- Do you Yahoo!? Yahoo! Mail Address AutoComplete - You start. We finish. From MAUGER <@t> email.chop.edu Wed Oct 27 06:55:54 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] IMMUNOSTAIN OUT-SOURCING Message-ID: Hi Tim, I know of a great lab for out sourcing; MDR Global Systems in Windber, PA. They offer excellent services. The # is 814-467-9111(ofc.) 814-467-9112(lab) or you can email Alan at ihcguy@aol.com. Jo Mauger >>> "Timothy R. Wheelock" 10/26/04 03:43PM >>> Hi Everyone: I was wondering if anyone knew of a laboratory or company that did immunostaining for a fee. I could cut the paraffin sections, ship them to the lab, and then receive the immuno-stained slides back. We would only need a total of about 140 slides per year. The lab would need to be able to do the following stains: Ubiquitin Alpha-synuclein Huntingtin Phosphorylated tau (AT-8) GFAP Calbindin Beta-amyloid HLA-DR. These are all for neuropathological diagnosis. Thank you, Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital Belmont, MA Any information, including protected health information (PHI), transmitted in this email is intended only for the person or entity to which it is addressed and may contain information that is privileged, confidential and or exempt from disclosure under applicable Federal or State law. Any review, retransmission, dissemination or other use of or taking of any action in reliance upon, protected health information (PHI) by persons or entities other than the intended recipient is prohibited. If you received this email in error, please contact the sender and delete the material from any computer. From MAUGER <@t> email.chop.edu Wed Oct 27 07:07:18 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] CD52 Message-ID: Hi Scott, I found a good CD52 from Caltag Labs #MHCD5200. It's a ms monoclonal IgG. Works well with heat retrieval in EDTA buffer (pH 6.) Good luck. Jo >>> "Scott Mordue" 10/27/04 07:13AM >>> Hi, Looking for a good antibody for CD52 on FFPE human tissue. If anyone know the source to buy this antibody as well as the protocol, please help. Thanks Scott --------------------------------- Do you Yahoo!? Yahoo! Mail Address AutoComplete - You start. We finish. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ajennings <@t> unmc.edu Wed Oct 27 08:23:03 2004 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] human Ab on human tumor In-Reply-To: <20041026190056.18665.qmail@web53408.mail.yahoo.com> Message-ID: clarification please your routine immuno includes ? looking for human cells in mouse? your primary antibody is made against human tumor cells, what animal is the antibody made in? what do you use as a secondary antibody? anita wen eng To Sent by: histonet histonet-bounces@ lists.utsouthwest cc ern.edu Subject [Histonet] human Ab on human tumor 10/26/2004 02:00 PM Hi histonetters, Currently I am doing IHC using human Ab on human tumor. I know it would cause "messy" background. But this tumor was developed by injection human tumor cell to nude mice. The results came out pretty good. I didn't use conjugated primary Ab, but just traditional method. I think it's because the tumor was developed in mice, the tissue would not contain endogenous human serum that causes the "messy background", although the tumor is human tumor. Am I right on this? Or my stain result just "fake"? Hope I said it clearly. Could any expert give me help? Thanks in advance! --------------------------------- Do you Yahoo!? Read only the mail you want - Yahoo! Mail SpamGuard. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RCazares <@t> schosp.org Wed Oct 27 09:29:56 2004 From: RCazares <@t> schosp.org (Cazares, Ruth) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Bone marrow core biopsy handling Message-ID: <913FAC2B773C19488E26AE6572180FA53AF915@exch01.schosp.org> Hello again, I am interested to know how bone marrow cores are handled out there in histo-land. In our lab we put the cores in a formic acid decalcifier (which is supposed to fix the tissue as it decals according to the company) as soon as we get it. It stays in for 2-3 hours depending on the amount of bone it contains. Then it is processed on the routine overnight program for large tissues. We do not use a short program for BM cores. The H&E's are ok, I guess, but nothing compared to B-5 fixation. We have gotten rid of all the mercury in our labs and I'm wondering what are the alternatives that people are using? Thanks again! Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. From jkiernan <@t> uwo.ca Wed Oct 27 10:27:55 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Eosinophil marker References: <000601c4bb9d$31220830$4500a8c0@unipathllc.corp> Message-ID: <417FBE7B.E25670D1@uwo.ca> You identify yourself only as "Unipath IHC." Why? Are you a commercial outfit asking for free advice on the internet? If so, would you sell services to clients on the strength of someone's 15-minute response on a listserver? Internet replies, including mine (which follows) have no real value. If you do work for a client based on advice collected from an internet enquiry you are asking to be sued. That said, I'll offer some advice. do not follow it without checking that it's based on established and generally accepted knowledge. Compare 3 major textbooks. If there are discrepancies, follow up the references in your library. Get photocopies of older papers by interlibrary loan if necessary. That's one of the ways to get details of techniques. Antibodies are expensive. Eosin isn't, and it stains the cytoplasmic granules of eosinophils strongly. If you stain with an alkaline (pH 9) aqueous solution of eosin, the only red things in animal tissues will be eosinophil granules, Paneth cell granules (in the intestine) and tails of spermatozoa (if present). I think this is a complete list; someone, please correct me if I've missed something. If you need a counterstain, use any haemalum (Ehrlich, Gill, Harris, Mayer etc) or a blue basic dye (eg toluidine blue at pH 4) according to your needs. Either of these "counterstains" should probably be done before the alkaline eosin staining. Dyes are cheaper and easier to use than antibodies! Immunohistochemistry is an important family of methods, but it is expensive and is not needed for simple jobs such as staining eosinophils. Your enquiry did include information about the tissue you work with. Whatever it is, you should not confuse eosinophils with Paneth cells or sperm tails! -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ UniPath IHC wrote: > > Can anyone recommend an eosinophil antibody that works in FFPE tissue? > > _______________________________________________ From am102 <@t> st-andrews.ac.uk Wed Oct 27 11:00:58 2004 From: am102 <@t> st-andrews.ac.uk (Anne-Sophie Martinez) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] (no subject) Message-ID: <200410271546.QAA06358@bute.st-andrews.ac.uk> Hi everybody, I am trying to stain in IHC, 2 isoforms of a protein present in the kidney tubules of some fish. I would like to know if these proteins colocalise or not in the same tubules. I can't use any confocal microscopy because I use two specific polyclonal antibodies, both raised in rabbit. My option is then to have serial sections that I will stain: one with AB1 with a 2dry AB-FITC and one with AB2 with a 2dry AB-Cy3. But this procedure bring two problems: (i) even serial sections don't have exactly the same structure and (ii) it is difficult to localise the same tubules on two different slides. Hope all of this is clear. Would somebody have a suggestion for a better procedure or how to improve this one? Thanks. AS ---------------------------------------------- Dr Anne-Sophie Martinez School of Biology, University of St. Andrews, Bute Medical Buildings, St. Andrews, KY16 9TS, UK. Tel +44 (0) 1334 463063/3365 Fax +44 (0) 1334 463600 ---------------------------------------------- From rocan <@t> mac.com Wed Oct 27 11:22:15 2004 From: rocan <@t> mac.com (Rocan) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] (no subject) In-Reply-To: <200410271546.QAA06358@bute.st-andrews.ac.uk> References: <200410271546.QAA06358@bute.st-andrews.ac.uk> Message-ID: <5D573E92-2834-11D9-8F98-000A9589219E@mac.com> I would suggest using labeling kits for your antibodies. Molecular Probes has some that are very easy to use. You could directly label each antibody with a different alexa dye and have the result you want. This may cost a bit but you can label several antibodies even make your own secondaries for other applications. I believe the kist are called Zenon. Good luck Rocio ----- Dr.Rocio Sierra-Honigmann Director Engineered Wound Repair Laboratory Cedars Sinai Research Institute Davis 1091 310-423-1882 Honigmannr@cshs.org k On Oct 27, 2004, at 9:00 AM, Anne-Sophie Martinez wrote: > Hi everybody, > > I am trying to stain in IHC, 2 isoforms of a protein present in the > kidney > tubules of some fish. I would like to know if these proteins > colocalise or > not in the same tubules. > I can't use any confocal microscopy because I use two specific > polyclonal > antibodies, both raised in rabbit. My option is then to have serial > sections that I will stain: one with AB1 with a 2dry AB-FITC and one > with > AB2 with a 2dry AB-Cy3. But this procedure bring two problems: (i) even > serial sections don't have exactly the same structure and (ii) it is > difficult to localise the same tubules on two different slides. Hope > all of > this is clear. > Would somebody have a suggestion for a better procedure or how to > improve > this one? > Thanks. > > AS > > > ---------------------------------------------- > Dr Anne-Sophie Martinez > School of Biology, > University of St. Andrews, > Bute Medical Buildings, > St. Andrews, KY16 9TS, UK. > > Tel +44 (0) 1334 463063/3365 > Fax +44 (0) 1334 463600 > ---------------------------------------------- > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Wed Oct 27 11:23:44 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Looking for Charles Embry Message-ID: Mr. Embry, please contact me via email or telephone (210) 231-8058. I will be here until 4:00 PM CDT. Thank you. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX From ihc <@t> unipathllc.com Wed Oct 27 11:34:50 2004 From: ihc <@t> unipathllc.com (UniPath IHC) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Eosinophil marker In-Reply-To: <417FBE7B.E25670D1@uwo.ca> Message-ID: <000001c4bc42$e21aff20$4500a8c0@unipathllc.corp> I guess I should have made myself clear, and I admit I forgot to sign my name before I sent off the email. I'm an IHC tech working for a private company. However, I'm not just trying to get "free advice on the internet". I have been a member of the Histonet community for four years and have enjoyed reading and contributing to this service. I would never use a response from Histonet as anything but advice or a hint in the right direction. As for who I work for, I have never been told that working for a private lab as opposed to a hospital excluded me from Histonet. I am working on a research project for a couple of doctors at the National Jewish in Denver. I am trying to doublestain eosinophils in skin biopsies by IHC and am having no luck with the MBP antibody (Chemicon) on routinely processed FFPE. It works fine for me on frozen sections, and even a little bit on tissue fixed in 4% PFA. I found a few emails in the archives regarding eosinophil markers, but most people asking what I need to know had never been replied to, at least not via the Histonet. I have considered special stains to mark eosinophils, but the doctors I am working with would like to use IHC. If I cannot make this antibody (MBP) work on FFPE then I need to find another eosinophil marker that does. Any help would be appreciated. Thanks for your time, Brianna Jackson, BS, QIHC UniPath, LLC Denver, CO 303-512-2220 bjackson@unipathllc.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Wednesday, October 27, 2004 9:28 AM To: UniPath IHC Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Eosinophil marker You identify yourself only as "Unipath IHC." Why? Are you a commercial outfit asking for free advice on the internet? If so, would you sell services to clients on the strength of someone's 15-minute response on a listserver? Internet replies, including mine (which follows) have no real value. If you do work for a client based on advice collected from an internet enquiry you are asking to be sued. That said, I'll offer some advice. do not follow it without checking that it's based on established and generally accepted knowledge. Compare 3 major textbooks. If there are discrepancies, follow up the references in your library. Get photocopies of older papers by interlibrary loan if necessary. That's one of the ways to get details of techniques. Antibodies are expensive. Eosin isn't, and it stains the cytoplasmic granules of eosinophils strongly. If you stain with an alkaline (pH 9) aqueous solution of eosin, the only red things in animal tissues will be eosinophil granules, Paneth cell granules (in the intestine) and tails of spermatozoa (if present). I think this is a complete list; someone, please correct me if I've missed something. If you need a counterstain, use any haemalum (Ehrlich, Gill, Harris, Mayer etc) or a blue basic dye (eg toluidine blue at pH 4) according to your needs. Either of these "counterstains" should probably be done before the alkaline eosin staining. Dyes are cheaper and easier to use than antibodies! Immunohistochemistry is an important family of methods, but it is expensive and is not needed for simple jobs such as staining eosinophils. Your enquiry did include information about the tissue you work with. Whatever it is, you should not confuse eosinophils with Paneth cells or sperm tails! -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ UniPath IHC wrote: > > Can anyone recommend an eosinophil antibody that works in FFPE tissue? > > _______________________________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Wed Oct 27 11:44:11 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:13 2005 Subject: [BULK] - [Histonet] Working with toenails Message-ID: I've not tried it on frozens. However, I have stained paraffin sections that have not been heat fixed, only air dried at room temperature. The sections stay on the slides suprisingly well. It is probably worth the effort to try it on frozen sections. Fred >>> "Robyn Vazquez" 10/26/04 05:14PM >>> Fred, Titebond would work on frozen tissue as well? Robyn OHSU >>> "Fred Underwood" 10/26/2004 9:41:36 AM >>> Liquid fabric softener works well. My ice tray is actually a frozen block of 0.2% fabric softener. I have mentioned this before, but I have found titebond (a commercial wood glue, use a 5% solution) is the "bomb" for adhering hard tissues to slides. Fred >>> "Joe Nocito" 10/26/04 09:45AM >>> Hey everyone, short story first. When I was in high school, I was a running back in football (American football, not soccer). I quit because the middle line backers and defensive ends were twice my size and I was getting killed. Stay with me just a bit longer. I promise this will be worth it. I didn't realize that I could have been a place kicker or a punter. I could have been known as "Joe the Toe". Now, we are becoming the south Texas lab known for toenails, but we are having a problem with them staying on. We also perform a PAS/Fungus. We have tried Nair and ammonia water, but we are still having problems. Our sales manager is going to all the podiatrists in town and we are getting inundated with toenails. Can anyone give me some suggestions? This isn't what I had in mind when I said I wanted to be known as "Joe the Toe". thanks in advance. Joe "The Toe" Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ 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 From MinHan.Tan <@t> vai.org Wed Oct 27 12:14:29 2004 From: MinHan.Tan <@t> vai.org (Tan, MinHan) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Any