From jhnspam <@t> aol.com Sat May 1 22:23:25 2004 From: jhnspam <@t> aol.com (jhnspam@aol.com) Date: Fri Sep 16 15:22:53 2005 Subject: [Histonet] nail sectioning Message-ID: <85.acd226e.2dc5c3ad@aol.com> We face in to the block and then soak it in Downy Fabric Softener for about 5 minutes. Put the section on a plus slide and it comes out beautiful. We do PAS/LG on all nails and they look great as well. Pam Johnson From Stanley.Stylli <@t> mh.org.au Sun May 2 21:53:02 2004 From: Stanley.Stylli <@t> mh.org.au (Stylli, Stanley) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] ssDNA Message-ID: <00519F9F41D2844D9C75BEE1F6AC09AF01992994@rmhmail1.ssg.org.au> Dear Histonetters, Has anyone come across and antibody to ssDNA other than the one from Alexis Biochemicals (F7-26) Thanks Stan From Megan.Kear <@t> hunter.health.nsw.gov.au Mon May 3 01:05:08 2004 From: Megan.Kear <@t> hunter.health.nsw.gov.au (Megan Kear) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] laminin response Message-ID: thank you to all for the response to my request for a polyclonal laminin antibody. What service! Thank you Megan Clarke ( Nee Kear) HAPS Newcastle Australia From David.Edmondson <@t> christie-tr.nwest.nhs.uk Mon May 3 04:43:35 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Mounting media Message-ID: Hi DAKOs Faramount is fine with their BCIP/NBT/INT ( S3025 ) Dave Christie H Manchester -----Original Message----- From: Patti Loykasek [mailto:ploykasek@phenopath.com] Sent: 29 April 2004 17:45 To: histonet Subject: [Histonet] Mounting media Thanks for all the help with the BCIP/NBT question. Now I have another question from the same research coworker. She would like to use an aqueous based mounting media. The one we used to use is no longer available. What is a good one to purchase for this purpose? Thanks again. Patti Loykasek PhenoPath Laboratories Seattle, WA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From srishan <@t> mail.holyname.org Mon May 3 05:24:58 2004 From: srishan <@t> mail.holyname.org (srishan@mail.holyname.org) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Disposal of alcohols Message-ID: Hello everyone, Can anyone help me out, with the procedure for disposing alcohols from the tissue processors and stainers? Apparently there is a regulation by EPA that alcohols cannot be poured down the sink. Thank you in advance Mala Srishan From balajimr <@t> drreddys.com Mon May 3 06:14:05 2004 From: balajimr <@t> drreddys.com (balajimr@drreddys.com) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Website for histopathology of rat and mouse Message-ID: Could anyone please let me know a website (like Histonet) for exchange of information pertaining to (toxicologic ) histopathology of Rat and mouse? Thanks in advance Dr. Balaji From balajimr <@t> drreddys.com Mon May 3 06:20:02 2004 From: balajimr <@t> drreddys.com (balajimr@drreddys.com) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Testes Bouin's fluid Message-ID: Dear Histonetter, We have been preserving testes (of rats and mouse) in bouin's solution and after 24 hrs washing them with 70% alcohol and fixing it back in 10%NBF. But staining later with H & E is not that great. Can anybody suggest me the best protocol for fixing,processing and staining of testes. Dr. Balaji Dept. Pre clinical safety evaluation, Discovery research, Dr. Reddys Laboratories ltd. Bollaram Road, Miyapur, Hyderabad, 500 049 Andhra Pradseh, INDIA From StarkusL <@t> ummhc.org Mon May 3 06:49:00 2004 From: StarkusL <@t> ummhc.org (Starkus, Laurie) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Embedding in OCT and subsequent steps Message-ID: Actually, that's not completely true. Surgipath makes a mountant/clearing agent in one called Clearium. I use it instead of xylene or xylene substitute. Although, I guess in the strictest sense of the word, it is a "substitute". -----Original Message----- From: Charles Scouten [mailto:cwscouten@myneurolab.com] Sent: Friday, April 30, 2004 2:52 PM To: degaboh@rice.edu; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Embedding in OCT and subsequent steps Xylene infiltrates tissue, and prepares it for coverslipping (ie, with Permount). You still need xylene or a substitute if you plan to coverslip 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 ZP Sent: Friday, April 30, 2004 12:08 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Embedding in OCT and subsequent steps If I embed my sample in OCT, do I need to use xylene in later steps (i.e. when staining or before mounting)? I have a protocol that does that but I thought xylene was really more for paraffin embedding. Thanks!! Zarana Patel degaboh@rice.edu _______________________________________________ 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 RRushing <@t> pathgroupla.com Mon May 3 07:29:30 2004 From: RRushing <@t> pathgroupla.com (Rushing, Roxane) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] RE: Histonet Digest, Vol 5, Issue 47 Message-ID: Please send -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Friday, April 30, 2004 11:47 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 5, Issue 47 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. Re: IHC camera and scope set up? (Robert Schoonhoven) 2. Re: anti-CD45 antibody for mouse parrafin sections (Gayle Callis) 3. PA Job Descriptions (Amy Self) 4. RE: PA Job Descriptions (Gary Gill) 5. re anti laminin Ab reagent (Carl) 6. Re: Finding Plain Slides (yangpw@umich.edu) 7. grinding bone (Histology) 8. Microscope parts (Bill Blank) 9. NSH Veterinary, Industry and Research Committee webpage update (Gayle Callis) 10. Re: Microscope parts (mari.ann.mailhiot@leica-microsystems.com) 11. nail sectioning (Bliss, Mary E.) 12. grinding bone (Histology) 13. Arizona Society conference (Karen Lahti) 14. Re: water bath policy (KAELPERS@aol.com) 15. Re: slide stainers - vendors (KAELPERS@aol.com) 16. Tissue array (yan gao) 17. RE: quickest freezing method AND maintain morphology of mouse brains for LCM then microarray??? (Kemlo) 18. RE: nail sectioning (Thom Jensen) 19. Re: nail sectioning (mark.lewis@thermo.com) 20. RE: laminin (GUTIERREZ, JUAN) 21. Looking for ZAP-70and CD38 antibodies (Nava, Josefa) 22. Container (Rebecca Barnhart) 23. RE: Container (Gladney, Diane C Ms MACH) 24. RE: Job interview questions (Kari Bradshaw) 25. Contamination of processor alcohols (White, Lori) 26. RE: Container (Greer, Patricia) 27. RE: Zinc formalin and 10% NBF (Smith, Allen) 28. Re: thick sections in zebrafish (Don.Birgerson@leica-microsystems.com) 29. Re: Tissue array (Scott Mordue) ---------------------------------------------------------------------- Message: 1 Date: Thu, 29 Apr 2004 13:08:13 -0400 From: Robert Schoonhoven Subject: Re: [Histonet] IHC camera and scope set up? To: jason madore Cc: histonet@pathology.swmed.edu Message-ID: <4091367D.1040804@email.unc.edu> Content-Type: text/plain; charset=us-ascii; format=flowed This is a question with MANY answers and all of them have merit. For my light microscope I have an Olympus BH-2 with a Qimaging Micropulisher 5.0camera. My inverted Flou. microscope is an Olympus IX 71 with the Olympus MicroFire 5 MP camera. I also have it setup so that I can move the MicroFire to a second (light) microscope. Both of these cameras are 5 megapixil cameras the MicroFire will take 16 bit images and the Qimaging takes 12 bit images. In addition to the digital cameras ( well actually I had it before the digitals') we have a dedicated Olympus setup for traditional film. Ok, that tells you what I have but not the really important stuff - like why??? I'll try to make this short but there is a lot of information to take into consideration (which means that I will skip over a few things). First one needs to take into account the available funds and where to allocate them within your proposed system, as well as whether to go digital or stay with the traditional film type of system. Funds - spend as much as possible on the optics of the microscope, the best camera in the world will not correct for bad glass (optics). It really is a matter of user prefferance as to what the make of the scope is but do get the best objectives that your budget will allow for. Film vs Digital - As good as digital is with today's technology (16 MP + cameras are available), film still gives the best resolution. That said.... it's pretty darn hard to tell the difference between the two when looking at a printed micrograph in a publication. We've been submitting digital micrographs for several years with no complaints. Archived film will last for centuries , digital storage media does not. Photomicroscopy using a film camera actually takes a fair amount of skill and knowledge without even going into the darkroom part of it, where as I can show a student how to take a digital image in less then an hour and have them get good micrographs each time (I said good, not great). It should also be noted that while the old saying is "that film is cheap take lots of pictures", electrons are cheaper take more". The preceding is true but................ the initial cost of a dedicated digital camera, software and computer is MUCH higher than that of a dedicated film camera. Another good point is that the computer is your darkroom for digital images and there are incredible things that you can do with programs like Adobe Photoshop. I could go on for many pages regarding the other important factors like camera bit depth (8, 12, & 16), image analysis printing, software, ad nausium but I'm running out of time. Bob, Robert Schoonhoven, UNC-CH jason madore wrote: > Can anyone recommend a reasonable camera/scope/computer setup for > taking microphotographs of standard IHC slides and TMA slides? > > _________________________________________________________________ > Tired of spam? Get advanced junk mail protection with MSN Premium > http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=htt p://hotmail.com/enca&HL=Market_MSNIS_Taglines > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 2 Date: Thu, 29 Apr 2004 11:44:33 -0600 From: Gayle Callis Subject: Re: [Histonet] anti-CD45 antibody for mouse parrafin sections To: "m. van mkempen" , Histonet@lists.utsouthwestern.edu Message-ID: <3.0.6.32.20040429114433.00bc8cf8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii" I suggest you buy the rat antiMouse anti CD45 monoclonal, the one you are attempting to work with detects CD45 in rat tissues. BD Pharmingen carries the rat antiMouse. I am not sure if the OX-1 clone even cross reacts to mouse antigens, plus you will have mouse to mouse problems. Any clone with OX-1 is usually a rat clone. At 04:14 PM 4/29/2004 +0200, you wrote: >Hello, > >Are there any people who have experience with the following anti-CD45 >antibody: mouse monoclonal, clone OX-1 from BD #550566. > >I am trying to get this antibody to work on MOUSE spleen parrafin >sections (It works in RAT cryosections). The tissue has been fixed in 4% >paraformaldehyde during 24h at 4 C. After making the sections (5-6 >microns) and deparrafinizing I did the following steps: > >Endogenous peroxidase blocking with 3% H2O2 in methanol for 20 min >Rinse with PBS >HIER with a citrate buffer (pH 6,0 and microwave 450W, 9 min) >Rinse with PBS >Block with PBS 5%BSA (30 min, RT) >Incubate with primairy antibody dilluted 1:50 in PBS 5%BSA at (O/N, 4 C) >Rinse with PBS >Incubate with rabbit anti-mouse immunoglbulins (DAKO #Z 0259) dilluted >1:50 in PBS 5%BSA (30min, RT) >Rinse with PBS >Incubate with monoclonal mouse PAP (DAKO #P 0850) dilluted 1:50 in PBS >5%BSA (30min, RT) >Rinse with PBS > >Enzyme detection with DAB substrate (5-7 min) > >Afterwards I can not detect anything besides some background from the >endogenous peroxidase. Should I try another antibody or can I improve my >protocol? > >Best regards, Marion van Kempen (Erasmus MC, Rotterdam The Netherlands) > > > > > > > >-- >---------------------------- >*- mkempen -* >MAILTO:m.vankempen@erasmusmc.nl >----------------------------------------- > >_______________________________________________ >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: 3 Date: Thu, 29 Apr 2004 13:51:35 -0400 From: "Amy Self" Subject: [Histonet] PA Job Descriptions To: Message-ID: <39836CD6DB61654E8F95A35898C921860A86CF@exchange.gmhpost.com> Content-Type: text/plain; charset="iso-8859-1" Dear Histonetters, I need your help again......... Does anyone have a "Pathologist Assistant Job Description" that they would be willing to share with me..... Thanks in advance....amy Amy Self Georgetown Hospital Systems 843-527-7179 (phone) 843-520-7882 (fax) NOTE: The information contained in this message may be privileged, 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 this message and deleting it from your computer. Thank you. ------------------------------ Message: 4 Date: Thu, 29 Apr 2004 13:05:18 -0500 From: Gary Gill Subject: RE: [Histonet] PA Job Descriptions To: 'Amy Self' , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain Go to: http://www.pathologistsassistants.org/Public_content/General_info/what%20is. htm -----Original Message----- From: Amy Self [mailto:ASelf@gmhsc.com] Sent: Thursday, April 29, 2004 12:52 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PA Job Descriptions Dear Histonetters, I need your help again......... Does anyone have a "Pathologist Assistant Job Description" that they would be willing to share with me..... Thanks in advance....amy Amy Self Georgetown Hospital Systems 843-527-7179 (phone) 843-520-7882 (fax) NOTE: The information contained in this message may be privileged, 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 this message and deleting it from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Thu, 29 Apr 2004 20:08:35 +0100 From: "Carl" Subject: [Histonet] re anti laminin Ab reagent To: Message-ID: <002301c42e1d$5f66abf0$82412cd9@home> Content-Type: text/plain; charset="iso-8859-1" Sigma L9393 polyclonal. For me, works on human, mouse, chick, with proteolytic or M/W pretreatment ( I prefer M/W; less variable). --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.668 / Virus Database: 430 - Release Date: 24/04/2004 ------------------------------ Message: 6 Date: Thu, 29 Apr 2004 15:12:30 -0400 From: yangpw@umich.edu Subject: Re: [Histonet] Finding Plain Slides To: histonet@pathology.swmed.edu Message-ID: <1083265950.4091539e2fb50@mail.umich.edu> Content-Type: text/plain; charset=ISO-8859-1 Hi, Usually I use superfrost for slide-mounted sections. And for free floating IHC sections, I buy Gold Seal precleaned slide from Fishersci (cat # 3064) and gel coat them. Pengwei Yang University of Michigan ------------------------------ Message: 7 Date: Thu, 29 Apr 2004 13:09:34 -0700 From: "Histology" Subject: [Histonet] grinding bone To: Message-ID: <000d01c42e25$e53f1200$fda55742@ca.sprintbbd.net> Content-Type: text/plain; charset="iso-8859-1" Does anybody have a procedure for grinding bone on a buehler ecomet-3. Thank you connie ------------------------------ Message: 8 Date: Thu, 29 Apr 2004 15:32:15 -0500 From: Bill Blank Subject: [Histonet] Microscope parts To: histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" Does anyone know where I can get a halogen lamp socket for A Reichert Microstar IV scope? TIA, Bill -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board ------------------------------ Message: 9 Date: Thu, 29 Apr 2004 15:41:44 -0600 From: Gayle Callis Subject: [Histonet] NSH Veterinary, Industry and Research Committee webpage update To: Histonet@lists.utsouthwestern.edu Message-ID: <3.0.6.32.20040429154144.00bc6a30@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii" Dear All, The National Society for Histotechnology, VIR committee has an updated their webpage. Go to www.NSH.org and click on VIR. Updates are: Corrections to the Animal Processing Manual's first edition are posted for those of you who purchased the manual from NSH. Message from the new chairman of VIR Committee. Contact information Watch for VIR related Literature and Links and comments from chairman of Hard Tissue Committee. A call to all VIR histotechnologists to join the committee (membership form available on the VIR page) and update member personal information. Gayle Callis 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) 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: 10 Date: Thu, 29 Apr 2004 16:43:30 -0500 From: mari.ann.mailhiot@leica-microsystems.com Subject: Re: [Histonet] Microscope parts To: Bill Blank Cc: histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; charset=us-ascii Bill Please contact Mark Streebel at 716 686 3166. He should be able to help you. Regards Mari Ann Mailhiot BA HT ASCP Application Specialist Leica Technical Assistance Center 800 248 0123 x7267 847 236 3063 fax mari.ann.mailhiot@leica-microsystems.com www.leica-microsystems.com Bill Blank To: histonet@pathology.swmed.edu Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] Microscope parts western.edu 04/29/2004 03:32 PM Does anyone know where I can get a halogen lamp socket for A Reichert Microstar IV scope? TIA, Bill -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Thu, 29 Apr 2004 15:07:08 -0700 From: "Bliss, Mary E." Subject: [Histonet] nail sectioning To: Message-ID: <27066863AD02214B81FE49477F3E71AD0D6F2C3C@hinet1.hinet.org> Content-Type: text/plain; charset="us-ascii" Hi to All, Does anyone have a good procedure for fixing and cutting in good fingernail and toenail sections? We are trying to come up with something which will allow us to produce a good stainable slide with ALL of the tissue still intact on it! I have heard about using trichloroacetic acid but can't seem to find a procedure anywhere. Have also heard about /tried Nair and soapy water. Does anyone have any information to share? Humbly yours, Mary E. Bliss Lead Histologist Northwest Pathology, P.S. 3614 Meridian St. Suite 100 Bellingham, WA 98225 (360)734-2800 x601 (360)734-3818 FAX ------------------------------ Message: 12 Date: Thu, 29 Apr 2004 16:04:40 -0700 From: "Histology" Subject: [Histonet] grinding bone To: Message-ID: <000d01c42e3e$5b558d80$fda55742@ca.sprintbbd.net> Content-Type: text/plain; charset="iso-8859-1" Sorry, I'am processing the bones and embedding in (MMA) Methy methacrylate. thanks connie ------------------------------ Message: 13 Date: Thu, 29 Apr 2004 17:19:31 -0700 From: "Karen