fixative to reduce background in immunofluorescence? Message-ID: Hi, I am preparing to do some immunofluorescence staining in kidney tissue with cd31. Some preliminary work in formalin fixed paraffin embedded tissue has yielded quite significant background fluorescence. A colleague suggested that I use frozen tissue, but the morphology is quite unacceptable. Another colleague suggested paraformaldehyde, but that is really inconvenient (preparing this every morning!?) Any advice on a good commercial fixative for this purpose, that doesn't destroy vessel antigens? Thanks. Min-Han This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient(s) please contact the sender by reply email and destroy all copies of the original message. Thank you. From ttroyer <@t> pcllab.com Wed Oct 27 12:30:35 2004 From: ttroyer <@t> pcllab.com (Travis Troyer) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] OLD CHEMICALS Message-ID: <001201c4bc4a$ab52f940$6601010a@Peterson.local> We have a cabinet full of older unused chemicals that we are no longer in need of. I was wondering if anyone would be interested in aquiring some of them or if anyone had a source I could use to dispose of them. If you are interested I could send you a list of the chemicals and stains we have. Thanks, Travis Troyer Peterson Clinical Lab From mward <@t> wfubmc.edu Wed Oct 27 12:34:53 2004 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] antibodies Message-ID: <61135F0455D33347B5AAE209B903A3040AB11EAA@EXCHVS2.medctr.ad.wfubmc.edu> I have been asked to look into two antibodies: Glut-1 and D2/40. Is anyone using them and if so could you recommend a vendor, etc. We have the capability of using the Ventana Nexes and the Dako autostainer. Any help you can give me would be greatly appreciated. Thanks! Martha Ward, MT (ASCP) QIHC Wake Forest University Baptist Medical Center From RCazares <@t> schosp.org Wed Oct 27 12:45:04 2004 From: RCazares <@t> schosp.org (Cazares, Ruth) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Thanks! Zinc formalin and bone marrows Message-ID: <913FAC2B773C19488E26AE6572180FA53AF951@exch01.schosp.org> Thanks for the info on bone marrow handling. For those of you who use zinc formalin for your bone marrows, my question is how long do you fix in this? And also what is the maximum time a core can stay in zinc formalin? We make up the bone marrow kits and if I replace the NBF with zinc formalin, sometimes the cores will sit in the fixative overnight (if done after we've gone home) and I'm wondering if this will damage the tissue. I have searched the archives, but I can't find a maximum time for tissues in ZF. Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. From ljb <@t> medicine.wisc.edu Wed Oct 27 13:47:45 2004 From: ljb <@t> medicine.wisc.edu (LaCinda Burchell) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Eosinophil marker, I have a great Ab Message-ID: Hi All, I absolutely understand and agree with John regarding the ability of even a simple H&E being able to stain Eos well. HOWEVER, I work in an area of research where eos and neuts are everything. My employers and supervisors insist that they must have IHC labeling also. So, I've had to find antibodies to do what a good H&E can do. If someone else is in the same position as me I suggest BD Pharmingen's mouse anti-human eosinophil Peroxidase. It stains beautifully for me at 1:100 without any HIER needed. What's more I'm able to use this antibody on Prefer ( Anatech ) fixed paraffin sections. When tested side-by-side the Prefer fixed sections where much nicer than the Formalin fixed sections! Dear UniPath IHC ; I'd be very happy to share more details with you. Including a couple other inexpensive special stains which are great too. Best of luck, Cindy B LaCinda Burchell, BA, AS, HT(ASCP) University of Wisconsin-Madison, Medical School Asthma and Allergy Research IHC Lab 600 Highland Ave. CSC K4/913 Madison, Wisconsin 53792 Phone: 608-262-3518 FAX: 608-263-3746 From ganglv16 <@t> hotmail.com Wed Oct 27 13:48:27 2004 From: ganglv16 <@t> hotmail.com (=?gb2312?B?wsAguNU=?=) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] C-Myc IHC Message-ID: Hello all, Anyone has experience on c-myc IHC? I just tried an Ab from Cell Signalling on mouse lung and it does not work quite well. Also I found the result of immunostaining on lung is not as good as GT tract. Anyone knows why? And how to improve it? Thanks a lot! Gary _________________________________________________________________ Ãâ·ÑÏÂÔØ [1]MSN Explorer Get 2 months FREE*. References 1. http://g.msn.com/8HMBCNCN/2746??PS=47575 From ajennings <@t> unmc.edu Wed Oct 27 13:50:10 2004 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Any fixative to reduce background in immunofluorescence? In-Reply-To: Message-ID: Clarification please are you using a secondary that is excited between 470-490? FITC? if so........does your background fluorescence appear to be more yellow than green? "paraformaldehyde" is not going to fix this particular problem is it confluent over the entire tissue? found in the blood cells? it may be as simple of a fix as changing your tag to something that excites 530-550 anita "Tan, MinHan" To Sent by: histonet-bounces@ cc lists.utsouthwest ern.edu Subject [Histonet] Any fixative to reduce background in immunofluorescence? 10/27/2004 12:14 PM Hi, I am preparing to do some immunofluorescence staining in kidney tissue with cd31. Some preliminary work in formalin fixed paraffin embedded tissue has yielded quite significant background fluorescence. A colleague suggested that I use frozen tissue, but the morphology is quite unacceptable. Another colleague suggested paraformaldehyde, but that is really inconvenient (preparing this every morning!?) Any advice on a good commercial fixative for this purpose, that doesn't destroy vessel antigens? Thanks. Min-Han This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient(s) please contact the sender by reply email and destroy all copies of the original message. Thank you. From pturner <@t> hsc.wvu.edu Wed Oct 27 14:54:47 2004 From: pturner <@t> hsc.wvu.edu (Pat Turner) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Looking for AFAP-110 antibody/vendor Message-ID: This antibody marks actin filament...I want to do IHC on mice From Ken.Matthews <@t> agresearch.co.nz Wed Oct 27 22:52:12 2004 From: Ken.Matthews <@t> agresearch.co.nz (Matthews, Ken) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Evans blue dye and myocardial infarction Message-ID: <4105FF527D8A1D4081C07B1147BF741503FFA5A6@rocket.agresearch.co.nz> I hope that someone out there may be able to shed light on our problem. We are studying means to reduce the tissue damage caused by heart attacks. Our studies suggest that, following the induction of a heart attack in our sheep model, some of the surviving tissue is compromised. We are attempting to delineate the areas of compromised tissue by post-mortem infusion of 0.15% Evans blue dye in 0.9% saline (we have also tried phosphate-buffered saline). The dye is infused into the coronary arteries immediately after death, followed by flushing with 0.9% saline. Our understanding of Evans blue is that it cannot cross intact cell membranes and therefore will only enter compromised muscle fibres (it has been used in this way to mark dystrophic muscle fibres in the mdx mouse). Can anybody tell me why the whole heart is staining blue during infusion, and remaining so following flushing. This happens even in control hearts which have not suffered a heart attack, and therefore have only normal muscle. I would be very grateful if anybody could come up with some suggestions. Thank you. Ken Matthews PhD Scientist Functional Muscle Genomics Group AgResearch Ruakura Private Bag 3123 East Street Hamilton New Zealand Tel. 64-7-838-5753 Fax. 64-7-838-5536 ======================================================================= Attention: The information contained in this message and/or attachments from AgResearch Limited is intended only for the persons or entities to which it is addressed and may contain confidential and/or privileged material. Any review, retransmission, dissemination or other use of, or taking of any action in reliance upon, this information by persons or entities other than the intended recipients is prohibited by AgResearch Limited. If you have received this message in error, please notify the sender immediately. ======================================================================= From jkiernan <@t> uwo.ca Thu Oct 28 00:24:01 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Evans blue dye and myocardial infarction References: <4105FF527D8A1D4081C07B1147BF741503FFA5A6@rocket.agresearch.co.nz> Message-ID: <41808271.98AFE9D9@uwo.ca> Evans blue is a dye with a large presence on the Internet and in the peer-reviewed scientific literature. The properties of the dye can be found in reference works such as the Merck index or Conn's Biological Stains. In your experiments (quoted below) the whole heart is stained by Evans blue because: (a) there is plenty of collagen in the heart, and collagen molecules just love to cling to long disazo dye molecules, and (b) the animal and its heart were dead before you flushed out the blood; trypan blue is excluded only by living cells. The dye enters dead cells. Trypan and Evns' blue are the oldest stains for cell viability. A new application of an old, cheap dye is always good news. The large dye anions are excluded from living cells, which are not abundant in the hearts of saline-perfused dead sheep. That's why your procedure provides all-blue hearts. The literature of vital/mortal staining with trypan blue (and the almost identical dye Evans blue) is enormous, even in the last 5 years. Have you tried a Google search? Both dyes date from the 1890s and entered the field of biological staining early in the 20th century. Your "compromised" heart muscle cells probably incorporate Evans blue because they are dead, along with all the other cardiac muscle cells that kept the animal alive until it's heart stopped while you were washing out the blood with saline. You cite no sources for the techniques that are not working as expected. Have you thoroughly evaluated the peer-reviewed literature before blindly following a list of instructions left by a former graduate student? The usual academic method is to start with the most recent technique from a paper in a peer-reviewed journal. If that doesn't work, you check in books and other papers. When well informed you try carefully controlled modifications of the older methods in your own lab. -- John Kiernan London, Canada. __________________________________________________ "Matthews, Ken" wrote: > > I hope that someone out there may be able to shed light on our problem. > We are studying means to reduce the tissue damage caused by heart > attacks. Our studies suggest that, following the induction of a heart > attack in our sheep model, some of the surviving tissue is compromised. > We are attempting to delineate the areas of compromised tissue by > post-mortem infusion of 0.15% Evans blue dye in 0.9% saline (we have > also tried phosphate-buffered saline). The dye is infused into the > coronary arteries immediately after death, followed by flushing with > 0.9% saline. Our understanding of Evans blue is that it cannot cross > intact cell membranes and therefore will only enter compromised muscle > fibres (it has been used in this way to mark dystrophic muscle fibres in > the mdx mouse). Can anybody tell me why the whole heart is staining blue > during infusion, and remaining so following flushing. This happens even > in control hearts which have not suffered a heart attack, and therefore > have only normal muscle. > I would be very grateful if anybody could come up with some suggestions. > Thank you. > > Ken Matthews PhD > Scientist > Functional Muscle Genomics Group > AgResearch Ruakura > Private Bag 3123 > East Street > Hamilton > New Zealand > > Tel. 64-7-838-5753 > Fax. 64-7-838-5536 > > ======================================================================= > Attention: The information contained in this message and/or attachments > from AgResearch Limited is intended only for the persons or entities > to which it is addressed and may contain confidential and/or privileged > material. Any review, retransmission, dissemination or other use of, or > taking of any action in reliance upon, this information by persons or > entities other than the intended recipients is prohibited by AgResearch > Limited. If you have received this message in error, please notify the > sender immediately. > ======================================================================= > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PWebster <@t> hei.org Thu Oct 28 02:33:36 2004 From: PWebster <@t> hei.org (Webster, Paul) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] polyester wax Message-ID: <4E8C1F1E4E8FA748B5487C50C91695F001280092@63-194-44-18.hei.org> Hi, Does anyone have experience with polyester wax as a substitute embedding medium? Is it possible to section decalcified bone and does it really offer better results for immunohistology? Regards, Paul Webster. Paul Webster, Ph.D. House Ear Institute 2100 W. 3rd St. Los Angeles CA 90057 From arvind <@t> nbrc.res.in Thu Oct 28 02:49:11 2004 From: arvind <@t> nbrc.res.in (Arvind Pundir) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] RE: Histonet Digest, Vol 11, Issue 36 Message-ID: <7DE0291A47C4BC4D9CDEF65D5017CEF0A6FA@mail.nbrc.res.in> can anyone tell me the protocol for autoradiography for localising the opioid receptors in brain of avians thanks in advance Arvind NBRC, INDIA From c.m.vanderloos <@t> amc.uva.nl Thu Oct 28 02:58:03 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] RE: human Ab on human tumor Message-ID: <49715a49ac81.49ac8149715a@amc.uva.nl> Dear Wen, I think your observation is completely right. When doing human-on-human staining the endogenous immunoglobulins in the tissue section will cause the background. In your case there will be no human immunoglobulins around and indeed this yields a background-free result. If you want to absolutely sure your result isn't fake, you may label your primary antibody directly. Most convenient is a Zenon kit (http://www.probes.com/products/zenon/zenon.html). The labeling procedure with biotin- or Alexa fluorochromes labeled Fab fragments is comparable with the Dako ARKit for mouse-on-mouse staining. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From wen eng Date Tue, 26 Oct 2004 12:00:56 -0700 (PDT) To histonet Subject [Histonet] human Ab on human tumor Hi histonetters, Currently I am doing IHC using human Ab on human tumor. I know it would cause "messy" background. But this tumor was developed by injection human tumor cell to nude mice. The results came out pretty good. I didn't use conjugated primary Ab, but just traditional method. I think it's because the tumor was developed in mice, the tissue would not contain endogenous human serum that causes the "messy background", although the tumor is human tumor. Am I right on this? Or my stain result just "fake"? Hope I said it clearly. Could any expert give me help? Thanks in advance! From c.m.vanderloos <@t> amc.uva.nl Thu Oct 28 03:14:42 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] RE: Eosinophil marker Message-ID: <418de8417dc4.417dc4418de8@amc.uva.nl> Dear Brianna, According to the datasheet of Monosan (www.monosan.com) antibody MON6008 (clone BMK-13) this antibody is staining FFPE sections after treating the tissue section with pepsin; not with HIER! Perhaps this pepsin treatment is worthwhile to test with your Chemicon antibody. Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From "UniPath IHC" Date Wed, 27 Oct 2004 10:34:50 -0600 To Subject RE: [Histonet] Eosinophil marker I guess I should have made myself clear, and I admit I forgot to sign my name before I sent off the email. I'm an IHC tech working for a private company. However, I'm not just trying to get "free advice on the internet". I have been a member of the Histonet community for four years and have enjoyed reading and contributing to this service. I would never use a response from Histonet as anything but advice or a hint in the right direction. As for who I work for, I have never been told that working for a private lab as opposed to a hospital excluded me from Histonet. I am working on a research project for a couple of doctors at the National Jewish in Denver. I am trying to doublestain eosinophils in skin biopsies by IHC and am having no luck with the MBP antibody (Chemicon) on routinely processed FFPE. It works fine for me on frozen sections, and even a little bit on tissue fixed in 4% PFA. I found a few emails in the archives regarding eosinophil markers, but most people asking what I need to know had never been replied to, at least not via the Histonet. I have considered special stains to mark eosinophils, but the doctors I am working with would like to use IHC. If I cannot make this antibody (MBP) work on FFPE then I need to find another eosinophil marker that does. Any help would be appreciated. Thanks for your time, Brianna Jackson, BS, QIHC UniPath, LLC Denver, CO 303-512-2220 bjackson@unipathllc.com From BMolinari <@t> heart.thi.tmc.edu Thu Oct 28 06:23:57 2004 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox Message-ID: Good morning, yes it is! Thank you Red Sox! I hope no one minds my posting but I came in to the lab this morning on cloud 9 and had to share ! Betsy Molinari HT (ASCP) Texas Heart Institute Houston (but born and raised in Boston :-)) TX 832-355-6524 From JWEEMS <@t> sjha.org Thu Oct 28 06:36:21 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox Message-ID: <83AACDB0810528418AA106F9AE9B7F7E50741B@sjhaexc02.sjha.org> I was thrilled myself and I haven't even been to Boston! I know you homefolks are! Enjoy the victory. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Molinari, Betsy Sent: Thursday, October 28, 2004 7:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Red Sox Good morning, yes it is! Thank you Red Sox! I hope no one minds my posting but I came in to the lab this morning on cloud 9 and had to share ! Betsy Molinari HT (ASCP) Texas Heart Institute Houston (but born and raised in Boston :-)) TX 832-355-6524 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From Jackie.O'Connor <@t> abbott.com Thu Oct 28 06:36:52 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox Message-ID: Thanks, Red Sox! Old curses die hard-but sweet! Wait until next year! (Cubs/ Red Sox game at Wrigley June 10, 2005) Die Hard Cubs Fan - Jackie O' "Molinari, Betsy" Sent by: histonet-bounces@lists.utsouthwestern.edu 10/28/2004 06:23 AM To: cc: Subject: [Histonet] Red Sox Good morning, yes it is! Thank you Red Sox! I hope no one minds my posting but I came in to the lab this morning on cloud 9 and had to share ! Betsy Molinari HT (ASCP) Texas Heart Institute Houston (but born and raised in Boston :-)) TX 832-355-6524 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mbecker <@t> pathlabinc.com Thu Oct 28 06:58:50 2004 From: mbecker <@t> pathlabinc.com (Michele Becker) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox In-Reply-To: Message-ID: New England could not be happier!! The Red Sox are World Champs!! Bring on the curse hearse!!! Michele NH -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jackie M O'Connor Sent: Thursday, October 28, 2004 6:37 AM To: Molinari, Betsy Cc: histonet@lists.utsouthwestern.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Red Sox Thanks, Red Sox! Old curses die hard-but sweet! Wait until next year! (Cubs/ Red Sox game at Wrigley June 10, 2005) Die Hard Cubs Fan - Jackie O' "Molinari, Betsy" Sent by: histonet-bounces@lists.utsouthwestern.edu 10/28/2004 06:23 AM To: cc: Subject: [Histonet] Red Sox Good morning, yes it is! Thank you Red Sox! I hope no one minds my posting but I came in to the lab this morning on cloud 9 and had to share ! Betsy Molinari HT (ASCP) Texas Heart Institute Houston (but born and raised in Boston :-)) TX 832-355-6524 _______________________________________________ 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 From nena.dimaano <@t> stryker.com Thu Oct 28 07:07:15 2004 From: nena.dimaano <@t> stryker.com (Dimaano, Nena) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox Message-ID: <20BE9059B6FC2D4B9E8FF2A9E2C4EEBD13F318@HOS2KEXCHCL.howost.strykercorp.com> I am so glad Rex Sox won....eventhough I am the only one celebrating here in my lab... I am so glad I was able to survive 2 weeks of pressure from the Yankee fans. Nena NJ (a shout away from the Yankee stadium) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Michele Becker Sent: Thursday, October 28, 2004 7:59 AM To: Jackie M O'Connor; Molinari, Betsy Cc: histonet@lists.utsouthwestern.edu; histonet-bounces@lists.utsouthwestern.edu Subject: RE: [Histonet] Red Sox New England could not be happier!! The Red Sox are World Champs!! Bring on the curse hearse!!! Michele NH -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jackie M O'Connor Sent: Thursday, October 28, 2004 6:37 AM To: Molinari, Betsy Cc: histonet@lists.utsouthwestern.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Red Sox Thanks, Red Sox! Old curses die hard-but sweet! Wait until next year! (Cubs/ Red Sox game at Wrigley June 10, 2005) Die Hard Cubs Fan - Jackie O' "Molinari, Betsy" Sent by: histonet-bounces@lists.utsouthwestern.edu 10/28/2004 06:23 AM To: cc: Subject: [Histonet] Red Sox Good morning, yes it is! Thank you Red Sox! I hope no one minds my posting but I came in to the lab this morning on cloud 9 and had to share ! Betsy Molinari HT (ASCP) Texas Heart Institute Houston (but born and raised in Boston :-)) TX 832-355-6524 _______________________________________________ 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 From funderwood <@t> mcohio.org Thu Oct 28 07:12:11 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:13 2005 Subject: [BULK] - [Histonet] Red Sox Message-ID: I'm not one to toot my own horn, but. I made my first trip to Boston 2 weekends ago, which included a tour of Fenway. The next thing you know the Sox win it all. Do you really think it's a coincidence? No thanks are necessary. Just remember the good karma of the Frederoo will trump the curse of the Bambino everytime. >>> "Molinari, Betsy" 10/28/04 07:23AM >>> Good morning, yes it is! Thank you Red Sox! I hope no one minds my posting but I came in to the lab this morning on cloud 9 and had to share ! Betsy Molinari HT (ASCP) Texas Heart Institute Houston (but born and raised in Boston :-)) TX 832-355-6524 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Thu Oct 28 07:24:46 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:13 2005 Subject: [BULK] - [Histonet] Red Sox Message-ID: Fred, have you ever been to Wrigley? My daughter works there - she'll be glad to give you a tour if you can guarantee the same results. "Fred Underwood" Sent by: histonet-bounces@lists.utsouthwestern.edu 10/28/2004 07:12 AM To: , cc: Subject: Re: [BULK] - [Histonet] Red Sox I'm not one to toot my own horn, but. I made my first trip to Boston 2 weekends ago, which included a tour of Fenway. The next thing you know the Sox win it all. Do you really think it's a coincidence? No thanks are necessary. Just remember the good karma of the Frederoo will trump the curse of the Bambino everytime. >>> "Molinari, Betsy" 10/28/04 07:23AM >>> Good morning, yes it is! Thank you Red Sox! I hope no one minds my posting but I came in to the lab this morning on cloud 9 and had to share ! Betsy Molinari HT (ASCP) Texas Heart Institute Houston (but born and raised in Boston :-)) TX 832-355-6524 _______________________________________________ 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 From BMolinari <@t> heart.thi.tmc.edu Thu Oct 28 07:27:55 2004 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:24:13 2005 Subject: [BULK] - [Histonet] Red Sox Message-ID: Yes, the Cubbies need to be next in line!!!!!!!!!!!! I can sympathize !!!!!!!!!!!!!!!!! -----Original Message----- From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] Sent: Thursday, October 28, 2004 7:25 AM To: Fred Underwood Cc: Molinari, Betsy; histonet@lists.utsouthwestern.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [BULK] - [Histonet] Red Sox Fred, have you ever been to Wrigley? My daughter works there - she'll be glad to give you a tour if you can guarantee the same results. "Fred Underwood" Sent by: histonet-bounces@lists.utsouthwestern.edu 10/28/2004 07:12 AM To: , cc: Subject: Re: [BULK] - [Histonet] Red Sox I'm not one to toot my own horn, but. I made my first trip to Boston 2 weekends ago, which included a tour of Fenway. The next thing you know the Sox win it all. Do you really think it's a coincidence? No thanks are necessary. Just remember the good karma of the Frederoo will trump the curse of the Bambino everytime. >>> "Molinari, Betsy" 10/28/04 07:23AM >>> Good morning, yes it is! Thank you Red Sox! I hope no one minds my posting but I came in to the lab this morning on cloud 9 and had to share ! Betsy Molinari HT (ASCP) Texas Heart Institute Houston (but born and raised in Boston :-)) TX 832-355-6524 _______________________________________________ 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 From BMolinari <@t> heart.thi.tmc.edu Thu Oct 28 07:41:55 2004 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:24:13 2005 Subject: [BULK] - [Histonet] Red Sox Message-ID: Did you mean White Sox or Cubs? For both it has been a long time. -----Original Message----- From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] Sent: Thursday, October 28, 2004 7:25 AM To: Fred Underwood Cc: Molinari, Betsy; histonet@lists.utsouthwestern.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [BULK] - [Histonet] Red Sox Fred, have you ever been to Wrigley? My daughter works there - she'll be glad to give you a tour if you can guarantee the same results. "Fred Underwood" Sent by: histonet-bounces@lists.utsouthwestern.edu 10/28/2004 07:12 AM To: , cc: Subject: Re: [BULK] - [Histonet] Red Sox I'm not one to toot my own horn, but. I made my first trip to Boston 2 weekends ago, which included a tour of Fenway. The next thing you know the Sox win it all. Do you really think it's a coincidence? No thanks are necessary. Just remember the good karma of the Frederoo will trump the curse of the Bambino everytime. >>> "Molinari, Betsy" 10/28/04 07:23AM >>> Good morning, yes it is! Thank you Red Sox! I hope no one minds my posting but I came in to the lab this morning on cloud 9 and had to share ! Betsy Molinari HT (ASCP) Texas Heart Institute Houston (but born and raised in Boston :-)) TX 832-355-6524 _______________________________________________ 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 From Andrew.Prior <@t> Smith-Nephew.com Thu Oct 28 08:22:14 2004 From: Andrew.Prior <@t> Smith-Nephew.com (Prior, Andrew) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Re: Exakt cutting system Message-ID: Dear Histonetters, Just wanted to sat thank you for all the replies to my query about the Exact system. Everyone seemed to have a positive view of it. Thank you to those people who have offered advice - I'll probably be in touch in January, once the system has arrived and we've had some training. If I get any useful info then I'll post in on the list for general reference. Andrew PS To the American contingent - Good Luck for the elections and don't forget to vote. (Who knows, you may get a President that was actually democratically elected this time! Not that that guarantees a decent leader - we still got Tony Blair!) PPS I hope I haven't opened a huge can of worms with my political views. No offence meant. Andrew Prior Histologist Smith & Nephew Research Centre York Science Park Heslington York Confidentiality. This electronic transmission is strictly confidential to Smith & Nephew and intended solely for the addressee. It may contain information which is covered by legal, professional or other privilege. If you are not the intended addressee, or someone authorised by the intended addressee to receive transmissions on behalf of the addressee, you must not retain, disclose in any form, copy or take any action in reliance on this transmission. If you have received this transmission in error, please notify the sender as soon as possible and destroy this message. From Luis.Chiriboga <@t> med.nyu.edu Thu Oct 28 07:48:23 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox In-Reply-To: Message-ID: I'm a die hard Yankee fan (Born in the Bronx)......& just want to pass along a congratulations to all the red sox fans. That was one of the best post season runs ever. I wouldn't of had it happen any other way...... -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Molinari, Betsy Sent: Thursday, October 28, 2004 7:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Red Sox Good morning, yes it is! Thank you Red Sox! I hope no one minds my posting but I came in to the lab this morning on cloud 9 and had to share ! Betsy Molinari HT (ASCP) Texas Heart Institute Houston (but born and raised in Boston :-)) TX 832-355-6524 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Thu Oct 28 08:24:55 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox In-Reply-To: Message-ID: Betsy, I used to skip school and go to Fenway. I hope it's not another 86 years before the Red Sox wins again. I never thought that I would see the Patriots and Red Sox win a championship in the same year. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Molinari, Betsy Sent: Thursday, October 28, 2004 6:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Red Sox Good morning, yes it is! Thank you Red Sox! I hope no one minds my posting but I came in to the lab this morning on cloud 9 and had to share ! Betsy Molinari HT (ASCP) Texas Heart Institute Houston (but born and raised in Boston :-)) TX 832-355-6524 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Thu Oct 28 08:26:26 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:13 2005 Subject: [BULK] - [Histonet] Red Sox In-Reply-To: Message-ID: Fred, you need to go back to Boston next October so we can celebrate again!!!!!! Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Fred Underwood Sent: Thursday, October 28, 2004 7:12 AM To: BMolinari@heart.thi.tmc.edu; histonet@lists.utsouthwestern.edu Subject: Re: [BULK] - [Histonet] Red Sox I'm not one to toot my own horn, but. I made my first trip to Boston 2 weekends ago, which included a tour of Fenway. The next thing you know the Sox win it all. Do you really think it's a coincidence? No thanks are necessary. Just remember the good karma of the Frederoo will trump the curse of the Bambino everytime. >>> "Molinari, Betsy" 10/28/04 07:23AM >>> Good morning, yes it is! Thank you Red Sox! I hope no one minds my posting but I came in to the lab this morning on cloud 9 and had to share ! Betsy Molinari HT (ASCP) Texas Heart Institute Houston (but born and raised in Boston :-)) TX 832-355-6524 _______________________________________________ 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 From JNocito <@t> Pathreflab.com Thu Oct 28 08:27:12 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:13 2005 Subject: [BULK] - [Histonet] Red Sox In-Reply-To: Message-ID: Jackie, I'm sorry, Fred will be tied up next year (even if I have to do it myself) Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jackie M O'Connor Sent: Thursday, October 28, 2004 7:25 AM To: Fred Underwood Cc: histonet@lists.utsouthwestern.edu; BMolinari@heart.thi.tmc.