Lahti" Subject: [Histonet] Arizona Society conference To: "histonet" Message-ID: Content-Type: text/plain; charset="iso-8859-1" The Arizona Society for Histotechnology and Region VII conference is planned for June 3-5, 2004 in Phoenix, Arizona. The complete booklet along with registration and membership forms can be obtained from our website - www.arizonahisto.org. We have many workshop planned to include PCR, Forensic Anthropology, HT Readiness, QIHC, H&E's from A to Z, IHC, Chemical Hazards, ISH, Yoga, Infectious Diseases and TEM/SEM. Please Do Not submit funds online. The forms are available to be downloaded then printed off. Please contact me if you have any problems or questions. We are having a drawing for a free room for three nights for paid registrants by May 10, 2004. So far, we have 22 vendors coming to support our society. The Best Western Grace Inn in Ahwatukee has free airport shuttle and is providing great room rates - $59/night for a single up to $79/night for a quad. Hospitality suites and one-bedroom suites are available. www.graceinn.com for more information. Thank You, Karen Lahti ASH President ------------------------------ Message: 14 Date: Thu, 29 Apr 2004 21:04:12 EDT From: KAELPERS@aol.com Subject: Re: [Histonet] water bath policy To: nlouisea@gis.net, Histonet@lists.utsouthwestern.edu Message-ID: <78.55a0f4c3.2dc3000c@aol.com> Content-Type: text/plain; charset="US-ASCII" I have 2 references that mention cleaning the water bath, however the word cross contamination is not mentioned. If I find something I will send it to you - Good Luck with your CAP inspection! Theory and Practice of Histotechnology Sheehan, Hrapchak, second edition - "Processing of Tissue", Chapter 3, p.83 important point number 9 in the Tissue-Handling Chart For Sectioning. Theory and Practice of Histological Techniques Bancroft, Gamblin, fifth edition - "Tissue Processing and Microtomy", Chapter 6, p.97 last sentence under the subheading floating out sections. lge ------------------------------ Message: 15 Date: Thu, 29 Apr 2004 21:14:05 EDT From: KAELPERS@aol.com Subject: Re: [Histonet] slide stainers - vendors To: Sue.Kapoor@uhsi.org, histonet@pathology.swmed.edu Message-ID: <6b.283d2342.2dc3025d@aol.com> Content-Type: text/plain; charset="US-ASCII" Hacker Linear Stainers...you can load until your heart is content. The only difficulty may be matching your current stain schedule to one that will be able to work on the hacker system. There is a timer you set permanently on the instrument and each rack will stay in that solution for that set time then prompt the instrument to lift and move to the next solution container. It is a great system, the only other factor is that it is a relatively long instrument and requires some space. lge ------------------------------ Message: 16 Date: Thu, 29 Apr 2004 22:11:00 -0400 From: "yan gao" Subject: [Histonet] Tissue array To: histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain Hi, Histonet. I am interested to get a few tissue array for different human cancers and cell array from human tumor cell lines. Any recommendation? Yan Gao Ph.D Norvatis _________________________________________________________________ [1]Stop worrying about overloading your inbox - get MSN Hotmail Extra Storage! References 1. http://g.msn.com/8HMBENUS/2737??PS= ------------------------------ Message: 17 Date: Fri, 30 Apr 2004 08:48:42 +0100 From: "Kemlo" Subject: RE: [Histonet] quickest freezing method AND maintain morphology of mouse brains for LCM then microarray??? To: "Karl Brand" , histonet@lists.utsouthwestern.edu Message-ID: <40800C1000030126@mk-cpfrontend-4.mail.uk.tiscali.com> Content-Type: text/plain; charset="iso-8859-1" London on a January day. Quickest way to freeze anyone's brain; luckily these Southeners are immune! __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM ------------------------------ Message: 18 Date: Fri, 30 Apr 2004 10:14:30 +0000 From: "Thom Jensen" Subject: RE: [Histonet] nail sectioning To: Histonet@lists.utsouthwestern.edu, mary.bliss@northwestpathology.com Message-ID: Content-Type: text/plain Have you tried 15% ammonia water. We have had success at softing the nail before processing and after if needed for sectioning. I have used this technique for horse hoff as well. Thom >From: "Bliss, Mary E." >To: >Subject: [Histonet] nail sectioning >Date: Thu, 29 Apr 2004 15:07:08 -0700 > >Hi to All, > > > >Does anyone have a good procedure for fixing and cutting in good >fingernail and toenail sections? We are trying to come up with >something which will allow us to produce a good stainable slide with ALL >of the tissue still intact on it! I have heard about using >trichloroacetic acid but can't seem to find a procedure anywhere. Have >also heard about /tried Nair and soapy water. Does anyone have any >information to share? > > > >Humbly yours, > > > >Mary E. Bliss > >Lead Histologist > >Northwest Pathology, P.S. > >3614 Meridian St. Suite 100 > >Bellingham, WA 98225 > >(360)734-2800 x601 > >(360)734-3818 FAX > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ [1]Stop worrying about overloading your inbox - get MSN Hotmail Extra Storage! References 1. http://g.msn.com/8HMBENUS/2737??PS= ------------------------------ Message: 19 Date: Fri, 30 Apr 2004 08:48:20 -0400 From: mark.lewis@thermo.com Subject: Re: [Histonet] nail sectioning To: "Bliss, Mary E." Cc: histonet-bounces@lists.utsouthwestern.edu, histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; charset=us-ascii Mary, Have you ever tried cutting Frozen sections of the unfixed toenail and fingernail ? You may be surprised to find they cut quite easily as a frozen section compared to a paraffin processed block. 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 "Bliss, Mary E." .com> cc: Sent by: Subject: [Histonet] nail sectioning histonet-bounces@lists.utsouth western.edu 04/29/2004 06:07 PM Hi to All, Does anyone have a good procedure for fixing and cutting in good fingernail and toenail sections? We are trying to come up with something which will allow us to produce a good stainable slide with ALL of the tissue still intact on it! I have heard about using trichloroacetic acid but can't seem to find a procedure anywhere. Have also heard about /tried Nair and soapy water. Does anyone have any information to share? Humbly yours, Mary E. Bliss Lead Histologist Northwest Pathology, P.S. 3614 Meridian St. Suite 100 Bellingham, WA 98225 (360)734-2800 x601 (360)734-3818 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 20 Date: Fri, 30 Apr 2004 08:45:50 -0500 From: "GUTIERREZ, JUAN" Subject: RE: [Histonet] laminin To: "Megan Kear" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Does it have to be polyclonal? We use DAKO's monoclonal with very good results. (1:20, no pretreatment). They also have a polyclonal, but I have not tried it. Good luck, Juan -----Original Message----- From: Megan Kear [mailto:Megan.Kear@hunter.health.nsw.gov.au] Sent: Wednesday, April 28, 2004 11:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] laminin Hi Histonetters Anybody using a good laminin polyclonal antibody. To be used on formalin fixed paraffin fixed embedded sections. thank you for your help. Megan Clarke HAPS IHC dept Newcastle Australia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 21 Date: Fri, 30 Apr 2004 09:22:17 -0500 From: "Nava, Josefa" Subject: [Histonet] Looking for ZAP-70and CD38 antibodies To: "'HISTONET@PATHOLOGY.SWMED.EDU'" Message-ID: <7A5E02BB0E0A2E409F971D889B200C298A53DB@phdex05.txhealth.org> Content-Type: text/plain; charset="iso-8859-1" Hello to everyone! Can someone tell me where I can get ZAP -70 and CD38 antibodies tnat will work well with paraffin sections and also with Ventana Benchmark/ XT stainer. I thank you very much. Josie 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. ------------------------------ Message: 22 Date: Fri, 30 Apr 2004 10:28:35 -0400 From: "Rebecca Barnhart" Subject: [Histonet] Container To: Message-ID: Content-Type: text/plain; charset=US-ASCII Our OR has asked that we get different containers for the larger specimens. Our current container is the basic container with a snap lid, they would like a screw top lid. The concern is if the container is dropped it will leak with the containers we have now. The only problem I am having is trying to find a gallon screw top container. Any ideas? Again thank you in advance. Becky ------------------------------ Message: 23 Date: Fri, 30 Apr 2004 10:44:50 -0400 From: "Gladney, Diane C Ms MACH" Subject: RE: [Histonet] Container To: 'Rebecca Barnhart' , histonet@lists.utsouthwestern.edu Message-ID: <9D41AB7C56F8304F98537ABD87B249F66261EF@DASMTHGBZ001> Content-Type: text/plain; charset="iso-8859-1" You may have a hard time finding gallon size specimen containers with screw tops. Our OR has a large deep tray with a handle that they use to transport specimens to the Histology Dept. The containers that they are now using have lids that snap on snuggly and I sometimes have difficulty getting these lids off. How do they transport the containers to your lab? I have found that screw top containers will leak like crazy if the lids are not put on straight. We have trouble from our clinics even with the small screw top containers leaking because they won't take the time to put the lids on straight. Diane Diane C. Gladney, HT (ASCP) Histology /Cytology Supervisor Moncrief Army Community Hospital P.O. BOX 484 4500 Stuart Ave. FT. Jackson, SC 29207 (803) 751-2530 DSN 734-2530 EMAIL: diane.gladney@se.amedd.army.mil OR dcgx1@aol.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rebecca Barnhart Sent: Friday, April 30, 2004 10:29 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Container Our OR has asked that we get different containers for the larger specimens. Our current container is the basic container with a snap lid, they would like a screw top lid. The concern is if the container is dropped it will leak with the containers we have now. The only problem I am having is trying to find a gallon screw top container. Any ideas? Again thank you in advance. Becky _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 24 Date: Fri, 30 Apr 2004 07:49:24 -0700 From: "Kari Bradshaw" Subject: RE: [Histonet] Job interview questions To: "Richard Larson" , Message-ID: Content-Type: text/plain; charset="us-ascii" Dear Histonetters, Here is a list that I have developed over the last few years. I hope it's helpful. Kari Bradshaw, HT(ASCP) Laboratory Manager Lower Columbia Pathologists 1217 14th Ave Longview, WA 98632 (360)425-5620 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Richard Larson Sent: Thursday, April 29, 2004 8:03 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Job interview questions I have a job interview next week with a private lab as a histotech. I have my ht certification plus four years of experience doing embedding, staining and some microtomy. I have been out of the lab the past three years doing other work but would now like to return to the histology profession. If anyone has had a recent interview, I would be interested in learning what sort of questions interviewers will likely ask, so that I can be more prepared. Thanks for any replies. --------------------------------- Do you Yahoo!? Win a $20,000 Career Makeover at Yahoo! HotJobs _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 25 Date: Fri, 30 Apr 2004 11:11:03 -0400 From: "White, Lori" Subject: [Histonet] Contamination of processor alcohols To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain Hi, We use an alcohol/xylene recycler and routinely test the alcohol for xylene contamination before recycling. We have noticed a significant proportion of our alcohols are contaminated when tested. Through process of elimination, we have determined that our contamination is coming from the alcohols on the processor. We replaced all of our alcohols with clean, non-recycled alcohol and ran a cycle. The alcohols were tested the following morning and one of the alcohols was contaminated. Has anyone had experience with this? Thanks, Lori Lakeridge Health Corporation Ontario, Canada ------------------------------ Message: 26 Date: Fri, 30 Apr 2004 11:26:09 -0400 From: "Greer, Patricia" Subject: RE: [Histonet] Container To: "Rebecca Barnhart" , Message-ID: Content-Type: text/plain; charset="us-ascii" Becky, Nalgene makes a 1 gallon wide-mouth container with screw top lid - we often get specimens sent to us in these containers. I'm sure any of the supply catalogs would have them. Actually I just looked at the Fisher catalog and they have several sizes of wide mouth bottles listed. You can go to their web site (www.fishersci.com) and search for bottles in their general laboratory section. Pat Greer Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road Atlanta, GA 30333 Our OR has asked that we get different containers for the larger specimens. Our current container is the basic container with a snap lid, they would like a screw top lid. The concern is if the container is dropped it will leak with the containers we have now. The only problem I am having is trying to find a gallon screw top container. Any ideas? Again thank you in advance. Becky _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 27 Date: Fri, 30 Apr 2004 11:49:33 -0400 From: "Smith, Allen" Subject: RE: [Histonet] Zinc formalin and 10% NBF To: Cc: histonet@pathology.swmed.edu Message-ID: <494304423C63E246A5CF87A3AEEB577011B5E7@bumail01.barrynet.barry.edu> Content-Type: text/plain; charset="iso-8859-1" It's not the periodic table, but the electromotive series. Ions of elements at the top of the series will replace free elements (only on the surface of any but the tiniest particles) lower in the series. The ion is reduced to the free element and the lower down free element is oxidized to the ion. Zinc is pretty low in the electromotive series, so many ions can be reduced by zinc metal, converting the zinc metal to zinc ions. Unfortunately, the zinc in tissues is there as ions already. Any metal low enough in the series to reduce zinc ions to zinc metal would be such a good reducing agent that it would reduce water to hydrogen. 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 Kemlo Sent: Tuesday, April 27, 2004 2:43 AM To: jkiernan@uwo.ca; Wendy England Cc: histonet@pathology.swmed.edu Subject: Re: [Histonet] Zinc formalin and 10% NBF Can't you test the residual fixative for the presence or absence of zinc? My Chemistry is not what it was but can't you do something with an element and the double decomposition of a salt? Something to do with an element higher up or lower down the Periodic Table that displaces an element from a salt that is the other way round, can't remember. But NBF doesn't contain the heavy metal does it? Why is it important to know? All I can remember is that zinc formalin made the tissue brittle, or was that lead? The nuclear stain was rather deep too. Hope that helps. __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM _______________________________________________ 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: 28 Date: Fri, 30 Apr 2004 11:21:14 -0500 From: Don.Birgerson@leica-microsystems.com Subject: Re: [Histonet] thick sections in zebrafish To: "Eric Tytell" Cc: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Hi Eric, I have read your question with some curiosity. If the JB4 supports your specimen without distortion, why not use a motorized rotary microtome to section? If you have any questions, please feel free to phone me. Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 7:00 - 4:00 CDT "Eric Tytell" To: Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] thick sections in zebrafish western.edu 04/28/2004 02:40 PM Hi all -- I'm trying to get thick (between 50 and 100 micron) transverse sections of adult zebrafish (15 to 20mm long). I'm concerned about the 3D structure of the muscle tissue, so I'd like to have the specimens embedded in some supporting medium. I've had good luck using plastic (JB-4) sectioned with a saw, but the trouble with that method is that I lose 300microns to the saw each time I cut. I'd like to try embedding in a different medium, but I haven't been able to find a protocol. Has anyone tried to do something like this? I'm currently experimenting with paraffin embedding. Specifically, does anyone have suggestions for how long and in what to decalcify? I currently have both EDTA and Poly-NoCal available. Also, how long should I infiltrate in paraffin? Alternatively, I've tried using a cryostat microtome, but the muscle tissue seems to be damaged by the freezing or possibly by desiccation. Is it necessary/possible to decalcify for frozen sections? Any suggestions for avoiding freezing damage? Also, does anyone know of a good polyclonal antibody for zebrafish skeletal muscle? I'm not looking for anything too specific -- just enough to distinguish skeletal muscle from bone and collagen under low magnification confocal imaging. I'd appreciate any suggestions. Thanks in advance, Eric Tytell ------------------------------------------ Eric Tytell Museum of Comparative Zoology Laboratories Harvard University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 29 Date: Fri, 30 Apr 2004 09:44:16 -0700 (PDT) From: Scott Mordue Subject: Re: [Histonet] Tissue array To: histonet@pathology.swmed.edu Message-ID: <20040430164416.15176.qmail@web42002.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hi Yan Gao, I think you should contact Zymed. They have very high quality Tissue Microarray, especially the human ones. 800-874-4494 Scott CSU -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of yan gao Sent: Thursday, April 29, 2004 7:11 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Tissue array Hi, Histonet. I am interested to get a few tissue array for different human cancers and cell array from human tumor cell lines. Any recommendation? Yan Gao Ph.D Norvatis _________________________________________________________________ [1]Stop worrying about __________________________________ Do you Yahoo!? Win a $20,000 Career Makeover at Yahoo! HotJobs http://hotjobs.sweepstakes.yahoo.com/careermakeover ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 5, Issue 47 *************************************** From mcauliff <@t> umdnj.edu Mon May 3 11:46:52 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Testes Bouin's fluid In-Reply-To: References: Message-ID: <4096777C.40501@umdnj.edu> Dear Dr. Balaji: A few questions to clarify your problem: How long were the testes fixed in Bouin? Was fixation by immersion or perfusion? How long was the tissue washed in 70% ethanol? Why was the tissue 'refixed' in buffered formalin? Tissues properly fixed in Bouin should not need to be refixed. Exactly what is it about the morphology/staining that is "not that great"? Bouin's is an excellent fixative and, assuming it is made correctly and the tissues are fixed promptly and for sufficient time, the most likely source of your problems is not the fixative. Geoff balajimr@drreddys.com wrote: >Dear Histonetter, > >We have been preserving testes (of rats and mouse) in bouin's solution and >after 24 hrs washing them with 70% alcohol and fixing it back in 10%NBF. >But staining later with H & E is not that great. Can anybody suggest me >the best protocol for fixing,processing and staining of testes. > >Dr. Balaji >Dept. Pre clinical safety evaluation, >Discovery research, >Dr. Reddys Laboratories ltd. Bollaram Road, >Miyapur, Hyderabad, 500 049 > Andhra Pradseh, INDIA >_______________________________________________ >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 cjohnston <@t> mdanderson.org Mon May 3 09:05:34 2004 From: cjohnston <@t> mdanderson.org (cjohnston@mdanderson.org) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Guinea Pig Antibodies Message-ID: I am looking for antibodies that react with or are anti guinea pig. Specifically CD31 and Factor VIII. Thanks, Carol Carol M. Johnston HT(ASCP) M.D. Anderson Cancer Center 1515 Holcombe Blvd. Unit 443 Houston, Texas 77030 713-745-4625 From SCheasty <@t> ahs.llumc.edu Mon May 3 10:09:00 2004 From: SCheasty <@t> ahs.llumc.edu (Cheasty, Sandra) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Procedure for SAB & Retic Stains Message-ID: <2E50F33F91EEDA46A77BC3B2575BB0910587D7@mars.llumc.edu> Hello everyone, We are determining which stains we will do in our lab which receives mostly outpatient biopsies. We don't often do a Retic or Sulfated Alcian Blue 1 or 2 a year) and these are 2 that we are considering sending out. Does anyone have a recent procedure for these stains that may be less complicated that the AFIP or Sheehan Hrapchak versions? There are kits for Retic stain, but the only ones I've found have been very expensive ($500 and up). Your help and suggestions are much appreciated. Sandy Confidentiality Note: The preceding e-mail message (including any attachments) contains information that may be confidential, protected by applicable legal privileges, or constitute non-public information. It is intended to be conveyed only to the designated recipient(s). If you are not an intended recipient of this message, please notify the sender by replying to this message and then delete it from your system. Use, dissemination, distribution or reproduction of this message by unintended recipients is not authorized and may be unlawful. From jkiernan <@t> uwo.ca Mon May 3 09:40:29 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Testes Bouin's fluid References: Message-ID: <409659DD.C581668C@uwo.ca> It does not make sense to put specimens in neutral buffered formaldehyde after fixing them in Bouin's fluid. Bouin's is, for many purposes, a better fixative than NBF because it confers characteristic patterns of chromatin in nuclei of different cell types and it also permits brighter staining of collagen and cytoplasm with anionic dyes. Bouin's fluid extracts cytoplasmic RNA and it can also make DNA Schiff-positive (Feulgen hydrolysis), and can damage red blood cells. Staining by both haemalum and eosin is stronger after Bouin's than after NBF, and in many tissues the microanatomy, as seen in paraffin sections, is more lifelike. There is less differential shrinkage of cells, tubules etc than you see in paraffin sections of specimens fixed in neutral formaldehyde. -- ------------------------------- 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 _______________________________ balajimr@drreddys.com wrote: > > Dear Histonetter, > > We have been preserving testes (of rats and mouse) in bouin's solution and > after 24 hrs washing them with 70% alcohol and fixing it back in 10%NBF. > But staining later with H & E is not that great. Can anybody suggest me > the best protocol for fixing,processing and staining of testes. > > Dr. Balaji > Dept. Pre clinical safety evaluation, > Discovery research, > Dr. Reddys Laboratories ltd. Bollaram Road, > Miyapur, Hyderabad, 500 049 > Andhra Pradseh, INDIA > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From csquire <@t> tds.net Mon May 3 11:31:26 2004 From: csquire <@t> tds.net (Craig Squire) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Testes Bouin's fluid References: <409659DD.C581668C@uwo.ca> Message-ID: <002b01c4312c$16112180$6701a8c0@DESK> I work in a contract research lab and we keep almost all of our testes, and sometimes epididymides, in 70% alcohol permanently. We rinse with distilled water and then store in 70%. All of the slides are then stained with H&E and they turn out really nice. When we place the tissue in 70% COH we do not trim the tissue right away, we wait a few days. Hope this helps. Heather Squire ----- Original Message ----- From: "John Kiernan" To: Cc: Sent: Monday, May 03, 2004 9:40 AM Subject: Re: [Histonet] Testes Bouin's fluid > It does not make sense to put specimens in neutral > buffered formaldehyde after fixing them in Bouin's > fluid. Bouin's is, for many purposes, a better > fixative than NBF because it confers characteristic > patterns of chromatin in nuclei of different cell > types and it also permits brighter staining of > collagen and cytoplasm with anionic dyes. Bouin's > fluid extracts cytoplasmic RNA and it can also > make DNA Schiff-positive (Feulgen hydrolysis), and > can damage red blood cells. > > Staining by both haemalum and eosin is stronger > after Bouin's than after NBF, and in many tissues > the microanatomy, as seen in paraffin sections, is > more lifelike. There is less differential shrinkage > of cells, tubules etc than you see in paraffin > sections of specimens fixed in neutral formaldehyde. > -- > ------------------------------- > 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 > _______________________________ > balajimr@drreddys.com wrote: > > > > Dear Histonetter, > > > > We have been preserving testes (of rats and mouse) in bouin's solution and > > after 24 hrs washing them with 70% alcohol and fixing it back in 10%NBF. > > But staining later with H & E is not that great. Can anybody suggest me > > the best protocol for fixing,processing and staining of testes. > > > > Dr. Balaji > > Dept. Pre clinical safety evaluation, > > Discovery research, > > Dr. Reddys Laboratories ltd. Bollaram Road, > > Miyapur, Hyderabad, 500 049 > > Andhra Pradseh, INDIA > > _______________________________________________ > > 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 Barry.R.Rittman <@t> uth.tmc.edu Mon May 3 11:59:38 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Testes Bouin's fluid Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0635A4A@UTHEVS3.mail.uthouston.edu> An important point here is that during fixation in Bouin's some of the protein picrates that are formed are soluble in water but are rendered insoluble when the tissue is placed in 70% or higher percentage alcohol. Placing tissue in water (and therefore presumably in aqueous formalin) allows some of the picrates to leach out of the tissue. I like Bouin's for fixing tissues such as testes (animal testes that is - we don't get too many from patients here in the dental school!!) as the coarse precipitate allows nuclear details to be clearer. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Craig Squire Sent: Monday, May 03, 2004 11:31 AM To: balajimr@drreddys.com Cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Testes Bouin's fluid I work in a contract research lab and we keep almost all of our testes, and sometimes epididymides, in 70% alcohol permanently. We rinse with distilled water and then store in 70%. All of the slides are then stained with H&E and they turn out really nice. When we place the tissue in 70% COH we do not trim the tissue right away, we wait a few days. Hope this helps. Heather Squire ----- Original Message ----- From: "John Kiernan" To: Cc: Sent: Monday, May 03, 2004 9:40 AM Subject: Re: [Histonet] Testes Bouin's fluid > It does not make sense to put specimens in neutral > buffered formaldehyde after fixing them in Bouin's > fluid. Bouin's is, for many purposes, a better > fixative than NBF because it confers characteristic > patterns of chromatin in nuclei of different cell > types and it also permits brighter staining of > collagen and cytoplasm with anionic dyes. Bouin's > fluid extracts cytoplasmic RNA and it can also > make DNA Schiff-positive (Feulgen hydrolysis), and > can damage red blood cells. > > Staining by both haemalum and eosin is stronger > after Bouin's than after NBF, and in many tissues > the microanatomy, as seen in paraffin sections, is > more lifelike. There is less differential shrinkage > of cells, tubules etc than you see in paraffin > sections of specimens fixed in neutral formaldehyde. > -- > ------------------------------- > 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 > _______________________________ > balajimr@drreddys.com wrote: > > > > Dear Histonetter, > > > > We have been preserving testes (of rats and mouse) in bouin's > > solution and > > after 24 hrs washing them with 70% alcohol and fixing it back in > > 10%NBF. But staining later with H & E is not that great. Can anybody > > suggest me the best protocol for fixing,processing and staining of > > testes. > > > > Dr. Balaji > > Dept. Pre clinical safety evaluation, > > Discovery research, > > Dr. Reddys Laboratories ltd. Bollaram Road, > > Miyapur, Hyderabad, 500 049 > > Andhra Pradseh, INDIA > > _______________________________________________ > > 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 nmuvarak <@t> facstaff.wisc.edu Mon May 3 11:18:41 2004 From: nmuvarak <@t> facstaff.wisc.edu (NIDAL E MUVARAK) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Antibody for Collagen Type I in Mice Message-ID: <54c21b549a4a.549a4a54c21b@wiscmail.wisc.edu> Dear Histonetters, Can anyone recommend an antibody/vendor (polyclonal or monoclonal) for collagen type I. I'm interested in staining mouse lungs and pulmonary arteries. I used frozen sections embedded in OCT and fixed in aceton after sectioning. Your help is much appreciated. Thanks. Regareds, Nidal E Muvarak Associate Research Specialist Vascular Tissue Biomechanics Laboratory Department of Biomedical Engineering University of Wisconsin-Madison 1550 Engineering Dr.; Rm. 2158 Madison, WI 53706-1609 Lab: (608) 265-8921; Office: (608) 265-4205; Home: (608) 256-7934; Cell: (608) 332-6068 http://vtb.bme.wisc.edu From Sue.Kapoor <@t> uhsi.org Mon May 3 13:10:19 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Hacker Instruments Microwave Tissue Processor Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F55C9@khmcexch.uhsi.org> Hello histoland, does anyone use Hacker Instruments microwave tissue processor, either model...RHS-1 or RHS-2? If so, can you tell me how you like it? Thank you, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From clarkda <@t> ohsu.edu Mon May 3 08:39:17 2004 From: clarkda <@t> ohsu.edu (David Clark) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] cutting rat brains Message-ID: Hello, I was wandering if any of you wonderful Histonet people who have much experience cutting rat brains can offer some assistance in trying to cut good quality sections. We either encounter lots of wrinkles or tissue seperation from the paraffin sections once placed on the waterbath. Is this a processing problem or do you think it is strictly a sectioning problem? Any suggestions would be appreciated. Thanks, David Clark Neuropathology OHSU From marilyn.j.easton <@t> gsk.com Mon May 3 14:39:09 2004 From: marilyn.j.easton <@t> gsk.com (marilyn.j.easton@gsk.com) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] cutting rat brains Message-ID: Sounds like a problem with your water bath temperature. Too many wrinkles means your water bath is too cold and quick separation is an indication that your water bath is too hot or you have a processing problem. We generally use a 38 to 42 degree centigrade water bath temperature depending on how quickly you can pick up your section. The rat cerebellum tends to separate more quickly than the cerebrum which is why we block the tissue so that the cerebellum is picked up first on the slide and the cerebrum can be "dangled" in the waterbath a few seconds longer if needed. Processing plays a big part in how well your brain tissue sections, so you should try picking up your sections at different temperatures to get a feel for what works best with your tissue. Marilyn Easton, B.A., HT/HTL (ASCP), LAT (AALAS) Scientist, Pathology Dept., Safety Assessment GlaxoSmithKline 919-483-7891 "David Clark" Sent by: To: Histonet@lists.utsouthwestern.edu histonet-bounces@lists.utsouth western.edu cc: Subject: [Histonet] cutting rat brains 03-May-2004 09:39 Hello, I was wandering if any of you wonderful Histonet people who have much experience cutting rat brains can offer some assistance in trying to cut good quality sections. We either encounter lots of wrinkles or tissue seperation from the paraffin sections once placed on the waterbath. Is this a processing problem or do you think it is strictly a sectioning problem? Any suggestions would be appreciated. Thanks, David Clark Neuropathology OHSU _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Tom_Wells <@t> bcit.ca Mon May 3 17:52:58 2004 From: Tom_Wells <@t> bcit.ca (Tom Wells) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Laboratory design Message-ID: I am faced with the task of re-designing an existing teaching laboratory with the goal being improved ventilation and safety in mind. This of course has to be done on a limited educational budget. This laboratory currently houses roughly 20 students at a time. We have a tissue processor and an automated stainer in one area. A small fumehood which I currently use for manual coverslipping. We have several small benches which are used for grossing and finally 6 large benches with equipment for approximately 3 students per bench (microtome, waterbath, sink and staining tray for each student). Does anyone have any suggestions on how I can approach this project? Thanks. Tom From rnishi <@t> uci.edu Mon May 3 20:28:23 2004 From: rnishi <@t> uci.edu (Rebecca Nishi) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Histobath or alternative? Message-ID: Does anyone currently use the histobath with isopentane, or a similar bath? Currently we freeze rodent brain and spinal cords in isopentane cooled to -65 (in a plastic beaker surrounded by dry ice). We would like to purchase a bath to get rid of the need for dry ice, and to more easily control the temperature. I know that Thermo Electron (Shandon) currently makes a histobath, but I understand that the temperature is not controllable? Is this true? Does anyone know of any alternatives. We just need something for a benchtop, nothing too large, and probably under 5K. Thanks, Rebecca Nishi UC Irvine From mlm11 <@t> cornell.edu Tue May 4 07:28:59 2004 From: mlm11 <@t> cornell.edu (Mary Lou Norman) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Testes Bouin's fluid Message-ID: <5.2.1.1.2.20040504081734.00b52ae8@postoffice9.mail.cornell.edu> These tissues are small enough that they don't need to be fixed for 24 hours. Try 8 to 12 hours, skip the NBF, wash in water for a bit, and rinse with several changes of 70% ETOH to remove picric acid. More acid will leach out during processing. Provided the tissues are no more than 3mm thick, 2-3 hours in 80% ETOH and 1 hour each for the rest of your processing should give you decent sections that will stain adequately with any H&E. If you don't leach out enough picric acid, your sections will compress when cutting, so use ice. Just another way to do it. Mary Lou Norman College of Veterinary Medicine Cornell From carl.hobbs <@t> kcl.ac.uk Tue May 4 02:29:44 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] re ssDNA Message-ID: <001c01c431a9$92a86500$e8345c9f@Carlos> I tried Chemicon's MAB3299 witout success. Both their "special detection protocol" and heat- induced AR). If you try it be interested in your comments. I suppose an anti activated- Caspase 3 is the best way at the moment, to detect apoptotic cells? From carl.hobbs <@t> kcl.ac.uk Mon May 3 13:01:22 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] re mounting media for NBT/BCIP Message-ID: <002001c43138$a4fab580$c6949a51@home> Why an aq. one? Thoroughly air-dry, soak in xylene for 10mins, then mount in DPX? Best resolution. Aq. mountants don't give as good a resolution, partic. when preparing photomicrographs/using high power. Best aq. mountants for me: gelatin-based ones eg Gelvatol, that set at RT, or Mowiol-based mountant that set, too( yes, it is designed for fluorescent dyes) --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.668 / Virus Database: 430 - Release Date: 24/04/2004 From siksik03 <@t> comcast.net Tue May 4 08:20:26 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Histobath or alternative? In-Reply-To: References: Message-ID: Rebecca You need the Hacker/Bright Clini-RF. The temperature can be accurately controlled, and you won't need dry ice. I have no idea what it costs, but it works like a charm. best regards, Steven Slap At 6:28 PM -0700 5/3/04, Rebecca Nishi wrote: >Does anyone currently use the histobath with isopentane, or a similar bath? > >Currently we freeze rodent brain and spinal cords in isopentane cooled to >-65 (in a plastic beaker surrounded by dry ice). We would like to purchase a >bath to get rid of the need for dry ice, and to more easily control the >temperature. > >I know that Thermo Electron (Shandon) currently makes a histobath, but I >understand that the temperature is not controllable? Is this true? Does >anyone know of any alternatives. We just need something for a benchtop, >nothing too large, and probably under 5K. From fabienfuente <@t> yahoo.fr Tue May 4 