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [BULK] - [Histonet] Red Sox Fred, have you ever been to Wrigley? My daughter works there - she'll be glad to give you a tour if you can guarantee the same results. "Fred Underwood" Sent by: histonet-bounces@lists.utsouthwestern.edu 10/28/2004 07:12 AM To: , cc: Subject: Re: [BULK] - [Histonet] Red Sox I'm not one to toot my own horn, but. I made my first trip to Boston 2 weekends ago, which included a tour of Fenway. The next thing you know the Sox win it all. Do you really think it's a coincidence? No thanks are necessary. Just remember the good karma of the Frederoo will trump the curse of the Bambino everytime. >>> "Molinari, Betsy" 10/28/04 07:23AM >>> Good morning, yes it is! Thank you Red Sox! I hope no one minds my posting but I came in to the lab this morning on cloud 9 and had to share ! Betsy Molinari HT (ASCP) Texas Heart Institute Houston (but born and raised in Boston :-)) TX 832-355-6524 _______________________________________________ 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 From c.m.vanderloos <@t> amc.uva.nl Thu Oct 28 08:46:12 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] RE: antibodies Message-ID: <4648fb465873.4658734648fb@amc.uva.nl> Martha, We have good results with: Glut-1 (rabbit) from Dako (A3536) D2/40 from Signet Labs (www.signetlabs.com) Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Martha Ward Date Wed, 27 Oct 2004 13:34:53 -0400 To histonet@lists.utsouthwestern.edu Subject [Histonet] antibodies I have been asked to look into two antibodies: Glut-1 and D2/40. Is anyone using them and if so could you recommend a vendor, etc. We have the capability of using the Ventana Nexes and the Dako autostainer. Any help you can give me would be greatly appreciated. Thanks! Martha Ward, MT (ASCP) QIHC Wake Forest University Baptist Medical Center From herme013 <@t> umn.edu Thu Oct 28 08:46:50 2004 From: herme013 <@t> umn.edu (Yves Heremans) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] (no subject) Message-ID: <200410281346.i9SDko2S013913@challenge.software.umn.edu> Dear Histonetters, Is anybody aware of good species-specific antibodies against rat and mouse MHC-antigens ? We are planning to co-culture rat and mouse cells and are looking for ways to distinguish both by staining. Yves Yves Heremans, Ph.D. University of Minnesota Stem Cell Institute Tel 612-625-0964 Fax 612-624-2436 Address for US Postal Mail: University of Minnesota MMC 716 420 Delaware Street SE Minneapolis, MN 55455 Address for COURIER DELIVERY: Suite 14-285 Moos Tower 515 Delaware Street SE   Yves Heremans, Ph.D. University of Minnesota Stem Cell Institute Tel 612-625-0964 Fax 612-624-2436 Address for US Postal Mail: University of Minnesota MMC 716 420 Delaware Street SE Minneapolis, MN 55455 Address for COURIER DELIVERY: Suite 14-285 Moos Tower 515 Delaware Street SE From funderwood <@t> mcohio.org Thu Oct 28 08:54:09 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:13 2005 Subject: [BULK] - [Histonet] Red Sox Message-ID: I'd be happy to help out both cities. But if you've seen the performances of ALL Ohio sports teams, I'll probably be drained by next spring. >>> "Joe Nocito" 10/28/04 09:27AM >>> Jackie, I'm sorry, Fred will be tied up next year (even if I have to do it myself) Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jackie M O'Connor Sent: Thursday, October 28, 2004 7:25 AM To: Fred Underwood Cc: histonet@lists.utsouthwestern.edu; BMolinari@heart.thi.tmc.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [BULK] - [Histonet] Red Sox Fred, have you ever been to Wrigley? My daughter works there - she'll be glad to give you a tour if you can guarantee the same results. "Fred Underwood" Sent by: histonet-bounces@lists.utsouthwestern.edu 10/28/2004 07:12 AM To: , cc: Subject: Re: [BULK] - [Histonet] Red Sox I'm not one to toot my own horn, but. I made my first trip to Boston 2 weekends ago, which included a tour of Fenway. The next thing you know the Sox win it all. Do you really think it's a coincidence? No thanks are necessary. Just remember the good karma of the Frederoo will trump the curse of the Bambino everytime. >>> "Molinari, Betsy" 10/28/04 07:23AM >>> Good morning, yes it is! Thank you Red Sox! I hope no one minds my posting but I came in to the lab this morning on cloud 9 and had to share ! Betsy Molinari HT (ASCP) Texas Heart Institute Houston (but born and raised in Boston :-)) TX 832-355-6524 _______________________________________________ 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 From ljb <@t> medicine.wisc.edu Thu Oct 28 09:03:14 2004 From: ljb <@t> medicine.wisc.edu (LaCinda Burchell) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] The Ab and a couple of special stains Message-ID: Good Morning Brianna, The Catalog number for the BD Pharmingen Ab is #554252. Again, I use it at a dilution of 1:100 with streptavidin HRP or ALP reagents from Biocare Medical. I incubate the primary for 1 hour at room temp and, incubate the secondary and tertiary for 15 mins each. I'm also attaching a couple of other special stains that I've found over the years for eosinophils. All my Best!! Cindy B LaCinda Burchell, BA, AS, HT(ASCP) University of Wisconsin-Madison, Medical School Asthma and Allergy Research IHC Lab 600 Highland Ave. CSC K4/913 Madison, Wisconsin 53792 Phone: 608-262-3518 FAX: 608-263-3746 From funderwood <@t> mcohio.org Thu Oct 28 09:07:04 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:13 2005 Subject: [BULK] - [Histonet] OLD CHEMICALS Message-ID: Check with your county/local waste processing center. In addition to old paint and oil, you'll probably be suprised what else they will dispose of for you. Fred >>> "Travis Troyer" 10/27/04 01:30PM >>> We have a cabinet full of older unused chemicals that we are no longer in need of. I was wondering if anyone would be interested in aquiring some of them or if anyone had a source I could use to dispose of them. If you are interested I could send you a list of the chemicals and stains we have. Thanks, Travis Troyer Peterson Clinical Lab _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Lynne.Bell <@t> hitchcock.org Thu Oct 28 09:19:33 2004 From: Lynne.Bell <@t> hitchcock.org (Bell, Lynne) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox Message-ID: I don't mind one bit, Betsy. I'm a lifelong (50 years) Red Sox Fan!! I've watched or been at every game this year.............I'm working on about 3 1/2 hours of sleep today, but it is well worth it! GO RED SOX!! Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 From ian.montgomery <@t> bio.gla.ac.uk Thu Oct 28 09:35:11 2004 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox. Message-ID: <6.1.2.0.2.20041028153102.02d3b8f0@udcf.gla.ac.uk> Who are the Red Sox and do they play the children's game rounders? Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07625 702883 e-mail: ian.montgomery@bio.gla.ac.uk From vitha <@t> mic.tamu.edu Thu Oct 28 09:46:23 2004 From: vitha <@t> mic.tamu.edu (Stanislav Vitha) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Re: Polyester wax In-Reply-To: <200410281328.i9SDSkAD030184@smtp-relay.tamu.edu> References: <200410281328.i9SDSkAD030184@smtp-relay.tamu.edu> Message-ID: <6.1.2.0.0.20041028091016.03325880@mic.tamu.edu> Hi, I have used the polyester wax (Steedman's wax) a lot for immunofluorescence in plants (with antibodies for actin, tubulin, FtsZ, GFP, c-myc, and other). I remember there were some old papers where the wax was used for decalcified bones. Unfortunately these reprints were left behind during my last move, but a quick search found a couple papers that may be relevant: 1. Sage, M., Polyethylene-Glycol Distearate 600 with 10 Percent 1-Hexadecanol - Superior Embedding Wax for Warm Climates. Stain Technology, 1972. 47(6): p. 313-&. 2. Goodwin, J.R., Polyester Wax Processing of Tissues - Some New Techniques and Detailed Procedures. American Journal of Medical Technology, 1974. 40(2): p. 45-49. 3. Josephse.K, Technique for Isolating Enamel Organ of Rat Incisor for Histologic Studies. Scandinavian Journal of Dental Research, 1974. 82(3): p. 229-238. 4. Moores, B.D., Thin Histological Sections from Tissues Impregnated with Digol Distearate. Medical Laboratory Sciences, 1976. 33(1): p. 73-77. 5. Kusakabe, M., Sakakura, T., Nishizuka, Y., Sano, M., and Matsukage, A., Polyester Wax Embedding and Sectioning Technique for Immunohistochemistry. Stain Technology, 1984. 59(3): p. 127-132. 6. Roholl, P.J.M., Dullens, H.F.J., Kleijne, J., Dubbink, E.J., and Denotter, W., Acid Ethanol Fixation and Polyester Wax Embedding Combines Preservation of Antigenic Determinants with Good Morphology and Enables Simultaneous Bromodeoxyuridine (Brdu) Labeling. Biotechnic & Histochemistry, 1991. 66(2): p. 55-62. 7. Richardson, L.L. and Dym, M., Improved Adhesiveness of Polyester Wax Sections for Immunocytochemistry. Biotechniques, 1994. 17(5): p. 846-848. As far as handling and sectioning, I was quite happy with the sectioning properties; I was cutting sections between 5 and 15 microns. There are few tricks that are useful to make the sectioning work well; see the book chapter: Vitha, S., Balu?ka, F., Jasik, J., Volkmann, D., and Barlow, P., Steedman's wax for F-actin visualization, in Actin: a Dynamic Framework for Multiple Plant Cell Functions, C.J. Staiger, F. Balu?ka, D. Volkmann, and P. Barlow, Editors. 2000, Kluwer: Dordrecht, The Netherlands. p. 619-636. You can download the PDF from http://www.izmb.de/volkmann/pdfs/ (Book-Steedman's_wax.pdf) The disadvantage of the above method is that the ribbons sometimes do not expand 100%, or al least not all sections; for what I was doing it was not critical, I was more interested in cellular and subcellular immunostaining rather that looking at tissue and whole-organ morphology. Also see my previous post to this forum (10-21-2004) Stan Vitha >Message: 11 >Date: Thu, 28 Oct 2004 00:33:36 -0700 >From: "Webster, Paul" >Subject: [Histonet] polyester wax >To: >Message-ID: > <4E8C1F1E4E8FA748B5487C50C91695F001280092@63-194-44-18.hei.org> >Content-Type: text/plain; charset="Windows-1252" > >Hi, > >Does anyone have experience with polyester wax as a substitute embedding >medium? Is it possible to section decalcified bone and does it really >offer better results for immunohistology? > >Regards, > >Paul Webster. > > >Paul Webster, Ph.D. >House Ear Institute >2100 W. 3rd St. >Los Angeles >CA 90057 > Dr. Stanislav Vitha vitha@mic.tamu.edu Microscopy and Imaging Center Texas A&M University BSBW 119 College Station, TX 77843-2257 tel: 979-845-1129 (main desk) tel: 979-845-1607 (direct link) fax: 979-847-8933 From jengirl1014 <@t> yahoo.com Thu Oct 28 09:59:46 2004 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox Message-ID: <20041028145946.74752.qmail@web60604.mail.yahoo.com> Just wanted to congratulate Boston for sticking by their team all these years. As a Baltimorean, I prayed every night that God would talk some sense into the Great Bambino and let them win. I'm glad to see it worked. Take the day off Massachusettes!!! You've earned it!!! Jennifer K. Sipes, RALAT Sr. Laboratory Technician Johns Hopkins University Ross 929 720 Rutland Avenue Baltimore, MD 21205 phone: 410-614-0131 cell: 443-413-0853 e-mail: jengirl1014@yahoo.com --------------------------------- Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. From pam <@t> ategra.com Thu Oct 28 10:23:00 2004 From: pam <@t> ategra.com (Pam Barker (extension 234)) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Histo Tech and Pathology Assistant needed in the Dallas/Ft. Worth area Message-ID: Histonetters - I am presently on a search for one of my best clients in the Dallas/Ft. Worth area who is seeking to hire a Histo tech and a Pathology Assistant. These are full time (40 hours per week) permanent positions. My client offers competitive salaries and excellent benefits. Are you interested ? Also, if you have friends/peers who might be interested, if you could pass my query & name on to them I'd be much obliged. If interested, please call me at 800-466-9919 x234. Thank You !! Pam - 800 466 9919 ext 234 --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing Learn More About Ategra: http://www.ategra.com Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd Winter Park, FL 32792-6721 800-466-9919 ext 234 EMAIL: pam@ategra.com WEBSITE: http://www.ategra.com -------------------------------------------------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. -------------------------------------------------------------------------------------------------- From ian.montgomery <@t> bio.gla.ac.uk Thu Oct 28 10:35:31 2004 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:24:13 2005 Subject: Fwd: RE: [Histonet] Red Sox. Message-ID: <6.1.2.0.2.20041028162403.02d32a30@udcf.gla.ac.uk> >What are children's game rounders? > Almost identical to baseball but strictly for children. I've always been surprised that the pioneers of the Wild West would end up playing a game for children. Still, when big bucks are involved men being the weaker sex would do anything. >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ian >Montgomery >Sent: Thursday, October 28, 2004 9:35 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Red Sox. > > Who are the Red Sox and do they play the children's game >rounders? > >Dr. Ian Montgomery, >Histotechnology, >Graham Kerr Building, >Institute of Biomedical & Life Sciences, >University of Glasgow, >Glasgow, >G12 8QQ. >Tel: 0141 339 8855 >Office: 4652 >Lab: 6644. >Pager: 07625 702883 >e-mail: ian.montgomery@bio.gla.ac.uk > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07625 702883 e-mail: ian.montgomery@bio.gla.ac.uk From pruegg <@t> ihctech.net Thu Oct 28 10:55:44 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] RE: Eosinophil marker In-Reply-To: <418de8417dc4.417dc4418de8@amc.uva.nl> Message-ID: <001f01c4bd06$965e4370$83020a0a@IHCTech> Eosin is the best marker for eosinophils I know of. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of C.M. van der Loos Sent: Thursday, October 28, 2004 1:15 AM To: histonet@lists.utsouthwestern.edu Cc: ihc@unipathllc.com Subject: [Histonet] RE: Eosinophil marker Dear Brianna, According to the datasheet of Monosan (www.monosan.com) antibody MON6008 (clone BMK-13) this antibody is staining FFPE sections after treating the tissue section with pepsin; not with HIER! Perhaps this pepsin treatment is worthwhile to test with your Chemicon antibody. Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From "UniPath IHC" Date Wed, 27 Oct 2004 10:34:50 -0600 To Subject RE: [Histonet] Eosinophil marker I guess I should have made myself clear, and I admit I forgot to sign my name before I sent off the email. I'm an IHC tech working for a private company. However, I'm not just trying to get "free advice on the internet". I have been a member of the Histonet community for four years and have enjoyed reading and contributing to this service. I would never use a response from Histonet as anything but advice or a hint in the right direction. As for who I work for, I have never been told that working for a private lab as opposed to a hospital excluded me from Histonet. I am working on a research project for a couple of doctors at the National Jewish in Denver. I am trying to doublestain eosinophils in skin biopsies by IHC and am having no luck with the MBP antibody (Chemicon) on routinely processed FFPE. It works fine for me on frozen sections, and even a little bit on tissue fixed in 4% PFA. I found a few emails in the archives regarding eosinophil markers, but most people asking what I need to know had never been replied to, at least not via the Histonet. I have considered special stains to mark eosinophils, but the doctors I am working with would like to use IHC. If I cannot make this antibody (MBP) work on FFPE then I need to find another eosinophil marker that does. Any help would be appreciated. Thanks for your time, Brianna Jackson, BS, QIHC UniPath, LLC Denver, CO 303-512-2220 bjackson@unipathllc.