08:48:49 2004 From: fabienfuente <@t> yahoo.fr (Fabien FUENTE) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Mac Weals tetrachrome Message-ID: Hi all, Does someone has a protocol for Mac Weals tetrachrome? This stain is supposed to be good in bone staining. Is there a difference in staining quality between it and RBS (Rapid Bone Stain)? Any help will be useful. Regards Fabien Fuente From Stephen.Eyres <@t> sanofi-synthelabo.com Tue May 4 10:11:12 2004 From: Stephen.Eyres <@t> sanofi-synthelabo.com (Stephen.Eyres@sanofi-synthelabo.com) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] cutting rat brains Message-ID: Hi David, Creases come from waterbath temp being too low. Tissue separation can be caused be water temp being too high and/or you have left the block on water too long. Cheers Steve "David Clark" To: Histonet@lists.utsouthwestern.edu Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] cutting rat brains western.edu 03/05/2004 14:39 Hello, I was wandering if any of you wonderful Histonet people who have much experience cutting rat brains can offer some assistance in trying to cut good quality sections. We either encounter lots of wrinkles or tissue seperation from the paraffin sections once placed on the waterbath. Is this a processing problem or do you think it is strictly a sectioning problem? Any suggestions would be appreciated. Thanks, David Clark Neuropathology OHSU _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rnishi <@t> uci.edu Tue May 4 10:19:04 2004 From: rnishi <@t> uci.edu (Rebecca Nishi) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Mouse and rat injured spinal cord - polyester wax Message-ID: Has anyone used polyester wax embedding for mouse or rat injured spinal cord. We would like to use a low melt wax instead of paraffin or plastic to preserve some of the antigens we are interested in, but we also are interested in doing stereology, so we want consistent and a complete set of sections. Also, the injured area poses many problems for sectioning. Does anyone have any tips for cutting and staining with this wax? I am going to use a rotary microtome. Thanks Rebecca From rnishi <@t> uci.edu Tue May 4 10:27:29 2004 From: rnishi <@t> uci.edu (Rebecca Nishi) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Histobath or alternative? In-Reply-To: Message-ID: Thanks Steven, I am trying to find this on hackerinstruments.com. Is this the correct website? Maybe it is just down today. Thanks, Rebecca On 5/4/04 6:20 AM, "Steven E. Slap" wrote: > Rebecca > > You need the Hacker/Bright Clini-RF. The temperature can be > accurately controlled, and you won't need dry ice. I have no idea > what it costs, but it works like a charm. > > best regards, > Steven Slap > > At 6:28 PM -0700 5/3/04, Rebecca Nishi wrote: >> Does anyone currently use the histobath with isopentane, or a similar bath? >> >> Currently we freeze rodent brain and spinal cords in isopentane cooled to >> -65 (in a plastic beaker surrounded by dry ice). We would like to purchase a >> bath to get rid of the need for dry ice, and to more easily control the >> temperature. >> >> I know that Thermo Electron (Shandon) currently makes a histobath, but I >> understand that the temperature is not controllable? Is this true? Does >> anyone know of any alternatives. We just need something for a benchtop, >> nothing too large, and probably under 5K. > From sharon.willman <@t> bms.com Tue May 4 10:28:19 2004 From: sharon.willman <@t> bms.com (Sharon E Willman) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] S-100 background staining in rat tissue Message-ID: <4097B693.B5A75FFE@bms.com> Hi, We are currently developing S-100 immunostain in rat tissue on the Ventanna. We have used several protocols, but seems to have a lot of background staining. Does anyone have any suggestions as to how to decrease the background? We have decreased the time for the primary, added AB blocker to the protocol, but still have a great deal of background staining. Any help would be most appreciated. Thank you in advance! Sharon Willman From la.sebree <@t> hosp.wisc.edu Tue May 4 10:27:42 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Looking for Amy from PathLogic Message-ID: Amy, Please call me. I've been unsuccessful in faxing our C4d protocol to you. 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 Stephen.Eyres <@t> sanofi-synthelabo.com Tue May 4 10:44:42 2004 From: Stephen.Eyres <@t> sanofi-synthelabo.com (Stephen.Eyres@sanofi-synthelabo.com) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Mouse and rat injured spinal cord - polyester wax Message-ID: Hi Rebecca, Many, many years ago I used 37 degree C MPT polyester wax. There are several major problems using the stuff. Key one is temperature. Blocks and knife were kept in the fridge, with both brought out just prior to use -we used a Cambridge rocker which meant transfer of knife to and from the fridge was easy. In the summer in our lab which had no air conditioning, there are minutes of cutting time available before the block and kife warms up and sectioning has to stop. One paper I read partly overcame this narrow curring window by using a flower sieve packed with dry ice suspended above the microtome. our resources did not stretch that far in those days (mid 70's). Once you have your sections, we had to use a starch based adhesive which was added to the slide and sections floated directly onto the pool of adesive. The trick was to ensure one end of the ribbon overlapped the end of the slide so that after the short period of time the sections took to expand and de-crease, the slide could be tipped allowing the adhesive to run away leaving a nice ribbon on the slide. The rest was easy!! Having read all of this, it comes as no surprise that my advice is to try every other option for getting what you want first. Best of luck. Steve Rebecca Nishi Sent by: To: histonet-bounces@lists.utsouth cc: western.edu Subject: [Histonet] Mouse and rat injured spinal cord - polyester wax 04/05/2004 16:19 Has anyone used polyester wax embedding for mouse or rat injured spinal cord. We would like to use a low melt wax instead of paraffin or plastic to preserve some of the antigens we are interested in, but we also are interested in doing stereology, so we want consistent and a complete set of sections. Also, the injured area poses many problems for sectioning. Does anyone have any tips for cutting and staining with this wax? I am going to use a rotary microtome. Thanks Rebecca _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Tue May 4 10:49:20 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Charging IHC Message-ID: It is my understanding that antibodies labeled as "for in vitro diagnostic use" or "ASR" can be billed to patients; antibodies labeled "for research use only" cannot. Is that correct? Thank you. Richard Cartun From CRIST123 <@t> aol.com Tue May 4 10:59:42 2004 From: CRIST123 <@t> aol.com (CRIST123@aol.com) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] ISH LAMBDA PROBLEM HELP! Message-ID: <3E318A99.5E70E30F.0067E063@aol.com> Our hospital has began to run K&L Ish using Benchmark/ventanna. WE are having problems with overstaining of plasma cells on the clot section of Lambda only. The Kappa looks fine on clot section. Any ideas on what problem can be? Our tonsil control looks great, only patient clot section. We are thinking it may be the preparation of clot section prior to staining (possibly the anticoagulant used). Please write if you have had the same problem and have resolved this. Thanks, Cris and Angie, Good Sam Hospital, Cincinnati From laurie.colbert <@t> huntingtonhospital.com Tue May 4 11:00:59 2004 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Histobath or alternative? Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5BED7@EXCHANGE1.huntingtonhospital.com> We have the Thermo Electron (Shandon) histobath. We use it to freeze frozen sections and it works well for us. I don't know how it would work for your applications. I'm assuming the brains and spinal cords are fairly small, so it would probably be ok. You cannot control the temp, but that does not affect what we do. Good luck. Laurie Colbert -----Original Message----- From: Rebecca Nishi [mailto:rnishi@uci.edu] Sent: Monday, May 03, 2004 6:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histobath or alternative? Does anyone currently use the histobath with isopentane, or a similar bath? Currently we freeze rodent brain and spinal cords in isopentane cooled to -65 (in a plastic beaker surrounded by dry ice). We would like to purchase a bath to get rid of the need for dry ice, and to more easily control the temperature. I know that Thermo Electron (Shandon) currently makes a histobath, but I understand that the temperature is not controllable? Is this true? Does anyone know of any alternatives. We just need something for a benchtop, nothing too large, and probably under 5K. Thanks, Rebecca Nishi UC Irvine _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kurnelmi <@t> umdnj.edu Tue May 4 11:33:33 2004 From: kurnelmi <@t> umdnj.edu (Michael Kurnellas) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Luxol Fast blue Message-ID: <1083688413.4097c5dd102f9@webmail.umdnj.edu> I was told that Luxol fast blue staining should be done on paraffin embedded tissue. Can I also do this stain on sucrose-embedded tissue, frozen, and then placed in OCT compound before 10um sections were taken? Thanks. Mike K. From cwscouten <@t> myneurolab.com Tue May 4 11:54:24 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Histobath or alternative? Message-ID: Temperature control above -80 C is an extra price option on the Cold Snap unit. 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 Laurie Colbert Sent: Tuesday, May 04, 2004 11:01 AM To: Rebecca Nishi; Histonet (E-mail) Subject: RE: [Histonet] Histobath or alternative? We have the Thermo Electron (Shandon) histobath. We use it to freeze frozen sections and it works well for us. I don't know how it would work for your applications. I'm assuming the brains and spinal cords are fairly small, so it would probably be ok. You cannot control the temp, but that does not affect what we do. Good luck. Laurie Colbert -----Original Message----- From: Rebecca Nishi [mailto:rnishi@uci.edu] Sent: Monday, May 03, 2004 6:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histobath or alternative? Does anyone currently use the histobath with isopentane, or a similar bath? Currently we freeze rodent brain and spinal cords in isopentane cooled to -65 (in a plastic beaker surrounded by dry ice). We would like to purchase a bath to get rid of the need for dry ice, and to more easily control the temperature. I know that Thermo Electron (Shandon) currently makes a histobath, but I understand that the temperature is not controllable? Is this true? Does anyone know of any alternatives. We just need something for a benchtop, nothing too large, and probably under 5K. Thanks, Rebecca Nishi UC Irvine _______________________________________________ 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 ramona.tolliver <@t> yale.edu Tue May 4 12:42:24 2004 From: ramona.tolliver <@t> yale.edu (Ramona Tolliver) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Histology Manager Opening at Yale University Message-ID: <6.0.1.1.2.20040504133359.01e99a10@email.med.yale.edu> Good afternoon, Yale University has a Histology Manager (1st shift) position available from 7:00 a.m. - 3:00 p.m. If you know of anyone who may be interested in an outstanding growth opportunity in a department committed to excellence in patient care, teaching, and discovery, please forward this information on to them. Salary is commensurate with experience. Relocation assistance is available. The position is posted online at : http://websrv.its.yale.edu/hr/cgi-bin/jobpost.plx?numreq=1&type=frags&extern=1&page=mp+2&keywords=MSGN12188&summary=yes&datesum=yes&deptsum=yes&salsum=yes&reqsum=yes Inquiries should be sent to ramona.tolliver@yale.edu. Thank you for your time! Ramona E. Tolliver *If you have difficulty with the link provided above, please go to www.yale.edu/jobs and search Managerial and Professional source code 12188. The position is listed as Manager III. Ramona E. Tolliver Human Resource Manager Yale University School of Medicine Department of Pathology 310 Cedar Street Lauder Hall 108 P.O. Box 208023 New Haven, CT 06520-8023 Office: (203) 785-6689 Fax: (203) 782-9471 From gcallis <@t> montana.edu Tue May 4 12:44:33 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] ATTN: Fabian on MacNeals tetrachrome versus RBS Message-ID: <3.0.6.32.20040504114433.00c0a778@gemini.msu.montana.edu> What you are looking for is MacNeals Tetrachrome. I posted the recipe on Histonet in the last month so go back in Histonet archives for information. Also, are you trying to do thick ground/polished bone sections embedded in methylmethacrylate? or microtomed PMMA sections? We always 1% formic acid etched bone sections for 30 sec to 1 min, then rinsed in tap water well, blotted dry before staining. This results in better bone components staining, if you want to see Haversian systems, caniliculi, old and new osteons, etc. If you do not acid etch with RBS, all you see is red counterstained bone, and blue to blue green soft tissue components and nuclei. RBS or Sandersons rapid bone stain is entirely different than MacNeals, tintorial differences are apparent after you use these solutions. One can also combine a toludine blue staining method (Sterchi and Eurell) with MacNeals and have a brilliantly stained bone section. RBS is a potassim permanganate oxidized methylene blue that results in production of various other dye components due to the oxidation of the methylene blue molecule (toluidine blue, azure A, azure B, thionin are some of the components). MacNeals is excellent, in fact, I think far superior for demonstration of osteoid as compared to RBS. You can purchase RBS from Surgipath or you can make up Stevenels blue, these are the same stain solutions, although made up differently. I would strongly advise buying RBS, as making Stevenels blue is a long, tedious and messy process. You should make up the MacNeals in house as we found commercial preparations of this stain to be very poor for bone work. How you use the stain depends on how you prepare the section. MacNeals is stable for years, at least the inhouse solution. Hi all, Does someone has a protocol for Mac Weals tetrachrome? This stain is supposed to be good in bone staining. Is there a difference in staining quality between it and RBS (Rapid Bone Stain)? Any help will be useful. Regards Fabien Fuente 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 DonnaWillis <@t> texashealth.org Tue May 4 13:05:20 2004 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Charging IHC Message-ID: <5C6CBCCEB04B894C8BD15B312487F7B201A330BA@ftwex01.txhealth.org> Richard, That is the way we charge. We have asked the antibody manufacture to change the product sheets from RUO to ASR but it is very slow comming. Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Richard Cartun Sent: Tuesday, May 04, 2004 10:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Charging IHC It is my understanding that antibodies labeled as "for in vitro diagnostic use" or "ASR" can be billed to patients; antibodies labeled "for research use only" cannot. Is that correct? Thank you. Richard Cartun _______________________________________________ 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 gcallis <@t> montana.edu Tue May 4 13:33:01 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] CJD sureveillance center correct web address In-Reply-To: <5.1.0.14.0.20040428105750.00b858f0@pop.cwru.edu> Message-ID: <3.0.6.32.20040504123301.00bf6a80@gemini.msu.montana.edu> Sorry, but - The correct website Address http://www.cjdsurveillance.com --not org. At 11:03 AM 4/28/2004 -0400, you wrote: >The National Prion Disease Pathology Surveillance Center >(www.cjdsurvelliance.org) provides a full service to handle suspected CJD >cases. You can call 216--368-0587 or visit our website > >Phyllis Scalzo, HT(ASCP) >Case Western Reserve University >2085 Adelbert Rd. >Cleveland, Ohio 44106 >Ph: 216-368-0822 >Fax: 216-368-2546 > > >_______________________________________________ >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 epitek <@t> yahoo.com Tue May 4 14:57:01 2004 From: epitek <@t> yahoo.com (Elaine Pitek) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] unsubscribe In-Reply-To: Message-ID: <20040504195701.82234.qmail@web41606.mail.yahoo.com> marilyn.j.easton@gsk.com wrote: Sounds like a problem with your water bath temperature. Too many wrinkles means your water bath is too cold and quick separation is an indication that your water bath is too hot or you have a processing problem. We generally use a 38 to 42 degree centigrade water bath temperature depending on how quickly you can pick up your section. The rat cerebellum tends to separate more quickly than the cerebrum which is why we block the tissue so that the cerebellum is picked up first on the slide and the cerebrum can be "dangled" in the waterbath a few seconds longer if needed. Processing plays a big part in how well your brain tissue sections, so you should try picking up your sections at different temperatures to get a feel for what works best with your tissue. Marilyn Easton, B.A., HT/HTL (ASCP), LAT (AALAS) Scientist, Pathology Dept., Safety Assessment GlaxoSmithKline 919-483-7891 "David Clark" Sent by: To: Histonet@lists.utsouthwestern.edu histonet-bounces@lists.utsouth western.edu cc: Subject: [Histonet] cutting rat brains 03-May-2004 09:39 Hello, I was wandering if any of you wonderful Histonet people who have much experience cutting rat brains can offer some assistance in trying to cut good quality sections. We either encounter lots of wrinkles or tissue seperation from the paraffin sections once placed on the waterbath. Is this a processing problem or do you think it is strictly a sectioning problem? Any suggestions would be appreciated. Thanks, David Clark Neuropathology OHSU _______________________________________________ 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 Elaine Pitek APMG/Ameripath 408-399-5050 x 123 --------------------------------- Do you Yahoo!? Win a $20,000 Career Makeover at Yahoo! HotJobs From jkiernan <@t> uwo.ca Tue May 4 16:27:42 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Luxol Fast blue References: <1083688413.4097c5dd102f9@webmail.umdnj.edu> Message-ID: <40980ACE.C88B0E9A@uwo.ca> Luxol fast blue works well on frozen sections. You may need a longer differentiation in 0.05% lithium carbonate than with paraffin sections. Neutral red provides a contrasting Nissl stain and also darkens the myelin already coloured by the luxol fast blue. John Kiernan London, Canada _____________________________________ Michael Kurnellas wrote: > > I was told that Luxol fast blue staining should be done on paraffin embedded > tissue. Can I also do this stain on sucrose-embedded tissue, frozen, and > then placed in OCT compound before 10um sections were taken? > > Thanks. > Mike K. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SJones <@t> cvm.tamu.edu Tue May 4 19:00:13 2004 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Mouse and rat injured spinal cord - polyester wax Message-ID: I tried polyester wax years ago and came to the conclusion I needed to be cutting in a walk-in-cooler. I never thought of keeping the knife and block in the fridge, but it make sense. I second the vote for any other method. Unless you love challenges and have the patience of a saint. Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> 5/4/2004 10:44:42 AM >>> Hi Rebecca, Many, many years ago I used 37 degree C MPT polyester wax. There are several major problems using the stuff. Key one is temperature. Blocks and knife were kept in the fridge, with both brought out just prior to use -we used a Cambridge rocker which meant transfer of knife to and from the fridge was easy. In the summer in our lab which had no air conditioning, there are minutes of cutting time available before the block and kife warms up and sectioning has to stop. One paper I read partly overcame this narrow curring window by using a flower sieve packed with dry ice suspended above the microtome. our resources did not stretch that far in those days (mid 70's). Once you have your sections, we had to use a starch based adhesive which was added to the slide and sections floated directly onto the pool of adesive. The trick was to ensure one end of the ribbon overlapped the end of the slide so that after the short period of time the sections took to expand and de-crease, the slide could be tipped allowing the adhesive to run away leaving a nice ribbon on the slide. The rest was easy!! Having read all of this, it comes as no surprise that my advice is to try every other option for getting what you want first. Best of luck. Steve Rebecca Nishi Sent by: To: histonet-bounces@lists.utsouth cc: western.edu Subject: [Histonet] Mouse and rat injured spinal cord - polyester wax 04/05/2004 16:19 Has anyone used polyester wax embedding for mouse or rat injured spinal cord. We would like to use a low melt wax instead of paraffin or plastic to preserve some of the antigens we are interested in, but we also are interested in doing stereology, so we want consistent and a complete set of sections. Also, the injured area poses many problems for sectioning. Does anyone have any tips for cutting and staining with this wax? I am going to use a rotary microtome. Thanks Rebecca _______________________________________________ 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 balajimr <@t> drreddys.com Tue May 4 22:26:07 2004 From: balajimr <@t> drreddys.com (balajimr@drreddys.com) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Basics - IHC Message-ID: I have to prepare for an inhouse seminar on basics of immunohistochemistry and its aplications in research. Could anyone suggest me best/ideal literature/reference available on the web for the preparationof the same. Balaji From arme <@t> optonline.net Wed May 5 00:20:52 2004 From: arme <@t> optonline.net (Mark Sofferman) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Tissue processor available In-Reply-To: <0HX30032QJ7IAA@mta14.srv.hcvlny.cv.net> Message-ID: We have available a Fisher MP166 Processor. Email if interested. Mark American Resource Tel. 201-833-1550 ----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Sunday, May 02, 2004 1:00 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 6, Issue 2 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. Re: nail sectioning (jhnspam@aol.com) ---------------------------------------------------------------------- Message: 1 Date: Sat, 1 May 2004 23:23:25 EDT From: jhnspam@aol.com Subject: Re: [Histonet] nail sectioning To: mary.bliss@northwestpathology.com, histonet@pathology.swmed.edu Message-ID: <85.acd226e.2dc5c3ad@aol.com> Content-Type: text/plain; charset="US-ASCII" We face in to the block and then soak it in Downy Fabric Softener for about 5 minutes. Put the section on a plus slide and it comes out beautiful. We do PAS/LG on all nails and they look great as well. Pam Johnson ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 6, Issue 2 ************************************** From jkiernan <@t> uwo.ca Wed May 5 00:23:34 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Polyester wax (with earlier citations) References: Message-ID: <40987A56.FD17E367@uwo.ca> 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? -- ------------------------------- 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 _______________________________ Sarah Jones wrote: > > I tried polyester wax years ago and came to the conclusion I needed to > be cutting in a walk-in-cooler. I never thought of keeping the knife > and block in the fridge, but it make sense. I second the vote for any > other method. Unless you love challenges and have the patience of a > saint. > > Sarah Jones HT(ASCP) > Dept. of Vet. Anatomy & Public Health > Histology Lab > Texas A&M University > College Station, TX 77843-4458 > phone: 979-845-3177 > fax: 979-458-3499 > > >>> 5/4/2004 10:44:42 AM >>> > > Hi Rebecca, > > Many, many years ago I used 37 degree C MPT polyester wax. There are > several major problems using the stuff. Key one is temperature. Blocks > and > knife were kept in the fridge, with both brought out just prior to use > -we > used a Cambridge rocker which meant transfer of knife to and from the > fridge was easy. In the summer in our lab which had no air > conditioning, > there are minutes of cutting time available before the block and kife > warms > up and sectioning has to stop. One paper I read partly overcame this > narrow > curring window by using a flower sieve packed with dry ice suspended > above > the microtome. our resources did not stretch that far in those days > (mid > 70's). Once you have your sections, we had to use a starch based > adhesive > which was added to the slide and sections floated directly onto the > pool of > adesive. The trick was to ensure one end of the ribbon overlapped the > end > of the slide so that after the short period of time the sections took > to > expand and de-crease, the slide could be tipped allowing the adhesive > to > run away leaving a nice ribbon on the slide. The rest was easy!! > > Having read all of this, it comes as no surprise that my advice is to > try > every other option for getting what you want first. > > Best of luck. > > Steve > > > > > Rebecca Nishi > > > Sent by: To: > > > histonet-bounces@lists.utsouth cc: > > > western.edu Subject: > [Histonet] Mouse and rat injured spinal cord - polyester wax > > > > > > > > 04/05/2004 16:19 > > > > > > > > > > Has anyone used polyester wax embedding for mouse or rat injured > spinal > cord. We would like to use a low melt wax instead of paraffin or > plastic to > preserve some of the antigens we are interested in, but we also are > interested in doing stereology, so we want consistent and a complete > set of > sections. Also, the injured area poses many problems for sectioning. > > Does anyone have any tips for cutting and staining with this wax? I am > going > to use a rotary microtome. > > Thanks Rebecca > > _______________________________________________ > 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 Stephen.Eyres <@t> sanofi-synthelabo.com Wed May 5 04:11:40 2004 From: Stephen.Eyres <@t> sanofi-synthelabo.com (Stephen.Eyres@sanofi-synthelabo.com) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Re: Polyester wax (with earlier citations) Message-ID: Hi John, I don't think there are any reasons for using it now. When I used it in the 70's it was because one of our pathologists wanted to cut thin sections from biopsies. The sections were good when you eventually produced any!! I dabbled with Digol Distrearate for a while but found sectioning this difficult too. Then along came methacrylate and the rest, as they say, is history. The Pathologist in question was a John Southgate who had a lab in his office for dabbling in histology procedures. He obviously got the technical bug from his microbiologist father, of Southgate's Mucicarmine fame. John Southgate also developed a Zenker-Susa fixative procedure for biosy fixing which involved giving two pots, one for Zenker, and one for Susa, to the nurse, who mixed both immediately prior to adding the biopsy. There was then a max of 4 hrs fixation (if I recall correctly), followed by a water wash to remove mercury. Processing was on a Technicon Ultra, with it's oil heated bath. Embedding - this was before we had plastic cassettes -was done using ice cube trays filled with hot wax from a small teapot sitting on a stand heated with a bunsen burner. Specimen identification was on small paper strip, the end of which was inserted into the cooling wax block. A full tray was floated onto cold water and when the surface wax had solidified, the whole tray was immersed. The times the labels would separate from the wax block and time was often wasted matching up unlabelled blocks with paper labels. Then, a small guilotine was used to coarsly trim away wax excess wax, before a flat blades spatular-type heated iron was used to finely create the final block. The block was attached to a wooden block which had it's surface heated by the heated iron. The specimen label was attached to the wooden block and the heated iron was run over the back of the wax block, with the hot wax allowed to drip onto the wooden block and the wax block applied very quickly. It is not hard to imagine the frustration of experienced microtomists when trimming in blocked caused many to fall off indicating a trainee was learning this process and blocks were not attached properly. And then the blocks had to be removed later using a 'blunt' knife. Ahhhh, they were the days, when many a day I'd go home with brown hands after making up 10% formal saline, the smell of which was eased by the gentle aroma of pipe tobacco wafting around the lab from the pathologist who was performing a cut-up (grossing). I cannot remember hearing the word safety then. Well better get back to work. Cheers Steve John Kiernan > cc: histonet-bounces@lists.utsouthwestern.edu Stephen Eyres/GB-ALNWICK/RESEARCH/SANOFI@Research 05/05/2004 06:23 rnishi@uci.edu Histonet@lists.utsouthwestern.edu Subject: Polyester wax (with earlier citations) 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? -- ------------------------------- 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 _______________________________ Sarah Jones wrote: > > I tried polyester wax years ago and came to the conclusion I needed to > be cutting in a walk-in-cooler. I never thought of keeping the knife > and block in the fridge, but it make sense. I second the vote for any > other method. Unless you love challenges and have the patience of a > saint. > > Sarah Jones HT(ASCP) > Dept. of Vet. Anatomy & Public Health > Histology Lab > Texas A&M University > College Station, TX 77843-4458 > phone: 979-845-3177 > fax: 979-458-3499 > > >>> 5/4/2004 10:44:42 AM >>> > > Hi Rebecca, > > Many, many years ago I used 37 degree C MPT polyester wax. There are > several major problems using the stuff. Key one is temperature. Blocks > and > knife were kept in the fridge, with both brought out just prior to use > -we > used a Cambridge rocker which meant transfer of knife to and from the > fridge was easy. In the summer in our lab which had no air > conditioning, > there are minutes of cutting time available before the block and kife > warms > up and sectioning has to stop. One paper I read partly overcame this > narrow > curring window by using a flower sieve packed with dry ice suspended > above > the microtome. our resources did not stretch that far in those days > (mid > 70's). Once you have your sections, we had to use a starch based > adhesive > which was added to the slide and sections floated directly onto the > pool of > adesive. The trick was to ensure one end of the ribbon overlapped the > end > of the slide so that after the short period of time the sections took > to > expand and de-crease, the slide could be tipped allowing the adhesive > to > run away leaving a nice ribbon on the slide. The rest was easy!! > > Having read all of this, it comes as no surprise that my advice is to > try > every other option for getting what you want first. > > Best of luck. > > Steve > > > > > Rebecca Nishi > > > Sent by: To: > > > histonet-bounces@lists.utsouth cc: > > > western.edu Subject: > [Histonet] Mouse and rat injured spinal cord - polyester wax > > > > > > > > 04/05/2004 16:19 > > > > > > > > > > Has anyone used polyester wax embedding for mouse or rat injured > spinal > cord. We would like to use a low melt wax instead of paraffin or > plastic to > preserve some of the antigens we are interested in, but we also are > interested in doing stereology, so we want consistent and a complete > set of > sections. Also, the injured area poses many problems for sectioning. > > Does anyone have any tips for cutting and staining with this wax? I am > going > to use a rotary microtome. > > Thanks Rebecca > > _______________________________________________ > 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 pstefanova <@t> sten.sunnybrook.utoronto.ca Wed May 5 08:08:20 2004 From: pstefanova <@t> sten.sunnybrook.utoronto.ca (Petia P Stefanova) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Basics - IHC References: Message-ID: <003501c432a2$0edbc0b0$9d194c8e@WS21203> Hi Balaji, you can find information about basic IHC on these links. http://www.chemicon.com/resource/ANT101/a2D.asp http://sciencepark.mdanderson.org/histology/ihc.html http://www.protocol-online.org/prot/Immunology/Immunohistochemistry/index.html Good luck! Petia From JMcCormi <@t> schosp.org Wed May 5 08:59:36 2004 From: JMcCormi <@t> schosp.org (McCormick, James) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Re: Polyester wax (with earlier citations),and yet EARLIER references Message-ID: <229A3566B9F0D311826E00D0B7441D7905E2FCC7@swedish_nt1.schosp.org> John,Stephen,Sarah,Rebecca et al, Your notes and comments on the histoNet re embedding media caught my eye!! As a practicing pathologist of some years experience, and a histotechnonogist at heart, I have worked with the opportunities (problems) of tissue embedding and sectioning since my first student years at the university of Notre Dame where we used as a text book "Animal Micrology" Geyer 4th edition 1936,. This book first introduced me to the systems of preparing tissues for embedding and sectioning. Truly, not much has changed. In fact, a further reference is "the microtomist's Vade Mecum" by Arthur Bolles Lee, Churchill, London 1885 page 170 #223. I can't say "I've done it all" , but almost : paraffin, celloidin,latex double embedding,Steedman's polyester,epoxy,methacrolate,Frozen sections,etc..,etc.....Bracegirdle "a history of Microtechnique" relates most of the history and evolution of materials and methods for slide making in the natural sciences....all, as developed from need. I continue to search for "a better everything for Histotechnology" and enjoy walking with you to identify new methods and materials. For the moment: A well fixed tissue, completely dehydrated,cleared and infiltrated with paraffin (as Lee said in 1885,"the paraffin slab/grade should ring when you thump it"), a sharp knife at the correct cutting angle and a well trained technician to turn the crank,(or slide) WILL MOST CERTAINLY PRODUCE THE DESIRED RESULT. Because of your collective interest and wisdom I would most enthusiastically recommend the historical reference books. (As a service and hobby,Mrs. McCormick and I have published these books as a reference resource from the Science Heritage Collection) WWW.scienceheritage.com . Kindest regards, J.B.McCormick M.D., Pathologist,Swedish Covenant Hospital , Chicago -----Original Message----- From: Stephen.Eyres@sanofi-synthelabo.com [mailto:Stephen.Eyres@sanofi-synthelabo.com] Sent: Wednesday, May 05, 2004 4:12 AM To: John Kiernan Cc: Histonet@lists.utsouthwestern.edu; rnishi@uci.edu; Sarah Jones; histonet-bounces@lists.utsouthwestern.edu Subject: [Histonet] Re: Polyester wax (with earlier citations) Hi John, I don't think there are any reasons for using it now. When I used it in the 70's it was because one of our pathologists wanted to cut thin sections from biopsies. The sections were good when you eventually produced any!! I dabbled with Digol Distrearate for a while but found sectioning this difficult too. Then along came methacrylate and the rest, as they say, is history. The Pathologist in question was a John Southgate who had a lab in his office for dabbling in histology procedures. He obviously got the technical bug from his microbiologist father, of Southgate's Mucicarmine fame. John Southgate also developed a Zenker-Susa fixative procedure for biosy fixing which involved giving two pots, one for Zenker, and one for Susa, to the nurse, who mixed both immediately prior to adding the biopsy. There was then a max of 4 hrs fixation (if I recall correctly), followed by a water wash to remove mercury. Processing was on a Technicon Ultra, with it's oil heated bath. Embedding - this was before we had plastic cassettes -was done using ice cube trays filled with hot wax from a small teapot sitting on a stand heated with a bunsen burner. Specimen identification was on small paper strip, the end of which was inserted into the cooling wax block. A full tray was floated onto cold water and when the surface wax had solidified, the whole tray was immersed. The times the labels would separate from the wax block and time was often wasted matching up unlabelled blocks with paper labels. Then, a small guilotine was used to coarsly trim away wax excess wax, before a flat blades spatular-type heated iron was used to finely create the final block. The block was attached to a wooden block which had it's surface heated by the heated iron. The specimen label was attached to the wooden block and the heated iron was run over the back of the wax block, with the hot wax allowed to drip onto the wooden block and the wax block applied very quickly. It is not hard to imagine the frustration of experienced microtomists when trimming in blocked caused many to fall off indicating a trainee was learning this process and blocks were not attached properly. And then the blocks had to be removed later using a 'blunt' knife. Ahhhh, they were the days, when many a day I'd go home with brown hands after making up 10% formal saline, the smell of which was eased by the gentle aroma of pipe tobacco wafting around the lab from the pathologist who was performing a cut-up (grossing). I cannot remember hearing the word safety then. Well better get back to work. Cheers Steve John Kiernan > cc: histonet-bounces@lists.utsouthwestern.edu Stephen Eyres/GB-ALNWICK/RESEARCH/SANOFI@Research 05/05/2004 06:23 rnishi@uci.edu Histonet@lists.utsouthwestern.edu Subject: Polyester wax (with earlier citations) 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? -- ------------------------------- 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 _______________________________ Sarah Jones wrote: > > I tried polyester wax years ago and came to the conclusion I needed to > be cutting in a walk-in-cooler. I never thought of keeping the knife > and block in the fridge, but it make sense. I second the vote for any > other method. Unless you love challenges and have the patience of a > saint. > > Sarah Jones HT(ASCP) > Dept. of Vet. Anatomy & Public Health > Histology Lab > Texas A&M University > College Station, TX 77843-4458 > phone: 979-845-3177 > fax: 979-458-3499 > > >>> 5/4/2004 10:44:42 AM >>> > > Hi Rebecca, > > Many, many years ago I used 37 degree C MPT polyester wax. There are > several major problems using the stuff. Key one is temperature. Blocks > and > knife were kept in the fridge, with both brought out just prior to use > -we > used a Cambridge rocker which meant transfer of knife to and from the > fridge was easy. In the summer in our lab which had no air > conditioning, > there are minutes of cutting time available before the block and kife > warms > up and sectioning has to stop. One paper I read partly overcame this > narrow > curring window by using a flower sieve packed with dry ice suspended > above > the microtome. our resources did not stretch that far in those days > (mid > 70's). Once you have your sections, we had to use a starch based > adhesive > which was added to the slide and sections floated directly onto the > pool of > adesive. The trick was to ensure one end of the ribbon overlapped the > end > of the slide so that after the short period of time the sections took > to > expand and de-crease, the slide could be tipped allowing the adhesive > to > run away leaving a nice ribbon on the slide. The rest was easy!! > > Having read all of this, it comes as no surprise that my advice is to > try > every other option for getting what you want first. > > Best of luck. > > Steve > > > > > Rebecca Nishi > > > Sent by: To: > > > histonet-bounces@lists.utsouth cc: > > > western.edu Subject: > [Histonet] Mouse and rat injured spinal cord - polyester wax > > > > > > > > 04/05/2004 16:19 > > > > > > > > > > Has anyone used polyester wax embedding for mouse or rat injured > spinal > cord. We would like to use a low melt wax instead of paraffin or > plastic to > preserve some of the antigens we are interested in, but we also are > interested in doing stereology, so we want consistent and a complete > set of > sections. Also, the injured area poses many problems for sectioning. > > Does anyone have any tips for cutting and staining with this wax? I am > going > to use a rotary microtome. > > Thanks Rebecca > > _______________________________________________ > 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 *** 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 sharon.willman <@t> bms.com Wed May 5 09:08:56 2004 From: sharon.willman <@t> bms.com (Sharon E Willman) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] S-100 Message-ID: <4098F578.E41A95ED@bms.com> Hi, I appreciate the replies on my question regarding S-100. I do have another question for those who have worked with S-100. When doing S-100, do you have collagen and adipose tissue that stains dark? The positive and negative controls turn out nicely. Thank you for any additional help! Sharon Willman From vandries <@t> vub.ac.be Wed May 5 09:19:08 2004 From: vandries <@t> vub.ac.be (Veronique Andriessen) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] double immuno staining Message-ID: Hi folks, I need some help. I am struggling with a double immunostaining for somatostatin receptor and desmin. All my previous double stainings worked beautifully, but they were performed on fresh frozen acetone fixed pancreatic tissue. For staining of somatostatin receptor I have to fix with formaldehyde. I use 20 minutes 0.75% PFA perfused tissue which is snap frozen embedded in tissue freezing medium and cut on a cryostat. I was advised to do it this way to prevent loss of antigenicity. Morphology is quite good. This fixation works for the somatostatin receptor staining, but it kills the desmin signal. I was able to retrieve most of the desmin signal by EIER for 10 minutes with trypsin-EDTA, but now the morphology is awful. Can someone give me some advice? Sincerely yours, Veronique Andriessen BAS Lab. Molecular Liver Cell Biology, Free University Brussels (VUB)Belgium From dobbin <@t> upei.ca Wed May 5 09:30:59 2004 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] double immuno staining In-Reply-To: Message-ID: <4098D073.26025.924778@localhost> Hi Veronique, I would suggest you are already on the right track! Perhaps just shorten the EIER time, try a different enzyme or try a heat-induced epitope retreival method. Good luck. Greg From: "Veronique Andriessen" To: "Histonet" Date sent: Wed, 5 May 2004 16:19:08 +0200 Subject: [Histonet] double immuno staining > Hi folks, > > I need some help. > > I am struggling with a double immunostaining for somatostatin receptor and > desmin. > > All my previous double stainings worked beautifully, but they were performed > on fresh frozen acetone fixed pancreatic tissue. > > For staining of somatostatin receptor I have to fix with formaldehyde. I use > 20 minutes 0.75% PFA perfused tissue which is snap frozen embedded in > tissue freezing medium and cut on a cryostat. I was advised to do it this > way to prevent loss of antigenicity. > Morphology is quite good. > This fixation works for the somatostatin receptor staining, but it kills the > desmin signal. > I was able to retrieve most of the desmin signal by EIER for 10 minutes with > trypsin-EDTA, but now the morphology is awful. > > Can someone give me some advice? > > Sincerely yours, > > Veronique Andriessen BAS > Lab. Molecular Liver Cell Biology, > Free University Brussels (VUB)Belgium > > > > _______________________________________________ > 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 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Making a living is getting; making a life is giving. From SCheasty <@t> ahs.llumc.edu Wed May 5 09:57:32 2004 From: SCheasty <@t> ahs.llumc.edu (Cheasty, Sandra) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Lab Storage - floor pallettes Message-ID: <2E50F33F91EEDA46A77BC3B2575BB09121E0C1@mars.llumc.edu> Does anyone out there have a source for floor palettes that are only 24" deep? The only ones I can find so far are all 40" deep. (We need to store some supplies off the floor under the counters.) Thank you, Sandy Confidentiality Note: The preceding e-mail message (including any attachments) contains information that may be confidential, protected by applicable legal privileges, or constitute non-public information. It is intended to be conveyed only to the designated recipient(s). If you are not an intended recipient of this message, please notify the sender by replying to this message and then delete it from your system. Use, dissemination, distribution or reproduction of this message by unintended recipients is not authorized and may be unlawful. From pstefanova <@t> sten.sunnybrook.utoronto.ca Wed May 5 10:02:57 2004 From: pstefanova <@t> sten.sunnybrook.utoronto.ca (Petia P Stefanova) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] cutting rat pancreas References: Message-ID: <005a01c432b2$0d722b50$9d194c8e@WS21203> Hi, I am trying to cut good quality sections from rat pancreas but I get lots of wrinkles. My other tissues are great just the pancreas makes problems. I guess the problem could be from the processing program. I would very much appreciate your help. Thanks to all! Petia From gsennello <@t> osip.com Wed May 5 10:10:29 2004 From: gsennello <@t> osip.com (Sennello, Gina) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Lab Storage - floor pallets Message-ID: A company called Seton has them 25 inches deep. Their number is 1-800-243-6624 Gina Sennello Senior Associate Scientist Histotechnologist OSIP 2860 Wilderness Place Boulder, CO 80301 phone 303-546-7739 fax 303-444-0672 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Cheasty, Sandra Sent: Wednesday, May 05, 2004 8:58 AM To: HistoNet (E-mail) Subject: [Histonet] Lab Storage - floor pallettes Does anyone out there have a source for floor palettes that are only 24" deep? The only ones I can find so far are all 40" deep. (We need to store some supplies off the floor under the counters.) Thank you, Sandy Confidentiality Note: The preceding e-mail message (including any attachments) contains information that may be confidential, protected by applicable legal privileges, or constitute non-public information. It is intended to be conveyed only to the designated recipient(s). If you are not an intended recipient of this message, please notify the sender by replying to this message and then delete it from your system. Use, dissemination, distribution or reproduction of this message by unintended recipients is not authorized and may be unlawful. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Stephen.Spencer <@t> intervet.com Wed May 5 10:21:42 2004 From: Stephen.Spencer <@t> intervet.com (Spencer, S (Stephen)) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Vacancy - Veterinary Histopathology UK Message-ID: <6BE0EFB134CAD611B0A400408102ECD14E1D56@mksn71.mks.intra> Colleagues, May I bring to your attention the following vacancy which has been created by the relocation of our Houghton Lab to Milton Keynes. Please contact me by phone or email for further details. Stephen Spencer Research Veterinarian Intervet UK 01480 830009 ____________________________________________________________________________ ________ Scientist - Veterinary Histopathology Milton Keynes Intervet International BV, part of the Akzo Nobel Group, is a highly successful animal health company, currently the world's third largest. We have nearly 5000 employees throughout the world, of which 750 are engaged in research and development. Here in the UK, we are in the process of creating one of the country's largest bacteriology and virology research centres, which within the next 12 months will employ up to 80 scientists and support staff. The successful applicant will be an important member of the Biological Services Department, providing histopathology support for our veterinary research programme. Primary responsibilities will include the organisation and running of the histopathology laboratory, including tissue processing, microtomy, cryotomy, routine and special staining, immunohistochemistry and light microscopy. This will involve planning and scheduling laboratory activities in consultation with Project Managers and in response to project demands. We are looking for a scientist who has histopathological experience and possesses a good working knowledge of relevant laboratory techniques. Self-motivation and good organisational skills are essential. Previous experience of working in an environment compliant with the principles of GLP would be advantageous. Knowledge or an interest in pathology associated with avian and animal diseases would be desirable, but is not essential. This position is based at our newly refurbished facility in Milton Keynes. We offer a highly attractive benefits package, which includes a competitive salary, private healthcare cover, contributory pension scheme and life assurance as well as on-site sports and social facilities. CPD participation is actively encouraged. For further information and an informal chat about this role, please telephone Stephen Spencer, on 01908 665050. If you wish to apply for this position, please telephone our Human Resources department on 01908 685216 for an application pack. Closing Date: 19th May. Only applicants with the right to live and work in the UK will be considered for this position and proof may be requested. _________________________ Stephen Spencer Research Veterinarian BSD - Intervet UK The Farm Tel; 01480 830009 -------------------------------------- This message, including attachments, is confidential and may be privileged. If you are not an intended recipient, please notify the sender then delete and destroy the original message and all copies. You should not copy, forward and/or disclose this message, in whole or in part, without permission of the sender. -------------------------------------- From M.Bromley <@t> dgri.scot.nhs.uk Wed May 5 10:25:46 2004 From: M.Bromley <@t> dgri.scot.nhs.uk (Mike Bromley) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] lymph nodes Message-ID: <7325D637DFE2D211928800902733A7F303B94ACC@DGAMTBDCEMS> Hi Everyone I am after a very thin blade to slice unfixed lymph nodes, does anyone have any ideas? Ideally some kind of holder for old style razor blades would be the answer. Best Wishes Mike Bromley Chief Biomedical Scientist Pathology Dumfries & Galloway Royal Infirmary Scotland, UK From Terry.Marshall <@t> rothgen.nhs.uk Wed May 5 10:36:42 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] lymph nodes Message-ID: I find for this sort of thing, disposable microtome blades are ideal. Holders are available, but I neither use nor recommend them. 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: Mike Bromley [mailto:M.Bromley@dgri.scot.nhs.uk] Sent: 05 May 2004 16:26 To: Histonet (E-mail) Subject: [Histonet] lymph nodes Hi Everyone I am after a very thin blade to slice unfixed lymph nodes, does anyone have any ideas? Ideally some kind of holder for old style razor blades would be the answer. Best Wishes Mike Bromley Chief Biomedical Scientist Pathology Dumfries & Galloway Royal Infirmary Scotland, UK _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rnishi <@t> uci.edu Wed May 5 10:54:54 2004 From: rnishi <@t> uci.edu (Rebecca Nishi) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Re: Polyester wax (with earlier citations)-it does work In-Reply-To: <40987A56.FD17E367@uwo.ca> Message-ID: 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? From gcallis <@t> montana.edu Wed May 5 10:54:56 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] double immuno staining In-Reply-To: Message-ID: <3.0.6.32.20040505095456.00c015d8@gemini.msu.montana.edu> It probably is your fixation which is compatible with one antibody but not the other. Formalin is the culprit and you still "lose" antigenicity due to cross linking of antigen by the formaldehyde and paraformaldehyde. You may want to try perfusion with PLP (something one lab does in our department) then post fix in that fixative. Snap freezing of fresh tissue DOES preserve antigens, so you can try unfixed tissue, snap frozen, then fixed with other fixative AFTER you cut overnight RT air dried frozen sections. If you use acetone, 4C for 10 min or another fixative choice, a fixation panel, antigen retrieval is not needed, and you may have far more success with both antibodies. Antigen retrieval and enzyme digestion (also using strong peroxidase blocking - chews sections off slides at times) methods on frozen sections can damage morphology, so you may want to work with a fixative where you never have to do any retrieval whatsoever. Chris van der Loos may respond to this message also and he is the master of multiple staining. At 04:19 PM 5/5/2004 +0200, you wrote: >Hi folks, > >I need some help. > >I am struggling with a double immunostaining for somatostatin receptor and >desmin. > >All my previous double stainings worked beautifully, but they were performed >on fresh frozen acetone fixed pancreatic tissue. > >For staining of somatostatin receptor I have to fix with formaldehyde. I use >20 minutes 0.75% PFA perfused tissue which is snap frozen embedded in >tissue freezing medium and cut on a cryostat. I was advised to do it this >way to prevent loss of antigenicity. >Morphology is quite good. >This fixation works for the somatostatin receptor staining, but it kills the >desmin signal. >I was able to retrieve most of the desmin signal by EIER for 10 minutes with >trypsin-EDTA, but now the morphology is awful. > >Can someone give me some advice? > >Sincerely yours, > >Veronique Andriessen BAS >Lab. Molecular Liver Cell Biology, >Free University Brussels (VUB)Belgium > > > >_______________________________________________ >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 John.Auld <@t> whnt.nhs.uk Wed May 5 11:39:01 2004 From: John.Auld <@t> whnt.nhs.uk (John Auld) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Triple G and Slide Writers Message-ID: Hi All Is anyone using Triple G computer system with a slide writer? We are thinking of going down this route and wonder if anyone has any experiences they would share with us. Many thanks in advance John John Auld Dept of Histopathology and Clinical Cytology Arrowe Park Hospital Arrowe Park Road Upton Wirral From convmcm <@t> cc.usu.edu Wed May 5 12:09:25 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Basics - IHC In-Reply-To: Message-ID: <000001c432c3$b91f3c20$4a737b81@Cygnus> DAKO Cytomation has the best handbook in PDF format on IHC and it's free on the web. Got to www.dakocytomation.com. Also US Biological has a PDF publication called Introduction to antibodies that seems pretty good, too. I just downloaded it today, so I can't say what it's like, but I suppose it will be a good reference. Cheers. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of balajimr@drreddys.com Sent: Tuesday, May 04, 2004 8:26 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Basics - IHC Importance: High I have to prepare for an inhouse seminar on basics of immunohistochemistry and its aplications in research. Could anyone suggest me best/ideal literature/reference available on the web for the preparationof the same. Balaji _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DonnaWillis <@t> texashealth.org Wed May 5 12:24:53 2004 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Cerner Millennium Message-ID: <5C6CBCCEB04B894C8BD15B312487F7B201A330C4@ftwex01.txhealth.org> We are building the Cerner Millenium system and looking to talk with other user of this system about their experiences with slide and cassette labelers. We are trying to get the Leica labelers interfaced with Millenium. Please email me with your experiences. Thanks, Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx 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 kgrobert <@t> rci.rutgers.edu Wed May 5 12:54:18 2004 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Grocott's Methenamine Silver Stain Message-ID: <40992A4A.3000604@rci.rutgers.edu> Hey, all- I've been trying to stain some FFPE slides of decalcified rat sinuses for fungi using the AFIP protocol for Grocott's stain, and despite using fresh reagents, the positive control and unknowns do not stain at all, except for some random deposition of silver that does not even begin to look like fungi. The positive control is loaded with Candida, and can be seen with H&E easily. So far, we don't see any fungi in the rat sinuses and lungs (they were sneezing, and the bedding came back positive for Aspergillus) with H&E staining, which is why we decided to try the Grocott's-one way or another we should be able to visualize it in the tissues. Any ideas why the Candida-loaded positive control would not stain with Grocott's? Right now I am thinking that perhaps they didn't sit long enough in the 4% chromic acid, even though the protocol says to soak the slides for an hour to oxidize them. Either that or a longer soak in the methenamine-silver solution at 60 degrees C (also for one hour). Thanks in advance for your help- Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Dept of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers University 41 B Gordon Rd Piscataway, NJ 08854 From LGORMAN1 <@t> kumc.edu Wed May 5 12:50:10 2004 From: LGORMAN1 <@t> kumc.edu (LINDA GORMAN) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Streck Fixative Message-ID: Can someone give me the information to order Streck fixative? Thanks,Linda From ryaskovich <@t> dir.nidcr.nih.gov Wed May 5 13:20:14 2004 From: ryaskovich <@t> dir.nidcr.nih.gov (Yaskovich, Ruth A (NIH/NIDCR)) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Streck Fixative Message-ID: <8F3AB322628548428A992EFB0E80F5D3A2F027@nihexchange8.nih.gov> Linda, The name is Streck Laboratories 7002 S 109 St. La Vistal NE. 68128. There is a web site Ruth Yaskovich N.I.H. > ---------- > From: LINDA GORMAN > Sent: Wednesday, May 5, 2004 12:50 PM > To: histonet@pathology.swmed.edu > Subject: [Histonet] Streck Fixative > > Can someone give me the information to order Streck fixative? > Thanks,Linda > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From RichmanL <@t> MedImmune.com Wed May 5 13:26:56 2004 From: RichmanL <@t> MedImmune.com (Richman, Laura) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] histotech opportunity Message-ID: <83899F0EC7671543B305FB5694024DE603482A1B@medimmune4.medimmune.com> MedImmune, Inc., a publicly owned biotech company, is seeking an HT or HTL (ASCP) certified candidate who possess excellent histology skills to assist in running a histopathology lab. Successful candidates will be responsible for most aspects of routine histological procedures including paraffin processing, sectioning of both paraffin and frozen blocks, basic stains such as H&E and Giemsa, limited special stains, and training of other lab personnel. GLP and supervisory experience helpful. Individual must be highly motivated and independent, with strong organizational skills. MedImmune, Inc. has comprehensive benefits package and excellent opportunities for advancement and career growth. The position location is at our new headquarters in Gaithersburg, Maryland. Please send your CV or direct questions to: Laura K. Richman, DVM, PhD Diplomate, ACVP Pathologist/Scientist II MedImmune, Inc. One MedImmune Way Gaithersburg, MD 20878 direct dial: (301) 398-4741 e-fax (301) 398-9741 RichmanL@MedImmune.com From kgrobert <@t> rci.rutgers.edu Wed May 5 14:15:50 2004 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Grocott's Methenamine Silver Stain In-Reply-To: References: Message-ID: <40993D66.8070503@rci.rutgers.edu> I am not sure how long they were in decal...but I supplied some of our Fisher Scientific Cal-EX, which has hydrochloric acid and EDTA in it, to the Rutgers vet who had this case of sneezing rats. I did ask him how long he decal'd them, but he doesn't remember either. The chromic acid was freshly ordered as a 10% solution from LabChem, Inc., which was then diluted down to 4% the morning I was to use it. Thanks for your help! Kathleen Bartlett, Jeanine wrote: > Kathleen: > > How long were they in decal and what type of decal? And do you make > your chromic fresh? > > Jeanine Bartlett, HT(ASCP) > Centers for Disease Control > Infectious Disease Pathology Activity > 1600 Clifton Road, MS/G-32 > Atlanta, GA 30333 > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kathleen > Roberts > Sent: Wednesday, May 05, 2004 1:54 PM > To: 'histonet@lists.utsouthwestern.edu' > Subject: [Histonet] Grocott's Methenamine Silver Stain > > > Hey, all- > > I've been trying to stain some FFPE slides of decalcified rat sinuses > for fungi using the AFIP protocol for Grocott's stain, and despite using > > fresh reagents, the positive control and unknowns do not stain at all, > except for some random deposition of silver that does not even begin to > look like fungi. The positive control is loaded with Candida, and can > be seen with H&E easily. > > So far, we don't see any fungi in the rat sinuses and lungs (they were > sneezing, and the bedding came back positive for Aspergillus) with H&E > staining, which is why we decided to try the Grocott's-one way or > another we should be able to visualize it in the tissues. > > Any ideas why the Candida-loaded positive control would not stain with > Grocott's? Right now I am thinking that perhaps they didn't sit long > enough in the 4% chromic acid, even though the protocol says to soak the > > slides for an hour to oxidize them. Either that or a longer soak in the > methenamine-silver solution at 60 degrees C (also for one hour). > > Thanks in advance for your help- > Kathleen Roberts > Principal Lab Technician > Neurotoxicology Labs > Dept of Pharmacology & Toxicology > Ernest Mario School of Pharmacy > Rutgers University > 41 B Gordon Rd > Piscataway, NJ 08854 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From kgrobert <@t> rci.rutgers.edu Wed May 5 14:18:52 2004 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Grocott's Methenamine Silver Stain In-Reply-To: References: Message-ID: <40993E1C.9020308@rci.rutgers.edu> I made up everything fresh just before I did the stain... :oP I'll try the 10% chromic acid. Thanks, Kathleen Wayne Holland wrote: > Make sure your solutions are fresh, especially the silver. ALso try > 10% chromic for 10 minutes. > > >> From: Kathleen Roberts >> To: "'histonet@lists.utsouthwestern.edu'" >> >> Subject: [Histonet] Grocott's Methenamine Silver Stain >> Date: Wed, 05 May 2004 13:54:18 -0400 >> >> Hey, all- >> >> I've been trying to stain some FFPE slides of decalcified rat sinuses >> for fungi using the AFIP protocol for Grocott's stain, and despite >> using fresh reagents, the positive control and unknowns do not stain >> at all, except for some random deposition of silver that does not >> even begin to look like fungi. The positive control is loaded with >> Candida, and can be seen with H&E easily. >> >> So far, we don't see any fungi in the rat sinuses and lungs (they >> were sneezing, and the bedding came back positive for Aspergillus) >> with H&E staining, which is why we decided to try the Grocott's-one >> way or another we should be able to visualize it in the tissues. >> >> Any ideas why the Candida-loaded positive control would not stain >> with Grocott's? Right now I am thinking that perhaps they didn't sit >> long enough in the 4% chromic acid, even though the protocol says to >> soak the slides for an hour to oxidize them. Either that or a longer >> soak in the methenamine-silver solution at 60 degrees C (also for one >> hour). >> >> Thanks in advance for your help- >> Kathleen Roberts >> Principal Lab Technician >> Neurotoxicology Labs >> Dept of Pharmacology & Toxicology >> Ernest Mario School of Pharmacy >> Rutgers University >> 41 B Gordon Rd >> Piscataway, NJ 08854 >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _________________________________________________________________ > Stop worrying about overloading your inbox - get MSN Hotmail Extra > Storage! > http://join.msn.com/?pgmarket=en-us&page=hotmail/es2&ST=1/go/onm00200362ave/direct/01/ > > From Sue.Kapoor <@t> uhsi.org Wed May 5 14:08:25 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Reichert Jung 2030 Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F55CB@khmcexch.uhsi.org> Does anyone know if there is a way to convert from high profile blades to low profile on the Reichert Jung 2030 microtome? Thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From histology.bc <@t> shaw.ca Wed May 5 14:32:09 2004 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Re: Polyester wax (with earlier citations) In-Reply-To: References: Message-ID: <40994139.8040303@shaw.ca> Hi Steve, Your descriptions bring back a flood of memories ... the wax tea pot, Leukhart's embedding rings, sticking the blocks onto wooden blocks, taking them off again at the end of the day, etc. Safety precautions had not even been invented in those days. When I first started in Histology, there were five of us in the lab and every single one smoked a pipe. So, embedding, trimming and sticking on the blocks involved three of us ... all chuffing out smoke. The conversations that took place during these times were priceless. Sadly, this opportunity was lost with the advent of automated embedding centres.. All solvents went down the drain, old specimens were dumped into the sink to allow the formalin to drain away. There was no fume hood, so the formalin fumes were thick enough to cut with a knife. In retrospect, dumping solvents and fixatives down the drain was not the best idea!... but at the time, that was standard practice. However, despite these horrendous practices, we are all still alive and well, and all went on to accept senior positions around the world.. There were no productivity units to count, no QC/QA demands (apart from self-imposed ones), no intrusions from mis-guided administrators. We had time to work on our own projects, investigate new procedures, and read journals looking for new methods. We made all of our own reagents from scratch (hematoxylin, Schiff's reagent, fixatives, etc) We sharpened our own knives. The camaraderie was wonderful, there was no bitching or whining, going for a beer at lunchtime was a routine practice. We did a great job, we went home happy, and provided great service I would not give up the new developments in Histology (immunohistochemistry, monoclonal antibodies, disposable knives, or automated stainers, etc ) they have produced quantum leaps in quality and diagnostic accuracy, but I sometimes I despair that the new generation of technologists have missed out on an invaluable learning experience. I firmly believe that I am a better histologist from my experience ... if something didn't work there was no service rep to call for advice, it was up to us to figure it out. The most respected "mentors" on the Histonet (who I won't name to avoid embarrasing them, but you know who I mean) are all the product of this bygone age of Histology. The Histonet serves a great purpose as a knowledge base and resource for advice, but there is no substitute for self-motivated learning. The books and journals are all out there ... waiting. Okay, I am done. Time to get down off my soapbox. The sermon is ended, Paul Bradbury Kamloops, BC Canada (formerly Nottingham, England) From kgrobert <@t> rci.rutgers.edu Wed May 5 14:43:42 2004 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Grocott's Methenamine Silver Stain In-Reply-To: <3.0.6.32.20040505131432.00c004a0@gemini.msu.montana.edu> References: <3.0.6.32.20040505131432.00c004a0@gemini.msu.montana.edu> Message-ID: <409943EE.8050800@rci.rutgers.edu> As for the decal, I was afraid that the decal solution (Fisher's Cal-EX) might have something to do with it...but the control block was tissue that did not need to be decal'd, so I think something else is afoot, and the decal isn't the whole story. True, about there possibly being no fungi in the lungs...these rats were sneezing, not coughing or wheezing, so I am told, so it may not have had time to spread to the lungs, but the vet wanted to check. H&E of some of the lungs does show an inflammatory reaction, however, even though there appear to be no fungi. Little to no inflammation seen in the sinuses, though that may have been eaten away by the decal, possibly. As for waterbath vs. incubator....I used an incubator. I'll try the waterbath next time. And I agree with you about comparing Candida control with Aspergillus...but this block is all we have right now, and the fact that *nothing is staining at all* with Grocott's, neither Candida control nor Aspergillus-infected subjects is fishy. Mmm, should I preheat the methenamine silver solution to 60 degrees C before putting the slides in? My protocol didn't say to do that, but I think someone here on the list said they did that...must search the archives. Thanks for your input! Kathleen Gayle Callis wrote: >It may be that your decalcification (if you used a strong acid and not >controlled with endpoint determination) may have messed up the stain. > >EDTA may have been a better choice for decalcification. However, you don't >decalcify lungs, and if you can't see the fungus in lung, Aspergillus is so >easy to stain with GMS, then there may not be any fungus there. Candida is >always a snap to stain, 1 hour in FRESH 4% chromic acid is standard. GMS >with Candida and Aspergillus usually comes up within or before 1 hour. You >should use a microscope to control silver deposition in positive Candida >staining. Are you using a waterbath or incubator? Waterbath is far better >for temperature control, more even distribution of heat, incubators have >convectional air currents. To check staining, dip in cold water, examine >then dip in hot distilled water to equilibrate section back to methenamine >silver temperature. If you don't dip in cold water, you get steam on your >microscope objectives. Look for that dark golden brown color at around 30 >min before pulling slide, then check every 15 minutes. > >Caveat: beware comparing positive Candida staining to your Aspergillus as >these ARE different fungi, Candida IS different and really stains readily. >Aspergillus may still be in yeast form phase of infection and not have >mycelium (a = plural) formation at point you euthanized rat. Seeing >Aspergillus readily in H&E is not always the case as with a heavy infection >of Candida, so be careful how you interpret these two in same breath. > >Good luck > >At 01:54 PM 5/5/2004 -0400, you wrote: > > >>Hey, all- >> >>I've been trying to stain some FFPE slides of decalcified rat sinuses >>for fungi using the AFIP protocol for Grocott's stain, and despite using >>fresh reagents, the positive control and unknowns do not stain at all, >>except for some random deposition of silver that does not even begin to >>look like fungi. The positive control is loaded with Candida, and can >>be seen with H&E easily. >> >>So far, we don't see any fungi in the rat sinuses and lungs (they were >>sneezing, and the bedding came back positive for Aspergillus) with H&E >>staining, which is why we decided to try the Grocott's-one way or >>another we should be able to visualize it in the tissues. >> >>Any ideas why the Candida-loaded positive control would not stain with >>Grocott's? Right now I am thinking that perhaps they didn't sit long >>enough in the 4% chromic acid, even though the protocol says to soak the >>slides for an hour to oxidize them. Either that or a longer soak in the >>methenamine-silver solution at 60 degrees C (also for one hour). >> >>Thanks in advance for your help- >>Kathleen Roberts >>Principal Lab Technician >>Neurotoxicology Labs >>Dept of Pharmacology & Toxicology >>Ernest Mario School of Pharmacy >>Rutgers University >>41 B Gordon Rd >>Piscataway, NJ 08854 >> >> >>_______________________________________________ >>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 kgrobert <@t> rci.rutgers.edu Wed May 5 14:46:38 2004 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Grocott's Methenamine Silver Stain In-Reply-To: <490C1EC04BA10F4891494D0ED756AC93223BF0@usmi0012k03.rallansci.com> References: <490C1EC04BA10F4891494D0ED756AC93223BF0@usmi0012k03.rallansci.com> Message-ID: <4099449E.1050905@rci.rutgers.edu> Aha! I knew there was a way to do that, but I could not find it in the archives. Thank you, I'll try it. -Kathleen Jones, Sarah - RAS wrote: >Kathleen, > >Since you don't know how long they were in the decal solution, you might >also try putting the slides (following deparaffinization and hydration and >before staining) into a saturated lithium carbonate solution for half an >hour or longer. This helps restore nuclear detail in over-decalcified >specimens, and might also help with your