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From amosbrooks <@t> earthlink.net Thu Oct 28 11:08:39 2004 From: amosbrooks <@t> earthlink.net (amosbrooks@earthlink.net) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Re: Histonet Digest, Vol 11, Issue 37 Message-ID: <25283750.1098979719841.JavaMail.root@statler.psp.pas.earthlink.net> Martha, For Glut-1 try Dako A3536 at 1:100 using Envision (or some such secondary). It will require epitope retrieval. As for the other, we've not used it. Good luck, Amos Message: 3 Date: Wed, 27 Oct 2004 13:34:53 -0400 From: "Martha Ward" Subject: [Histonet] antibodies To: Message-ID: <61135F0455D33347B5AAE209B903A3040AB11EAA@EXCHVS2.medctr.ad.wfubmc.edu> Content-Type: text/plain; charset="US-ASCII" I have been asked to look into two antibodies: Glut-1 and D2/40. Is anyone using them and if so could you recommend a vendor, etc. We have the capability of using the Ventana Nexes and the Dako autostainer. Any help you can give me would be greatly appreciated. Thanks! Martha Ward, MT (ASCP) QIHC Wake Forest University Baptist Medical Center From amosbrooks <@t> earthlink.net Thu Oct 28 11:20:37 2004 From: amosbrooks <@t> earthlink.net (amosbrooks@earthlink.net) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox Message-ID: <5772910.1098980437494.JavaMail.root@statler.psp.pas.earthlink.net> Hi, Don't say ALL of New England is happy. Here's one Connecticut Yankees fan still reeling from four consecutive losses here. I can only say the win must have had something to do with the lunar eclipse last night. Personally given the signs: war, earthquakes in Japan, an eclipse and the Red Sox winning a World Series, I'm starting work on an ark because the end must be near! Congradulations Red Sox fans your team played exceptionally and deserve the pe... penn ... pennant ** there I said it** ick I'm feeling woozy! Amos Brooks From TJasper <@t> smdc.org Thu Oct 28 11:25:33 2004 From: TJasper <@t> smdc.org (Jasper, Thomas G.) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox. Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA02E44FF0@harrier> Hey Ian, You've obviously never taken a "high hard one to the noggin'". I only say that because as a child I did playing Little League Baseball. After that I asked my mother for a tennis racket. I do however admire the skill of Major League Baseball players. My congratulations to the Red Sox for lifting the curse of the Bambino. Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: Ian Montgomery [mailto:ian.montgomery@bio.gla.ac.uk] Sent: Thursday, October 28, 2004 10:36 AM To: histonet@lists.utsouthwestern.edu Subject: Fwd: RE: [Histonet] Red Sox. >What are children's game rounders? > Almost identical to baseball but strictly for children. I've always been surprised that the pioneers of the Wild West would end up playing a game for children. Still, when big bucks are involved men being the weaker sex would do anything. >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ian >Montgomery >Sent: Thursday, October 28, 2004 9:35 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Red Sox. > > Who are the Red Sox and do they play the children's game >rounders? > >Dr. Ian Montgomery, >Histotechnology, >Graham Kerr Building, >Institute of Biomedical & Life Sciences, >University of Glasgow, >Glasgow, >G12 8QQ. >Tel: 0141 339 8855 >Office: 4652 >Lab: 6644. >Pager: 07625 702883 >e-mail: ian.montgomery@bio.gla.ac.uk > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07625 702883 e-mail: ian.montgomery@bio.gla.ac.uk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From algranth <@t> u.arizona.edu Thu Oct 28 11:26:51 2004 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:24:13 2005 Subject: Fwd: RE: [Histonet] Red Sox. In-Reply-To: <6.1.2.0.2.20041028162403.02d32a30@udcf.gla.ac.uk> Message-ID: <4.3.2.7.2.20041028092159.00c8b490@algranth.inbox.email.arizona.edu> The big bucks play an important part. I just read where Curt Schilling negotiated for an extra $15 MILLION in his contract. As a histotech I can't imagine AN EXTRA $15 Million! Still...Congrats to the Red Sox and their fans all over the world. When the D'backs won the World Series they brought the trophy here and I got to put my fingerprints on it - even that was exciting! Now back to work... Andi At 04:35 PM 10/28/2004 +0100, Ian Montgomery wrote: >>What are children's game rounders? > > Almost identical to baseball but strictly for children. I've > always been surprised that the pioneers of the Wild West would end up > playing a game for children. Still, when big bucks are involved men being > the weaker sex would do anything. > > > > > >>-----Original Message----- >>From: histonet-bounces@lists.utsouthwestern.edu >>[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ian >>Montgomery >>Sent: Thursday, October 28, 2004 9:35 AM >>To: histonet@lists.utsouthwestern.edu >>Subject: [Histonet] Red Sox. >> >> Who are the Red Sox and do they play the children's game >>rounders? >> >>Dr. Ian Montgomery, >>Histotechnology, >>Graham Kerr Building, >>Institute of Biomedical & Life Sciences, >>University of Glasgow, >>Glasgow, >>G12 8QQ. >>Tel: 0141 339 8855 >>Office: 4652 >>Lab: 6644. >>Pager: 07625 702883 >>e-mail: ian.montgomery@bio.gla.ac.uk >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Dr. Ian Montgomery, >Histotechnology, >Graham Kerr Building, >Institute of Biomedical & Life Sciences, >University of Glasgow, >Glasgow, >G12 8QQ. >Tel: 0141 339 8855 >Office: 4652 >Lab: 6644. >Pager: 07625 702883 >e-mail: ian.montgomery@bio.gla.ac.uk > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From la.sebree <@t> hosp.wisc.edu Thu Oct 28 11:32:39 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Legionella Message-ID: Just wondering if there are any reference labs out there that immunostain for Legionella disease. Thanks, Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 From funderwood <@t> mcohio.org Thu Oct 28 11:38:25 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:13 2005 Subject: [BULK] - Re: Fwd: RE: [Histonet] Red Sox. Message-ID: 15 million dollars!!! Wow. In the histotech world, that's like getting a bonus box of disposable blades. >>> Andrea Grantham 10/28/04 12:26PM >>> The big bucks play an important part. I just read where Curt Schilling negotiated for an extra $15 MILLION in his contract. As a histotech I can't imagine AN EXTRA $15 Million! Still...Congrats to the Red Sox and their fans all over the world. When the D'backs won the World Series they brought the trophy here and I got to put my fingerprints on it - even that was exciting! Now back to work... Andi At 04:35 PM 10/28/2004 +0100, Ian Montgomery wrote: >>What are children's game rounders? > > Almost identical to baseball but strictly for children. I've > always been surprised that the pioneers of the Wild West would end up > playing a game for children. Still, when big bucks are involved men being > the weaker sex would do anything. > > > > > >>-----Original Message----- >>From: histonet-bounces@lists.utsouthwestern.edu >>[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ian >>Montgomery >>Sent: Thursday, October 28, 2004 9:35 AM >>To: histonet@lists.utsouthwestern.edu >>Subject: [Histonet] Red Sox. >> >> Who are the Red Sox and do they play the children's game >>rounders? >> >>Dr. Ian Montgomery, >>Histotechnology, >>Graham Kerr Building, >>Institute of Biomedical & Life Sciences, >>University of Glasgow, >>Glasgow, >>G12 8QQ. >>Tel: 0141 339 8855 >>Office: 4652 >>Lab: 6644. >>Pager: 07625 702883 >>e-mail: ian.montgomery@bio.gla.ac.uk >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Dr. Ian Montgomery, >Histotechnology, >Graham Kerr Building, >Institute of Biomedical & Life Sciences, >University of Glasgow, >Glasgow, >G12 8QQ. >Tel: 0141 339 8855 >Office: 4652 >Lab: 6644. >Pager: 07625 702883 >e-mail: ian.montgomery@bio.gla.ac.uk > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Thu Oct 28 11:47:32 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Legionella Message-ID: We do IHC for legionella here at Hartford Hospital. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Sebree Linda A." 10/28/04 12:32PM >>> Just wondering if there are any reference labs out there that immunostain for Legionella disease. Thanks, Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Lynne.Bell <@t> hitchcock.org Thu Oct 28 11:47:47 2004 From: Lynne.Bell <@t> hitchcock.org (Bell, Lynne) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox Message-ID: Amos, I know how hard it is to congratulate the "WORLD CHAMPIONS" - I'm overjoyed to be on the receiving end of the congratulations this year. Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 From akbitting <@t> geisinger.edu Thu Oct 28 12:31:22 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox Message-ID: Does anyone else out there think it has more to do with selling tickets, advertising time, etc. Don't you think "Baseball" knew that the Red Sox going to the W.S. would draw more of an audience than the Yankees winning again! It's all about the Benjamins! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 >>> 10/28/04 12:20PM >>> Hi, Don't say ALL of New England is happy. Here's one Connecticut Yankees fan still reeling from four consecutive losses here. I can only say the win must have had something to do with the lunar eclipse last night. Personally given the signs: war, earthquakes in Japan, an eclipse and the Red Sox winning a World Series, I'm starting work on an ark because the end must be near! Congradulations Red Sox fans your team played exceptionally and deserve the pe... penn ... pennant ** there I said it** ick I'm feeling woozy! Amos Brooks _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From pstefanova <@t> sten.sunnybrook.utoronto.ca Thu Oct 28 13:09:26 2004 From: pstefanova <@t> sten.sunnybrook.utoronto.ca (Petia P Stefanova) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] axolotl embryos References: Message-ID: <00a301c4bd19$43253340$9d194c8e@WS21203> Hi, Does anyone have experience with axolotl embryos? Is it possible before processing to stain whole embryo so I can get a 3D image and then continue with processing? If anyone has done this and has a protocol I would very much appreciated it. Thanks to all! PStefanova From JNocito <@t> Pathreflab.com Thu Oct 28 13:25:35 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox In-Reply-To: Message-ID: Angela, I too thought that way until the 4 game sweep. If it was money, this series would have gone 7 games. Just think about the lost revenue from TV and tickets sales. Truthfully, I haven't fallowed baseball since the last time they went on strike. I can't empathize why someone would strike for $30 mil instead of $25 mil. As a histo tech, I'll be lucking if I see $2 mil over my entire career. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Angela Bitting Sent: Thursday, October 28, 2004 12:31 PM To: amosbrooks@earthlink.net; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Red Sox Does anyone else out there think it has more to do with selling tickets, advertising time, etc. Don't you think "Baseball" knew that the Red Sox going to the W.S. would draw more of an audience than the Yankees winning again! It's all about the Benjamins! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 >>> 10/28/04 12:20PM >>> Hi, Don't say ALL of New England is happy. Here's one Connecticut Yankees fan still reeling from four consecutive losses here. I can only say the win must have had something to do with the lunar eclipse last night. Personally given the signs: war, earthquakes in Japan, an eclipse and the Red Sox winning a World Series, I'm starting work on an ark because the end must be near! Congradulations Red Sox fans your team played exceptionally and deserve the pe... penn ... pennant ** there I said it** ick I'm feeling woozy! Amos Brooks _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From histo007 <@t> hotmail.com Thu Oct 28 13:25:17 2004 From: histo007 <@t> hotmail.com (Jim Ball) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] A note to all the Boston elite Message-ID: As in histology the ability of being consistent is valued more highly than being lucky. I understand this is the first time Boston has won a world Series since WW-1, which brings me back to an old saying back home. "Even a blind Squirrel will find a nut from time to time". Oh by the way how many Times have the Yankees been to the big dance since Lets say the Vietnam era. I would'nt want any body to have to get out a calculator and start adding up the numbers from WW-1. Yes I am a Rebel at heart and a Yankee fan by choice. Cong "rats" Boston we'll see you next year, and hopefully it'll be the Yankees and Chicago in the Series. Now thats a real underdog, even I would have a hard time deciding who should win that one. _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! hthttp://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From BlazekL <@t> childrensdayton.org Thu Oct 28 13:26:18 2004 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox Message-ID: Shame on you for degrading the game of baseball! To imply that "baseball" is fixed must be some sort of crime! I think the same number of people would have watched if it had been the Yankees playing but not with the same kind of thrill. It's wonderful to see the underdogs come out on top once in awhile. And now the Bambino can rest. Linda >>> "Angela Bitting" 10/28/2004 1:31:22 PM >>> Does anyone else out there think it has more to do with selling tickets, advertising time, etc. Don't you think "Baseball" knew that the Red Sox going to the W.S. would draw more of an audience than the Yankees winning again! It's all about the Benjamins! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 >>> 10/28/04 12:20PM >>> Hi, Don't say ALL of New England is happy. Here's one Connecticut Yankees fan still reeling from four consecutive losses here. I can only say the win must have had something to do with the lunar eclipse last night. Personally given the signs: war, earthquakes in Japan, an eclipse and the Red Sox winning a World Series, I'm starting work on an ark because the end must be near! Congradulations Red Sox fans your team played exceptionally and deserve the pe... penn ... pennant ** there I said it** ick I'm feeling woozy! Amos Brooks _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From KTennako <@t> odu.edu Thu Oct 28 13:45:44 2004 From: KTennako <@t> odu.edu (Kushan U Tennakoon) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] wax embedding of plant tissues with latex Message-ID: Dear Histonetters, We have encountered problems when trying to embed Euphorbia root tissues (that have a lot of latex) in paraffin wax. Can you please advice me / give some fine points to ensure the optimal penetration of paraffin wax into the plant tissues that have a lot of milky latex. Sincerely, Kushan Tennakoon ktennako@odu.edu From vitha <@t> mic.tamu.edu Thu Oct 28 13:56:18 2004 From: vitha <@t> mic.tamu.edu (Stanislav Vitha) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Re: microscope eyepiece adaptor Message-ID: <6.1.2.0.0.20041028130139.0332fcc0@mic.tamu.edu> I have used the LensPlus adapter (http://www.lensadapter.com/products.htm) for our Coolpix 995 - it connects to your camera's 28mm filter thread and clamps around an eyepiece - in my case it was for an old Olympus BH2 microscope - since in older scopes the optical aberrations of the objective are often compensated by the oculars (or the projection photopiece for film photography), the "universal" third party optical adaptor element may not work that well. Therefore I used the Olympus eyepiece. However, on new scopes the objectives are well corrected. For the Leica DMRA I used the Optem (http://www.thales-optem.com/) adapter, which had a 28mm thread. I also tested a Nikon adapter, but it had a serious problem - the front element of the adapter was recessed too much (several mm), and thus to avoid vignetting you had to zoom in with the camera quite a bit, losing thus the field of view coverage. Another source to consider is the Zarf Enterprises http://zarfenterprises.com/lens%20adapter%20series.html In our setup, we connected a cheap 13-in TV to the video output from the camera to aid focusing. We initially had a remote release from Nikon (~$90), but in the end I set up an old linux computer connected via the Coolpix serial cable to the camera and wrote a simple shell script to control the free "photopc: software) to allow users take pictures with a keystroke, and, importantly, set the zoom of the camera to a precise numerical value - this way you can directly compare image scale between different imaging sessions and do not have to take a picture of the stage micrometer every time. Also check the newsgroup on yahoo, there are links and discussion on software and adapters http://groups.yahoo.com/group/CoolpixPhotoMicMac/ More info on photopc and the optional graphical user interface for linux and windows, see http://www.math.ualberta.ca/imaging/ Stan Vitha Dr. Stanislav Vitha vitha@mic.tamu.edu Microscopy and Imaging Center Texas A&M University BSBW 119 College Station, TX 77843-2257 tel: 979-845-1129 (main desk) tel: 979-845-1607 (direct link) fax: 979-847-8933 >-----Original Message----- >From: Kushan U Tennakoon [mailto:KTennako at odu.edu] >Sent: Tuesday, October 12, 2004 12:39 PM >To: histonet at >lists.utsouthwestern.edu >Subject: [Histonet] microscope eyepiece adaptor > >We need to urgently purchase a microscope eyepiece adaptor to fix a Nikon >Cool Pix Digital Camera (990) on to a Lieca microscope (Leica DMR) Wetzler >Gmbh (Type 020-525-024 )in our lab. > >I would very much appreciate to know whether any one knows from where can I >get a suitable eye piece adaptor >Sincerely, > >Kushan Tennakoon >----------------------------------------------------------- > > From scoop <@t> mail.nih.gov Thu Oct 28 14:16:16 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] thanks and aquamount Message-ID: Dear Histonetters, Thanks to everyone for the great advice on bone decal and oil red o stain. I have one last question related to the oil red o stain - I just bought a bottle of aquamount to use when mounting the oil red o stained slides, but it didn't come with instructions (on the bottle it refers to instructions). Are there any special instructions for aquamount? Do I need to seal the slides with nail polish or does aquamount harden? Or, does anyone know how I can contact Lerner to get a copy of the instructions (I bought it from Fisher - Lerner doesn't answer the phone number I have for them). Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From jkiernan <@t> uwo.ca Thu Oct 28 14:21:31 2004 From: jkiernan <@t> uwo.ca (J. A. Kiernan) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] wax embedding of plant tissues with latex References: Message-ID: <418146BB.15ED941@uwo.ca> In "Plant Microtechnique and Microscopy," p.43, S.E.Ruzin says fixation in formaldehyde-acetic-alcohol is good for preserving laticifers. I suspect that this would dissolve out the latex before it can coagulate. On p.70 Ruzin recommends methacrylate embedding, and on p.143 he says laticifers can be fluorescently stained, giving a ref to Bruni & Tosi 1980. Protoplasma 102:343-347. In "Botanical Microtechnique and Cytochemistry," Berlyn & Mischke give a very detailed account of how to infiltrate and embed plant specimens but there is no mention of latex. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Kushan U Tennakoon wrote: > > Dear Histonetters, > > We have encountered problems when trying to embed Euphorbia root tissues > (that have a lot of latex) in paraffin wax. > Can you please advice me / give some fine points to ensure the optimal > penetration of paraffin wax into the plant tissues that have a lot of milky > latex. > > Sincerely, > Kushan Tennakoon > ktennako@odu.edu > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Thu Oct 28 14:40:46 2004 From: jkiernan <@t> uwo.ca (J. A. Kiernan) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Fixation of mast cells Message-ID: <41814B3E.4CF2C9E9@uwo.ca> A few days ago someone asked about fixing mast cells in rabbit tissues. (I've lost the original email.) The rabbit is one of several species with water-soluble mast cell granules. To preserve them you need to use a non-aqueous fixative such as Carnoy or an "alcoholic Bouin" mixture. When I was working on mast cells of various animals I used mainly the latter. Rats and mice have water-insoluble mast cell granules but they are also well preserved by fixation in Carnoy (which gives better micro-anatomical preservation than formaldehyde). For a short account of fixation of mast cells (with 18 references), see Hans Selye's book "The Mast Cells" (Washington: Butterworths, 1965), p.25. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ From contact <@t> excaliburpathology.com Thu Oct 28 15:16:04 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox Message-ID: <20041028201605.39960.qmail@web50310.mail.yahoo.com> Congratulations to all you Red Sox fans. My Astros were out of it awhile back. & Ian, I am going to bite my lip and not type what I am thinking about your backhanded insult to those of us in your FORMER colony. God bless Tony Blair! From neuroant <@t> hotmail.com Thu Oct 28 15:24:02 2004 From: neuroant <@t> hotmail.com (Ant S.) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Teased Nerves Message-ID: I tease nerves here, and am having a problem with the "longterm" storage. I tease them in 60% glycerin and then use that to mount them as well. That method is not a very good way to do this, but I can't add any extra media (such as aqua mount) because it makes the nerves move around on the slide, and therefore they're more difficult, or impossible, to read. I heard about possibly baking them in an oven. Does anyone out there tease? Do you have a special way of storing them? Do you bake them? And if so, for how long? Thanks in advance, Antoinette Swensson Univeristy of Washington/Harborview Medical Center Neuropathology _________________________________________________________________ [1]Rock, jazz, country, soul & more. Find the music you love on MSN Music! References 1. http://g.msn.com/8HMAENUS/2728??PS=47575 From AnthonyH <@t> chw.edu.au Thu Oct 28 18:37:57 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:24:13 2005 Subject: [Histonet] Red Sox Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E0E9@simba.kids> For what its worth, my cricket team was known as the "Red Undies". When we would "take drinks" (a term for a drink break during a game), we really knew how to party. I retired from my cricket team in order to save what was left of my liver! Congrats the Red Socks Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Angela Bitting [mailto:akbitting@geisinger.edu] Sent: Friday, 29 October 2004 3:31 AM To: amosbrooks@earthlink.net; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Red Sox Does anyone else out there think it has more to do with selling tickets, advertising time, etc. Don't you think "Baseball" knew that the Red Sox going to the W.S. would draw more of an audience than the Yankees winning again! It's all about the Benjamins! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 >>> 10/28/04 12:20PM >>> Hi, Don't say ALL of New England is happy. Here's one Connecticut Yankees fan still reeling from four consecutive losses here. I can only say the win must have had something to do with the lunar eclipse last night. Personally given the signs: war, earthquakes in Japan, an eclipse and the Red Sox winning a World Series, I'm starting work on an ark because the end must be near! Congradulations Red Sox fans your team played exceptionally and deserve the pe... penn ... pennant ** there I said it** ick I'm feeling woozy! Amos Brooks _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From am102 <@t> st-andrews.ac.uk Fri Oct 29 03:45:51 2004 From: am102 <@t> st-andrews.ac.uk (Anne-Sophie Martinez) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] (no subject) Message-ID: <200410290831.JAA27812@bute.st-andrews.ac.uk> Hello everybody!!! Thank you very much for your help. I have plenty of things to try now. AS ---------------------------------------------- Dr Anne-Sophie Martinez School of Biology, University of St. Andrews, Bute Medical Buildings, St. Andrews, KY16 9TS, UK. Tel +44 (0) 1334 463063/3365 Fax +44 (0) 1334 463600 ---------------------------------------------- From arvind <@t> nbrc.res.in Fri Oct 29 04:16:03 2004 From: arvind <@t> nbrc.res.in (Arvind Pundir) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] RE: Histonet Digest, Vol 11, Issue 37 Message-ID: <7DE0291A47C4BC4D9CDEF65D5017CEF0A6FB@mail.nbrc.res.in> " can anyone tell me the protocol for autoradiography for localising the opioid receptors in brain of avians thanks in advance Arvind NBRC, INDIA From funderwood <@t> mcohio.org Fri Oct 29 06:59:35 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:14 2005 Subject: [BULK] - RE: [Histonet] Red Sox Message-ID: Shoot Tony, don't you remember that old proverb......"The liver is evil and must be punished". >>> Tony Henwood 10/28/04 07:37PM >>> For what its worth, my cricket team was known as the "Red Undies". When we would "take drinks" (a term for a drink break during a game), we really knew how to party. I retired from my cricket team in order to save what was left of my liver! Congrats the Red Socks Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Angela Bitting [mailto:akbitting@geisinger.edu] Sent: Friday, 29 October 2004 3:31 AM To: amosbrooks@earthlink.net; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Red Sox Does anyone else out there think it has more to do with selling tickets, advertising time, etc. Don't you think "Baseball" knew that the Red Sox going to the W.S. would draw more of an audience than the Yankees winning again! It's all about the Benjamins! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 >>> 10/28/04 12:20PM >>> Hi, Don't say ALL of New England is happy. Here's one Connecticut Yankees fan still reeling from four consecutive losses here. I can only say the win must have had something to do with the lunar eclipse last night. Personally given the signs: war, earthquakes in Japan, an eclipse and the Red Sox winning a World Series, I'm starting work on an ark because the end must be near! Congradulations Red Sox fans your team played exceptionally and deserve the pe... penn ... pennant ** there I said it** ick I'm feeling woozy! Amos Brooks _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From aep10 <@t> cornell.edu Fri Oct 29 08:52:08 2004 From: aep10 <@t> cornell.edu (Anna Elisse Beaudin) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] (kinda) silly questions about IHC Message-ID: <1894.128.253.96.73.1099057928.squirrel@128.253.96.73> Hello, I have a question about IHC. I am trying to do multiple labeling on sagittal brain sections that were fixed by perfusion. I cut these sections on a cryostat, and I am having a lot of trouble deciding whether to do IHC on mounted sections vs. free-floating. When I try to mount fixed sections directly while sectioning, I get air bubbles caught between the section and the slide.. the sections just don't seem to want to lie smoothly on the slide. Alternatively, when I stain free-floating sections, these long sagittal sections curl up and as such I get extremely uneven (and ugly) staining. My actual (silly) question is whether it is possible to collect sections into solution (free-floating), mount them, let them dry overnight, and then do IHC on them. I'm torn because I need to let the section dry a little on the slide so that they stick, but at the same time I think it might be bad for them to dry out? As you can see, I am very confused, and would really appreciate anyone's advice. Thanks in advance for your help! Best, Anna Beaudin Division of Nutritional Sciences Cornell University From ftulenko06 <@t> jcu.edu Fri Oct 29 09:50:59 2004 From: ftulenko06 <@t> jcu.edu (ftulenko06@jcu.edu) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] question about air bubbles Message-ID: <2ebbea6a.96eaab1b.81e3100@mirapoint.jcu.edu> Hello everybody, I am trying to serial section museum specimens of whole turtle embryo heads. Specimens are infiltrated and embedded with paraplast X-tra. The problem I am having is that there is air trapped in the specimen. This makes the turtle head float in the molten paraplast. The areas with air are not completely infiltrated with paraplast and section poorly; the delicate tissue collapses on itself. If anybody can make any suggestions about how to remove the air and get complete infiltration, I would appreciate it. Thanks, Frank From rockbeki <@t> ufl.edu Fri Oct 29 10:15:37 2004 From: rockbeki <@t> ufl.edu (SMITH,REBEKAH FELICIA) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] (kinda) silly questions about IHC Message-ID: <1866657724.1099062937042.JavaMail.osg@osgjas01.cns.ufl.edu> I don't know if this will help, since all I know about is paraffin sections, but I mount my tissue on slides from a water bath (which will keep bubbles from popping up in between the slide and the tissue as long as the water bath isn't boiling or otherwise bubbly itself) and leave them to dry overnight on a slide warmer. It's never hurt my paraffin sections to let them dry like that, but I don't know how that compares to frozen sections. Rebekah Smith On Fri Oct 29 09:52:08 EDT 2004, Anna Elisse Beaudin wrote: > Hello, > > I have a question about IHC. I am trying to do multiple > labeling on > sagittal brain sections that were fixed by perfusion. I cut > these > sections on a cryostat, and I am having a lot of trouble deciding > whether to do IHC on mounted sections vs. free-floating. When I > try to > mount fixed sections directly while sectioning, I get air bubbles > caught between the section and the slide.. the sections just > don't seem > to want to lie smoothly on the slide. Alternatively, when I > stain > free-floating sections, these long sagittal sections curl up and > as > such I get extremely uneven (and ugly) staining. My actual > (silly) > question is whether it is possible to collect sections into > solution > (free-floating), mount them, let them dry overnight, and then do > IHC on > them. I'm torn because I need to let the section dry a little on > the > slide so that they stick, but at the same time I think it might > be bad > for them to dry out? As you can see, I am very confused, and > would > really appreciate anyone's advice. Thanks in advance for your > help! > > Best, > Anna Beaudin > Division of Nutritional Sciences > Cornell University > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" From MAUGER <@t> email.chop.edu Fri Oct 29 10:31:30 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] (kinda) silly questions about IHC Message-ID: To Anna, My experience is that it is best to dry frozen sxn slides. I used to keep them in a dessicator for weeks before staining sometimes, and then use acetone to further remove water from tissue. Jo >>> "Anna Elisse Beaudin" 10/29/04 09:52AM >>> Hello, I have a question about IHC. I am trying to do multiple labeling on sagittal brain sections that were fixed by perfusion. I cut these sections on a cryostat, and I am having a lot of trouble deciding whether to do IHC on mounted sections vs. free-floating. When I try to mount fixed sections directly while sectioning, I get air bubbles caught between the section and the slide.. the sections just don't seem to want to lie smoothly on the slide. Alternatively, when I stain free-floating sections, these long sagittal sections curl up and as such I get extremely uneven (and ugly) staining. My actual (silly) question is whether it is possible to collect sections into solution (free-floating), mount them, let them dry overnight, and then do IHC on them. I'm torn because I need to let the section dry a little on the slide so that they stick, but at the same time I think it might be bad