problem. > >Sarah > >-----Original Message----- >From: Kathleen Roberts [mailto:kgrobert@rci.rutgers.edu] >Sent: Wednesday, May 05, 2004 3:16 PM >To: Bartlett, Jeanine; 'histonet@lists.utsouthwestern.edu' >Subject: Re: [Histonet] Grocott's Methenamine Silver Stain > >I am not sure how long they were in decal...but I supplied some of our >Fisher Scientific Cal-EX, which has hydrochloric acid and EDTA in it, to >the Rutgers vet who had this case of sneezing rats. I did ask him how >long he decal'd them, but he doesn't remember either. > >The chromic acid was freshly ordered as a 10% solution from LabChem, >Inc., which was then diluted down to 4% the morning I was to use it. > >Thanks for your help! >Kathleen > >Bartlett, Jeanine wrote: > > > >>Kathleen: >> >>How long were they in decal and what type of decal? And do you make >>your chromic fresh? >> >>Jeanine Bartlett, HT(ASCP) >>Centers for Disease Control >>Infectious Disease Pathology Activity >>1600 Clifton Road, MS/G-32 >>Atlanta, GA 30333 >> >> >>-----Original Message----- >>From: histonet-bounces@lists.utsouthwestern.edu >>[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kathleen >>Roberts >>Sent: Wednesday, May 05, 2004 1:54 PM >>To: 'histonet@lists.utsouthwestern.edu' >>Subject: [Histonet] Grocott's Methenamine Silver Stain >> >> >>Hey, all- >> >>I've been trying to stain some FFPE slides of decalcified rat sinuses >>for fungi using the AFIP protocol for Grocott's stain, and despite using >> >>fresh reagents, the positive control and unknowns do not stain at all, >>except for some random deposition of silver that does not even begin to >>look like fungi. The positive control is loaded with Candida, and can >>be seen with H&E easily. >> >>So far, we don't see any fungi in the rat sinuses and lungs (they were >>sneezing, and the bedding came back positive for Aspergillus) with H&E >>staining, which is why we decided to try the Grocott's-one way or >>another we should be able to visualize it in the tissues. >> >>Any ideas why the Candida-loaded positive control would not stain with >>Grocott's? Right now I am thinking that perhaps they didn't sit long >>enough in the 4% chromic acid, even though the protocol says to soak the >> >>slides for an hour to oxidize them. Either that or a longer soak in the >>methenamine-silver solution at 60 degrees C (also for one hour). >> >>Thanks in advance for your help- >>Kathleen Roberts >>Principal Lab Technician >>Neurotoxicology Labs >>Dept of Pharmacology & Toxicology >>Ernest Mario School of Pharmacy >>Rutgers University >>41 B Gordon Rd >>Piscataway, NJ 08854 >> >> >>_______________________________________________ >>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 Wed May 5 14:41:42 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] PLP defintion In-Reply-To: <4A4E1F05-9ECB-11D8-A5EE-000393967904@utmem.edu> References: <3.0.6.32.20040505095456.00c015d8@gemini.msu.montana.edu> Message-ID: <3.0.6.32.20040505134142.00c004a0@gemini.msu.montana.edu> Sorry folks, PLP stands for Periodate- lysine-paraformaldehyde, McKean et al. A fixative sometimes used for immunostaining, particularly for CD markers. It can be perfused. 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 pam <@t> ategra.com Wed May 5 14:52:41 2004 From: pam <@t> ategra.com (Pam Barker (extension 234)) Date: Fri Sep 16 15:22:54 2005 Subject: [Histonet] Histo Techs and Histology Supervisors - Great opportunities with a New Lab in Florida !!! Message-ID: Histonetters - I am presently on a search for my newest client in the Tampa Bay Area who is seeking to hire a histology supervisor and histo techs. This is a BRAND NEW LAB. Work in a small cozy friendly environment with state of the art equipment. The hours are MON-FRI ONLY - D A Y S ONLY. (no working on weekends or holidays ! ) 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 231 EMAIL: pam@ategra.com WEBSITE: http://www.ategra.com From kgrobert <@t> rci.rutgers.edu Wed May 5 14:58:28 2004 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Grocott's Methenamine Silver Stain In-Reply-To: <3.0.6.32.20040505133710.00c004a0@gemini.msu.montana.edu> References: <3.0.6.32.20040505133710.00c004a0@gemini.msu.montana.edu> Message-ID: <40994764.5040703@rci.rutgers.edu> One thing-this wasn't a kit that I used, and I *am* following the protocol in the AFIP manual. The chromic acid is a 10% solution from LabChem, and I diluted it down to 4%. I suppose that they could have made a mistake in making the solution. How do you make up your chromic acid? Thanks for your help- Kathleen Gayle Callis wrote: >I don't thing 10% chromic is the answer, the method calls for 4%. Maybe >they made a mistake making it? Is it actually stronger than they say? > >You don't want to have Chromic too concentrated as if you overoxidize you >will take the vicinal OH groups to aldehyde and then beyond and get zip for >results. I have never used ready to use kits, and make up my 4% chromic >acid fresh each time. It is possible to OVEROXIDIZE with this method. > >A good read on this method is AFIP manual, Lee Luna tells HOW it should be >done very clearly. > > > >At 03:18 PM 5/5/2004 -0400, you wrote: > > >>I made up everything fresh just before I did the stain... :oP I'll >>try the 10% chromic acid. >> >>Thanks, >>Kathleen >> >>Wayne Holland wrote: >> >> >> >>>Make sure your solutions are fresh, especially the silver. ALso try >>>10% chromic for 10 minutes. >>> >>> >>> >>> >>>>From: Kathleen Roberts >>>>To: "'histonet@lists.utsouthwestern.edu'" >>>> >>>>Subject: [Histonet] Grocott's Methenamine Silver Stain >>>>Date: Wed, 05 May 2004 13:54:18 -0400 >>>> >>>>Hey, all- >>>> >>>>I've been trying to stain some FFPE slides of decalcified rat sinuses >>>>for fungi using the AFIP protocol for Grocott's stain, and despite >>>>using fresh reagents, the positive control and unknowns do not stain >>>>at all, except for some random deposition of silver that does not >>>>even begin to look like fungi. The positive control is loaded with >>>>Candida, and can be seen with H&E easily. >>>> >>>>So far, we don't see any fungi in the rat sinuses and lungs (they >>>>were sneezing, and the bedding came back positive for Aspergillus) >>>>with H&E staining, which is why we decided to try the Grocott's-one >>>>way or another we should be able to visualize it in the tissues. >>>> >>>>Any ideas why the Candida-loaded positive control would not stain >>>>with Grocott's? Right now I am thinking that perhaps they didn't sit >>>>long enough in the 4% chromic acid, even though the protocol says to >>>>soak the slides for an hour to oxidize them. Either that or a longer >>>>soak in the methenamine-silver solution at 60 degrees C (also for one >>>>hour). >>>> >>>>Thanks in advance for your help- >>>>Kathleen Roberts >>>>Principal Lab Technician >>>>Neurotoxicology Labs >>>>Dept of Pharmacology & Toxicology >>>>Ernest Mario School of Pharmacy >>>>Rutgers University >>>>41 B Gordon Rd >>>>Piscataway, NJ 08854 >>>> >>>> >>>>_______________________________________________ >>>>Histonet mailing list >>>>Histonet@lists.utsouthwestern.edu >>>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>>> >>>> >>>_________________________________________________________________ >>>Stop worrying about overloading your inbox - get MSN Hotmail Extra >>>Storage! >>> >>> >>> >http://join.msn.com/?pgmarket=en-us&page=hotmail/es2&ST=1/go/onm00200362ave/ >direct/01/ > > >>> >>> >>_______________________________________________ >>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 Don.Birgerson <@t> leica-microsystems.com Wed May 5 14:58:18 2004 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Reichert Jung 2030 Message-ID: Hi Sue, You can convert from Hi to Low or vise versa. The key is to identify which of the five possible holders "E' holders you have on your RM2030 so the correct part is ordered. Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Kapoor, Sue" To: histonet@pathology.swmed.edu Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] Reichert Jung 2030 western.edu 05/05/2004 02:08 PM Does anyone know if there is a way to convert from high profile blades to low profile on the Reichert Jung 2030 microtome? Thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> colobio.com Wed May 5 14:59:25 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] lymph nodes In-Reply-To: <7325D637DFE2D211928800902733A7F303B94ACC@DGAMTBDCEMS> Message-ID: I have for years used single edged razor blades to dissect unfixed tissues. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Mike Bromley Sent: Wednesday, May 05, 2004 9:26 AM To: Histonet (E-mail) Subject: [Histonet] lymph nodes Hi Everyone I am after a very thin blade to slice unfixed lymph nodes, does anyone have any ideas? Ideally some kind of holder for old style razor blades would be the answer. Best Wishes Mike Bromley Chief Biomedical Scientist Pathology Dumfries & Galloway Royal Infirmary Scotland, UK _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Wed May 5 15:03:13 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Grocott's Methenamine Silver Stain Message-ID: In an attempt to remove hazardous chromic acid from the lab, I vaguely remember replacing the Chromic Acid with 5% Periodic acid instead for the GMS. Has anyone else done this? If my memory serves me correctly - it works fine - but I am old . . . . er. (must be from all the teenagers at home). Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotherapeutics Kathleen Roberts , Sent by: "'histonet@lists.utsouthwestern.edu'" histonet-bounces@lists. utsouthwestern.edu cc: Subject: Re: [Histonet] Grocott's Methenamine Silver Stain 05/05/2004 02:58 PM One thing-this wasn't a kit that I used, and I *am* following the protocol in the AFIP manual. The chromic acid is a 10% solution from LabChem, and I diluted it down to 4%. I suppose that they could have made a mistake in making the solution. How do you make up your chromic acid? Thanks for your help- Kathleen Gayle Callis wrote: >I don't thing 10% chromic is the answer, the method calls for 4%. Maybe >they made a mistake making it? Is it actually stronger than they say? > >You don't want to have Chromic too concentrated as if you overoxidize you >will take the vicinal OH groups to aldehyde and then beyond and get zip for >results. I have never used ready to use kits, and make up my 4% chromic >acid fresh each time. It is possible to OVEROXIDIZE with this method. > >A good read on this method is AFIP manual, Lee Luna tells HOW it should be >done very clearly. > > > >At 03:18 PM 5/5/2004 -0400, you wrote: > > >>I made up everything fresh just before I did the stain... :oP I'll >>try the 10% chromic acid. >> >>Thanks, >>Kathleen >> >>Wayne Holland wrote: >> >> >> >>>Make sure your solutions are fresh, especially the silver. ALso try >>>10% chromic for 10 minutes. >>> >>> >>> >>> >>>>From: Kathleen Roberts >>>>To: "'histonet@lists.utsouthwestern.edu'" >>>> >>>>Subject: [Histonet] Grocott's Methenamine Silver Stain >>>>Date: Wed, 05 May 2004 13:54:18 -0400 >>>> >>>>Hey, all- >>>> >>>>I've been trying to stain some FFPE slides of decalcified rat sinuses >>>>for fungi using the AFIP protocol for Grocott's stain, and despite >>>>using fresh reagents, the positive control and unknowns do not stain >>>>at all, except for some random deposition of silver that does not >>>>even begin to look like fungi. The positive control is loaded with >>>>Candida, and can be seen with H&E easily. >>>> >>>>So far, we don't see any fungi in the rat sinuses and lungs (they >>>>were sneezing, and the bedding came back positive for Aspergillus) >>>>with H&E staining, which is why we decided to try the Grocott's-one >>>>way or another we should be able to visualize it in the tissues. >>>> >>>>Any ideas why the Candida-loaded positive control would not stain >>>>with Grocott's? Right now I am thinking that perhaps they didn't sit >>>>long enough in the 4% chromic acid, even though the protocol says to >>>>soak the slides for an hour to oxidize them. Either that or a longer >>>>soak in the methenamine-silver solution at 60 degrees C (also for one >>>>hour). >>>> >>>>Thanks in advance for your help- >>>>Kathleen Roberts >>>>Principal Lab Technician >>>>Neurotoxicology Labs >>>>Dept of Pharmacology & Toxicology >>>>Ernest Mario School of Pharmacy >>>>Rutgers University >>>>41 B Gordon Rd >>>>Piscataway, NJ 08854 >>>> >>>> >>>>_______________________________________________ >>>>Histonet mailing list >>>>Histonet@lists.utsouthwestern.edu >>>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>>> >>>> >>>_________________________________________________________________ >>>Stop worrying about overloading your inbox - get MSN Hotmail Extra >>>Storage! >>> >>> >>> >http://join.msn.com/?pgmarket=en-us&page=hotmail/es2&ST=1/go/onm00200362ave/ >direct/01/ > > >>> >>> >>_______________________________________________ >>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 carl.hobbs <@t> kcl.ac.uk Wed May 5 15:07:06 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] re S100 Message-ID: <000c01c432dc$8a54e570$7bcc9a51@home> I don't recall collagen positivity but certainly, adipocytes are positive ( still don't know why, but it's a beautiful Ab reagent for them...I used the Dako polyclonal on human tissue) --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.668 / Virus Database: 430 - Release Date: 24/04/2004 From gcallis <@t> montana.edu Wed May 5 15:37:16 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Periodic acid versus chromic acid for GMS staining In-Reply-To: Message-ID: <3.0.6.32.20040505143716.00bfe0e8@gemini.msu.montana.edu> Caveat: Periodic acid may not be the acceptable oxidizing agent for fungus staining, there is a publication on this by Freida Carson with Jerry Fredenburgh in J of Histotechnology a few years back. Periodic acid is not strong enough to do the job i.e. oxidation on fungal components. One can get false negative results. A lot of people use this, but may not be aware of problems one can have doing GMS with periodic acid oxidation - it works fine for basement membrane staining in 2 um kidney sections i.e. Jones methenamine silver, but for accurate results, chromic acid may be the better choice for fungus work. There was a huge discussion on Histonet on this subject a few years back. Also, you still have to collect the silver solution as hazardous waste so one more thing should not be too much of a problem. I do feel for those who have to pay for this collection/waste disposal, we are fortunate our university picks up that cost. At 03:03 PM 5/5/2004 -0500, you wrote: > >In an attempt to remove hazardous chromic acid from the lab, I vaguely >remember replacing the Chromic Acid with 5% Periodic acid instead for the GMS. >Has anyone else done this? If my memory serves me correctly - it works fine - >but I am old . . . . er. (must be from all the teenagers at home). > >Jacqueline M. O'Connor HT(ASCP) >Abbott Laboratories >Global Pharmaceutical Research and Development >Discovery Chemotherapeutics > 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 kgrobert <@t> rci.rutgers.edu Wed May 5 15:47:12 2004 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Grocott's Methenamine Silver Stain In-Reply-To: <3.0.6.32.20040505142705.00bfe0e8@gemini.msu.montana.edu> References: <3.0.6.32.20040505133710.00c004a0@gemini.msu.montana.edu> <3.0.6.32.20040505133710.00c004a0@gemini.msu.montana.edu> <3.0.6.32.20040505142705.00bfe0e8@gemini.msu.montana.edu> Message-ID: <409952D0.3020603@rci.rutgers.edu> OK, so I guess I'll get some solid chromium trioxide and make it up that way and see if it makes any difference, though if the samples are over-oxidized, I may not be able to get anywhere with them...at least I'll be able to work it out with the control slides. Thanks for the reminder about disposal...you reminded me that, since I am the chemical safety officer for the lab, that I need to get a list together of all the stuff that Rutgers Env. Health & Safety needs to pick up and send it over to them. My to-do list never ends... :o) Thank you very much for your help- Kathleen Gayle Callis wrote: >Dissolve 4 gms chromium trioxide in 100 mls of distilled water, a simple >chemical solution. Chromic acid is a misnomer from years ago, it is made >from chromium trioxide. I have used 5% but 4% is the norm. I do use a >solution for 1 week if I am still doing the stain the next day. > >Remember you CANNOT dump chromium solutions down the drain, toxic waste and >must be collected. > >By the way, lithium carbonate will only neutralize any residual acid left >in tissue which damages nuclei by acid hydrolysis of DNA/RNA, and may help >restore the staining of hematoxyling. Howeer, LiCO3, a base, does NOT >reverse the chemical effects of acid IF you have overoxidized the groups >you want to bind silver to with GMS. Once protein hydrolysis is done and >gone beyond the endpoint you need, you are in trouble. > >This is the reason that hydrochloric acid and even formic acid is not >advised IF you want to do a Feulgens staining reaction for DNA, the >hydrolyzation step is basically overdone, and you get weak to no 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 froyer <@t> bitstream.net Wed May 5 14:51:43 2004 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Reichert Jung 2030 In-Reply-To: <61E9F2400F53D5119CFC00508B44E33B019F55CB@khmcexch.uhsi.org> References: <61E9F2400F53D5119CFC00508B44E33B019F55CB@khmcexch.uhsi.org> Message-ID: <409945CF.9000509@bitstream.net> Sue, It depends on which style of blade holder assembly you have. Older style... you need to replace the high profile back pressure plate with the low profile plate. Newer style... all you need is a slide-in bar that accepts the low profile blades. ~ Ford Ford M. Royer, MT(ASCP) Analytical Instruments, LLC Minneapolis, MN 55427 (800) 565-1895 Kapoor, Sue wrote: >Does anyone know if there is a way to convert from high profile blades to >low profile on the Reichert Jung 2030 microtome? > >Thanks in advance, >Sue Kapoor, HT (ASCP) >Histology Coordinator >Kenosha Medical Center >Kenosha, WI >262-653-5570 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From portera203 <@t> yahoo.com Wed May 5 15:41:50 2004 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Grocott's Methenamine Silver Stain In-Reply-To: <40992A4A.3000604@rci.rutgers.edu> Message-ID: <20040505204150.19