for them to dry out? As you can see, I am very confused, and would really appreciate anyone's advice. Thanks in advance for your help! Best, Anna Beaudin Division of Nutritional Sciences Cornell University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SJones <@t> cvm.tamu.edu Fri Oct 29 10:41:26 2004 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] question about air bubbles Message-ID: Hi Frank, Is the tissue under vacuum during paraffin infiltration? Sarah Sarah Jones, HT(ASCP) Histology Lab Dept. of Veterinary Integrative Biosciences College of Veterinary Medicine Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> 10/29/2004 9:50:59 AM >>> Hello everybody, I am trying to serial section museum specimens of whole turtle embryo heads. Specimens are infiltrated and embedded with paraplast X-tra. The problem I am having is that there is air trapped in the specimen. This makes the turtle head float in the molten paraplast. The areas with air are not completely infiltrated with paraplast and section poorly; the delicate tissue collapses on itself. If anybody can make any suggestions about how to remove the air and get complete infiltration, I would appreciate it. Thanks, Frank _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hfedor <@t> jhmi.edu Fri Oct 29 10:42:17 2004 From: hfedor <@t> jhmi.edu (Helen Fedor) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] (kinda) silly questions about IHC Message-ID: If you wanted to cut free floating sections and mount them on slides and dry them over night. you can do that. I cut 30 micron sections this way and did LFB/PAS. I may have gotten down as far as 25 microns. I don't think I ever went any thinner than that on free floating sections. For Immuno the problem is you will not get the staining from both sides of the tissue. It can only penetrate through the surface and not from the side that is touching the glass. Dab is a large molecule and will not penetrate very far. That is why the Floating method is used. Double the amount of staining. Try to slow down your agitation if you can, that may help if you want to give the free floating another try. or. I have cut many frozen cryoprotected sections onto slide. start with cold slides. put the section onto the slide keeping it in the cryostat. The section is melted onto the slide very slowly using the heat from a finger on the back of the slide. Start at one end of the section. as it melts slowly slide your finger toward the frozen part(this is all on the back of the slide). While doing this use a brush on the section. and slowly brush the section down trying to keep it flat. this is all done inside the cryostat so every thing is cold. Only the heat from your finger melts the tissue It is a lot of hand and eye coordination to get this to work. But it can be done. This is done on 10 micron sections. good luck Helen >>> "SMITH,REBEKAH FELICIA" 10/29/04 11:15AM >>> I don't know if this will help, since all I know about is paraffin sections, but I mount my tissue on slides from a water bath (which will keep bubbles from popping up in between the slide and the tissue as long as the water bath isn't boiling or otherwise bubbly itself) and leave them to dry overnight on a slide warmer. It's never hurt my paraffin sections to let them dry like that, but I don't know how that compares to frozen sections. Rebekah Smith On Fri Oct 29 09:52:08 EDT 2004, Anna Elisse Beaudin wrote: > Hello, > > I have a question about IHC. I am trying to do multiple > labeling on > sagittal brain sections that were fixed by perfusion. I cut > these > sections on a cryostat, and I am having a lot of trouble deciding > whether to do IHC on mounted sections vs. free-floating. When I > try to > mount fixed sections directly while sectioning, I get air bubbles > caught between the section and the slide.. the sections just > don't seem > to want to lie smoothly on the slide. Alternatively, when I > stain > free-floating sections, these long sagittal sections curl up and > as > such I get extremely uneven (and ugly) staining. My actual > (silly) > question is whether it is possible to collect sections into > solution > (free-floating), mount them, let them dry overnight, and then do > IHC on > them. I'm torn because I need to let the section dry a little on > the > slide so that they stick, but at the same time I think it might > be bad > for them to dry out? As you can see, I am very confused, and > would > really appreciate anyone's advice. Thanks in advance for your > help! > > Best, > Anna Beaudin > Division of Nutritional Sciences > Cornell University > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Helen L. Fedor B.S. Johns Hopkins University Pathology Department 600 N Wolfe St Marburg Room 406 Baltimore MD 21287 email: hfedor@jhmi.edu Phone: 410 614-1660 Pager: 410 283-3419 WARNING: E-mail sent over the Internet is not secure. Information sent by e-mail may not remain confidential. DISCLAIMER: This e-mail is intended only for the individual to whom it is addressed. It may be used only in accordance with applicable laws. If you received this e-mail by mistake, notify the sender and destroy the e-mail. From jennifer <@t> srasf.com Fri Oct 29 11:53:13 2004 From: jennifer <@t> srasf.com (Jennifer McPhearson) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Looking for Histo. Marketing Manager Message-ID: I am currently conducting a search for a Marketing Manager, preferably with some Immunohistochemistry experience or the like. Great company, growing at 30% a year. Please email me if anyone has any suggestions, interest or referrals, jennifer@srasf.com. Thanks! Jennifer McPhearson Executive Search Consultant Sanford Rose Associates, San Francisco 1415 Oakland Blvd., Suite 215 Walnut Creek, CA 94596 Phone: (925) 974-1760 x 12 Fax: (925) 974-1763 Email: jennifer@srasf.com From ja.mitchell <@t> hosp.wisc.edu Fri Oct 29 11:19:21 2004 From: ja.mitchell <@t> hosp.wisc.edu (Mitchell Jean A.) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] (kinda) silly questions about IHC Message-ID: <583D3E9A1E843445BD54E461D1A2F6F30F83F43E@uwhis-xchng2.hosp.wisc.edu> I do IHC on 50 micron frozen sections using a free floating technique that gives great results. I use flat bottom 96 well plates for staining - it does takes a bit of practice getting the hang of transferring the sections from well to well without damaging them though. If you want more details on the staining protocol feel free to contact me! Jean Mitchell, BS, HT (ASCP) University of Wisconsin Hospital & Clinics Department of Neurology Madison, WI On Fri Oct 29 09:52:08 EDT 2004, Anna Elisse Beaudin wrote: > Hello, > > I have a question about IHC. I am trying to do multiple > labeling on > sagittal brain sections that were fixed by perfusion. I cut > these > sections on a cryostat, and I am having a lot of trouble deciding > whether to do IHC on mounted sections vs. free-floating. When I > try to > mount fixed sections directly while sectioning, I get air bubbles > caught between the section and the slide.. the sections just > don't seem > to want to lie smoothly on the slide. Alternatively, when I > stain > free-floating sections, these long sagittal sections curl up and > as > such I get extremely uneven (and ugly) staining. My actual > (silly) > question is whether it is possible to collect sections into > solution > (free-floating), mount them, let them dry overnight, and then do > IHC on > them. I'm torn because I need to let the section dry a little on > the > slide so that they stick, but at the same time I think it might > be bad > for them to dry out? As you can see, I am very confused, and > would > really appreciate anyone's advice. Thanks in advance for your > help! > > Best, > Anna Beaudin > Division of Nutritional Sciences > Cornell University > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Helen L. Fedor B.S. Johns Hopkins University Pathology Department 600 N Wolfe St Marburg Room 406 Baltimore MD 21287 email: hfedor@jhmi.edu Phone: 410 614-1660 Pager: 410 283-3419 WARNING: E-mail sent over the Internet is not secure. Information sent by e-mail may not remain confidential. DISCLAIMER: This e-mail is intended only for the individual to whom it is addressed. It may be used only in accordance with applicable laws. If you received this e-mail by mistake, notify the sender and destroy the e-mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ganglv16 <@t> hotmail.com Fri Oct 29 13:22:58 2004 From: ganglv16 <@t> hotmail.com (=?gb2312?B?wsAguNU=?=) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] IHC in mouse lung Message-ID: I have been doing IHC in mouse lung for two years. Compared with othere tissue like GI tract, it seems the backgroung staining of lung is always more stronger, no matter what Ab used. Anyone knows why and would like to give some suggestion on it? Thanks alot! Gary _________________________________________________________________ ʹÓà [1]MSN Messenger ÓëÁª»úµÄÅóÓѽøÐн»Á÷ References 1. http://g.msn.com/8HMACNCN/2731??PS=47575 From MTitford <@t> aol.com Fri Oct 29 13:46:47 2004 From: MTitford <@t> aol.com (MTitford@aol.com) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Teased Nerve time Message-ID: <127DB353.7663C978.00762DB1@aol.com> Antoinette Swensson asks about storing teased nerves that have been teased in glycerine. The newer method is to tease nerves in epoxy resin, like that used in electron microscopy. When you are finished teasing, just drop a coverslip on and you are done. The slides dry like the electron microscopy "thick sections". I don't have the reference at my fingertips but could dig it out for you if necessary. Regards Mike Titford USA Pathology Mobile AL USA From travers.3 <@t> osu.edu Fri Oct 29 16:54:09 2004 From: travers.3 <@t> osu.edu (Susan Travers) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] ImmPress Message-ID: Has anyone used the new "ImmPress" reagent from Vector? If so, do you have a feel for how it compares to ABC in terms of sensitivity? Thanks Susan Travers -- Susan Travers, Ph.D. Professor, Oral Biology The Ohio State University College of Dentistry 305 W. 12th Avenue Columbus, Ohio 43210-1267 (614)292-6366(V) (614)247-6945(F) From corporate_21 <@t> hotmail.com Fri Oct 29 14:34:21 2004 From: corporate_21 <@t> hotmail.com (corporate headquarters) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] DAKO product info Message-ID: Hi, I'm Looking for info on Dako products - no Sales reps please ! would prefer unbiased accounts of product functionality. Specifically the new Eridan system. Thanks for any help _________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar – get it now! http://toolbar.msn.click-url.com/go/onm00200415ave/direct/01/ From ajennings <@t> unmc.edu Fri Oct 29 14:56:20 2004 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] (kinda) silly questions about IHC In-Reply-To: <1894.128.253.96.73.1099057928.squirrel@128.253.96.73> Message-ID: I do float my cryo brain sections onto slides for just that reason. I have let them dry on a 50C slide warmer for two hours and had very little problem with them falling off anita "Anna Elisse Beaudin" histonet@lists.utsouthwestern.edu Sent by: cc histonet-bounces@ lists.utsouthwest Subject ern.edu [Histonet] (kinda) silly questions about IHC 10/29/2004 08:52 AM Hello, I have a question about IHC. I am trying to do multiple labeling on sagittal brain sections that were fixed by perfusion. I cut these sections on a cryostat, and I am having a lot of trouble deciding whether to do IHC on mounted sections vs. free-floating. When I try to mount fixed sections directly while sectioning, I get air bubbles caught between the section and the slide.. the sections just don't seem to want to lie smoothly on the slide. Alternatively, when I stain free-floating sections, these long sagittal sections curl up and as such I get extremely uneven (and ugly) staining. My actual (silly) question is whether it is possible to collect sections into solution (free-floating), mount them, let them dry overnight, and then do IHC on them. I'm torn because I need to let the section dry a little on the slide so that they stick, but at the same time I think it might be bad for them to dry out? As you can see, I am very confused, and would really appreciate anyone's advice. Thanks in advance for your help! Best, Anna Beaudin Division of Nutritional Sciences Cornell University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cuttingedgehistology <@t> yahoo.com Fri Oct 29 18:32:29 2004 From: cuttingedgehistology <@t> yahoo.com (Brandon Stokes) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Antigen Retrieval w/ B&D Steamer..... Message-ID: <20041029233229.70857.qmail@web41407.mail.yahoo.com> Hi fellow Histonetters........ I have a quick question for anyone out there that knows anything about using a Black&Decker Steamer for Antigen Retrieval. How long do you pre-heat the solutions? How long do you leave in the steamer? How long do you cool down? Those are a few of the questions that I have so far......If there is any other helpful advice using this method it would be greatly appreciated. Thanks, Brandon Stokes, HT(ASCP) Lab Manager Cutting Edge Histology Services, LLC Tigard, OR 503-443-2157 --------------------------------- Do you Yahoo!? Take Yahoo! Mail with you! Get it on your mobile phone. From kmerriam2003 <@t> yahoo.com Fri Oct 29 19:13:59 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Antigen Retrieval w/ B&D Steamer..... In-Reply-To: <20041029233229.70857.qmail@web41407.mail.yahoo.com> Message-ID: <20041030001359.309.qmail@web52507.mail.yahoo.com> Hello, This is what I do: 1. Preheat retrieval solution in microwave for 6 minutes (3 tissue tek containers will fit into 1 steamer) 2. Preheat steamer for 10-15 minutes (you can do this while bringing slides down to water). 3. Heat slides for 45-60 minutes 4. Cool down for at least 10 minutes Kim Merriam Novartis Cambridge, MA Brandon Stokes wrote: Hi fellow Histonetters........ I have a quick question for anyone out there that knows anything about using a Black&Decker Steamer for Antigen Retrieval. How long do you pre-heat the solutions? How long do you leave in the steamer? How long do you cool down? Those are a few of the questions that I have so far......If there is any other helpful advice using this method it would be greatly appreciated. Thanks, Brandon Stokes, HT(ASCP) Lab Manager Cutting Edge Histology Services, LLC Tigard, OR 503-443-2157 --------------------------------- Do you Yahoo!? Take Yahoo! Mail with you! Get it on your mobile phone. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Yahoo! Mail – CNET Editors' Choice 2004. Tell them what you think. From kmerriam2003 <@t> yahoo.com Fri Oct 29 19:18:29 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Offtopic - Microscope for kids Message-ID: <20041030001829.17947.qmail@web52510.mail.yahoo.com> Hello all, My 8 year old son wants a microscope for Christmas. I have been searching on the internet for a decent one that does not cost an arm and a leg. It seems like there are lots of cheap ones in the $12-30 range and then some nice ones that are in the $100-200 range. Does anyone know where I could get one that costs around $50-70 that is not a piece of junk? I would like 2 or 3 objectives. Of course, the slides need not be included! Thanks in advance, Kim Merriam Novartis Cambridge, MA Kim Merriam Novartis Cambridge, MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From jkiernan <@t> uwo.ca Sat Oct 30 00:48:17 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Offtopic - Microscope for kids References: <20041030001829.17947.qmail@web52510.mail.yahoo.com> Message-ID: <41832B21.F17900CD@uwo.ca> WHATEVER YOU DO: Don't buy one without first having a look at something with it yourself. Even in the highest of the price ranges you mention the optics cannot be of high quality, so there's no point getting anything with an objective that's more than X10. The mechanical parts are also likely to be stiff, loose, or otherwise not easy to control. The best value for money is a 2nd-hand student microscope. University departments often sell these off for $100 or less, and you'd be lucky to get an equivalent new instrument for $1000. An old student microscope would not impress your 8 year-old son, because it would need too many adjustments (condenser, brightness, centering, positioning the slide etc). My experience with children and microscopes is that those younger than about 13 find it difficult to keep an eye steadily in place over an eyepiece - not too far away (nothing visible), not too close (disturbing images of reflected bent eyelashes), and not wobbling either. Children need help finding something to look at because moving the slide is another difficulty (especially if you don't have a mechanical stage, which is 2 more knobs). Focusing is a must, of course, and using coarse and fine knobs can easily be taught. >From my experiences with children (including me, my friends long ago, my 5 grown-up kids, and some of our grandchildren), you need to be 16 to handle a real microscope. Not everyone would agree. The Royal Microscopical Society (in Britain) has been campaigning for "a microscope in every school" for several years. "Every school" means elementary schools, which are for under-11s in the UK. The RMS has identified various kinds of microscope appropriate for children. They have a major web site, http://www.rms.org.uk Click on the Education tab, then on AMFES to get their advice on paedomicrophilia. Consider buying your son a X10 lens and showing him how to see a spider's fangs or the stinging hairs of a nettle. Hope this helps! John Kiernan London, Canada. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Kim Merriam wrote: > > Hello all, > > My 8 year old son wants a microscope for Christmas. I have been searching on the internet for a decent one that does not cost an arm and a leg. It seems like there are lots of cheap ones in the $12-30 range and then some nice ones that are in the $100-200 range. Does anyone know where I could get one that costs around $50-70 that is not a piece of junk? I would like 2 or 3 objectives. Of course, the slides need not be included! > > Thanks in advance, > Kim Merriam > Novartis > Cambridge, MA > > Kim Merriam > Novartis > Cambridge, MA > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Ianbernard <@t> netzero.com Sat Oct 30 06:04:02 2004 From: Ianbernard <@t> netzero.com (Ian) Date: Fri Sep 16 15:24:14 2005 Subject: [BULK] - [Histonet] Red Sox In-Reply-To: Message-ID: Go Astros!!!!!!!!!! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Thursday, October 28, 2004 6:27 AM To: Jackie M O'Connor; Fred Underwood Cc: histonet@lists.utsouthwestern.edu; BMolinari@heart.thi.tmc.edu; histonet-bounces@lists.utsouthwestern.edu Subject: RE: [BULK] - [Histonet] Red Sox Jackie, I'm sorry, Fred will be tied up next year (even if I have to do it myself) Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jackie M O'Connor Sent: Thursday, October 28, 2004 7:25 AM To: Fred Underwood Cc: histonet@lists.utsouthwestern.edu; BMolinari@heart.thi.tmc.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [BULK] - [Histonet] Red Sox Fred, have you ever been to Wrigley? My daughter works there - she'll be glad to give you a tour if you can guarantee the same results. "Fred Underwood" Sent by: histonet-bounces@lists.utsouthwestern.edu 10/28/2004 07:12 AM To: , cc: Subject: Re: [BULK] - [Histonet] Red Sox I'm not one to toot my own horn, but. I made my first trip to Boston 2 weekends ago, which included a tour of Fenway. The next thing you know the Sox win it all. Do you really think it's a coincidence? No thanks are necessary. Just remember the good karma of the Frederoo will trump the curse of the Bambino everytime. >>> "Molinari, Betsy" 10/28/04 07:23AM >>> Good morning, yes it is! Thank you Red Sox! I hope no one minds my posting but I came in to the lab this morning on cloud 9 and had to share ! Betsy Molinari HT (ASCP) Texas Heart Institute Houston (but born and raised in Boston :-)) TX 832-355-6524 _______________________________________________ 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From msdfloyd <@t> worldnet.att.net Sat Oct 30 08:28:35 2004 From: msdfloyd <@t> worldnet.att.net (Dina Floyd) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Antigen Retrieval w/ B&D Steamer..... References: <20041029233229.70857.qmail@web41407.mail.yahoo.com> Message-ID: <001201c4be84$5c2ed4b0$a68c6520@yourxhtr8hvc4p> This is how we do it... If you are using Citra, preheat Citra for 30 minutes, you can do this while your slides are running down to water. Put slides in for 20-30 minutes, Cool down to 40 degrees before rinsing in APK. If you are using Trilogy, we start with two coplin jars of trilogy. Place your slides in the first jar as soon as they come out of the oven (DO NOT RUN DOWN TO WATER) Place coplin jars side by side in steamer and heat for 30 minutes. Remove slides from first trilogy and transfer to second trilogy that has been preheating beside first trilogy. Steam another 30 minutes. Remove slides and immediately wash in several changes of apk wash. -- Original Message ----- From: "Brandon Stokes" To: Sent: Friday, October 29, 2004 6:32 PM Subject: [Histonet] Antigen Retrieval w/ B&D Steamer..... > Hi fellow Histonetters........ > > I have a quick question for anyone out there that knows anything about > using a Black&Decker Steamer for Antigen Retrieval. > > How long do you pre-heat the solutions? > How long do you leave in the steamer? > How long do you cool down? > > Those are a few of the questions that I have so far......If there is any > other helpful advice using this method it would be greatly appreciated. > > Thanks, > > Brandon Stokes, HT(ASCP) > Lab Manager > Cutting Edge Histology Services, LLC > Tigard, OR > 503-443-2157 > > > --------------------------------- > Do you Yahoo!? > Take Yahoo! Mail with you! Get it on your mobile phone. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From JMcCormi <@t> schosp.org Sat Oct 30 08:33:45 2004 From: JMcCormi <@t> schosp.org (McCormick, James) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Offtopic - Microscope for kids Message-ID: <229A3566B9F0D311826E00D0B7441D7905E3012B@swedish_nt1.schosp.org> Kim, You have received some good advice from John Kiernan. Learning to use a hand lens to bring "little things" closer to view is the best building block for "experimenting" with the magnification of interesting objects. Someone has said "we need a low power lens and a high power mind!" There are some interesting books and web based assets to build with. From my experience it is important that the beginner will benefit greatly from "examining" little things with a hand lens and making drawings of what they see. If you look up "bugs" on Google it will lead you to www.BillyBear4Kids.com I think you will find some things of great interest to an 8 year old. Wait until he is 12 to spend $100.00 for a used student microscope. Good Luck, I am a pathologist and have used a microscope since age 12. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Sent: Saturday, October 30, 2004 12:48 AM To: Kim Merriam; Histonet Subject: Re: [Histonet] Offtopic - Microscope for kids WHATEVER YOU DO: Don't buy one without first having a look at something with it yourself. Even in the highest of the price ranges you mention the optics cannot be of high quality, so there's no point getting anything with an objective that's more than X10. The mechanical parts are also likely to be stiff, loose, or otherwise not easy to control. The best value for money is a 2nd-hand student microscope. University departments often sell these off for $100 or less, and you'd be lucky to get an equivalent new instrument for $1000. An old student microscope would not impress your 8 year-old son, because it would need too many adjustments (condenser, brightness, centering, positioning the slide etc). My experience with children and microscopes is that those younger than about 13 find it difficult to keep an eye steadily in place over an eyepiece - not too far away (nothing visible), not too close (disturbing images of reflected bent eyelashes), and not wobbling either. Children need help finding something to look at because moving the slide is another difficulty (especially if you don't have a mechanical stage, which is 2 more knobs). Focusing is a must, of course, and using coarse and fine knobs can easily be taught. >From my experiences with children (including me, my friends long ago, my 5 grown-up kids, and some of our grandchildren), you need to be 16 to handle a real microscope. Not everyone would agree. The Royal Microscopical Society (in Britain) has been campaigning for "a microscope in every school" for several years. "Every school" means elementary schools, which are for under-11s in the UK. The RMS has identified various kinds of microscope appropriate for children. They have a major web site, http://www.rms.org.uk Click on the Education tab, then on AMFES to get their advice on paedomicrophilia. Consider buying your son a X10 lens and showing him how to see a spider's fangs or the stinging hairs of a nettle. Hope this helps! John Kiernan London, Canada. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Kim Merriam wrote: > > Hello all, > > My 8 year old son wants a microscope for Christmas. I have been searching on the internet for a decent one that does not cost an arm and a leg. It seems like there are lots of cheap ones in the $12-30 range and then some nice ones that are in the $100-200 range. Does anyone know where I could get one that costs around $50-70 that is not a piece of junk? I would like 2 or 3 objectives. Of course, the slides need not be included! > > Thanks in advance, > Kim Merriam > Novartis > Cambridge, MA > > Kim Merriam > Novartis > Cambridge, MA > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > 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 *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. From asmith <@t> mail.barry.edu Sat Oct 30 11:24:14 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Offtopic - Microscope for kids Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3C21@exchsrv01.barrynet.barry.edu> I see no problem in getting a microscope for an 8 year old. Some kids are precocious. A good $30 microscope will have only an 8X or 10X lens (in other words, it will be a souped-up magnifying glass). My daughter liked to use one for observing freshwater invertebrates when she was 9. A $30 microscope with anything more will be junk. Carolina Biological supply has a decent microscope (RG-59-1205) for $125. It is the best I've seen for under $200. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Merriam Sent: Friday, October 29, 2004 8:18 PM To: Histonet Subject: [Histonet] Offtopic - Microscope for kids Hello all, My 8 year old son wants a microscope for Christmas. I have been searching on the internet for a decent one that does not cost an arm and a leg. It seems like there are lots of cheap ones in the $12-30 range and then some nice ones that are in the $100-200 range. Does anyone know where I could get one that costs around $50-70 that is not a piece of junk? I would like 2 or 3 objectives. Of course, the slides need not be included! Thanks in advance, Kim Merriam Novartis Cambridge, MA Kim Merriam Novartis Cambridge, MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From ftulenko06 <@t> jcu.edu Sat Oct 30 16:54:54 2004 From: ftulenko06 <@t> jcu.edu (ftulenko06@jcu.edu) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Thanks for the suggestions about air buubles Message-ID: <38740e96.97954f90.81a1500@mirapoint.jcu.edu> I have not been using a vacuum during processing, but will try it. I will also extend the clearing times. Thanks for help. Frank From peoshel <@t> wisc.edu Sun Oct 31 10:46:36 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Offtopic - Microscope for kids In-Reply-To: <20041030001829.17947.qmail@web52510.mail.yahoo.com> References: <20041030001829.17947.qmail@web52510.mail.yahoo.com> Message-ID: Kim, Check out the Project Micro information on the Microscopy Society of America website, http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html They (well, Caroline Schooley, really) go into just this issue -- which are good microscopes for kids and students. You can also get lots of information about activities, books, and so forth at this site. Phil >Hello all, > >My 8 year old son wants a microscope for Christmas. I have been >searching on the internet for a decent one that does not cost an arm >and a leg. It seems like there are lots of cheap ones in the $12-30 >range and then some nice ones that are in the $100-200 range. Does >anyone know where I could get one that costs around $50-70 that is >not a piece of junk? I would like 2 or 3 objectives. Of course, >the slides need not be included! > >Thanks in advance, >Kim Merriam >Novartis >Cambridge, MA > > >Kim Merriam >Novartis >Cambridge, MA -- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From haldana <@t> unimoron.edu.ar Sun Oct 31 19:13:36 2004 From: haldana <@t> unimoron.edu.ar (Hernan Aldana) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] HYALURONIC ACID detection In-Reply-To: <20041004140231.SM01372@swlx162.swmed.edu> Message-ID: <000e01c4bfb0$31ec3710$389559c8@b3w6zzmtb6juvbs> Dears histonetters I like to demonstrate the presence of hyaluronic acid in fixed eyes. Can anybody help me with some advices? Histochemical or immunohistochemical. Please help me. Thanks in advance to all Dr. Hern?n J. Aldana Marcos Facultad de Medicina. Universidad de Mor?n Machado 914. B1708JPD. Buenos Aires. Argentina e-mail alternativo hernanjavier@yahoo.com web: http://hjaldanamarcos.bravepages.com http://www.ht.org.ar From jkiernan <@t> uwo.ca Sun Oct 31 21:26:38 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] HYALURONIC ACID detection References: <000e01c4bfb0$31ec3710$389559c8@b3w6zzmtb6juvbs> Message-ID: <4185ACEE.CF1AFC0B@uwo.ca> Some properties of hyaluronic acid. (1) Stains with alcian blue 8G at pH 2.5 but not at pH 1.0. (2) Staining prevented by prior treatment with streptococcal hyaluronidase. (Testicular hyaluronidase is cheaper & more easily available but can also remove chondroitin sulphates. Chondroitin sulphates are stained by alcian blue at both pH 1 and 2.5. (3) Staining by alcian blue is reversibly blocked by prior methylation of the section. Stainability is restored if methylation is followed by saponification. The staining of chondroitin sulphates with alcian blue is irreversibly blocked by methylation. (4) Negative with classical PAS method but becomes PAS-positive if a special oxidation sequence is used (Scott & Harbinson 1969 Histochemie 19:155-161). If you have pure hyaluronic acid there will be no difficulty staining it. If, as is usually the case, other macromolecular carbohydrates or glycoproteins (collagen, for example) are present in exactly the same place, you'll need to do all the above methods and more. The best account of carbohydrate histochemistry that I know is Chapter 15 in Vol 2 of Pearse's "Histochemistry". It's quite heavy going, but this is a field where you really do need to be in command of every step of every method. Hope this helps. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Hernan Aldana wrote: > > Dears histonetters > I like to demonstrate the presence of hyaluronic acid in fixed eyes. Can > anybody help me with some advices? > Histochemical or immunohistochemical. Please help me. > Thanks in advance to all > > Dr. Hern?n J. Aldana Marcos > Facultad de Medicina. Universidad de Mor?n > Machado 914. B1708JPD. Buenos Aires. Argentina > e-mail alternativo hernanjavier@yahoo.com > web: http://hjaldanamarcos.bravepages.com > http://www.ht.org.ar > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet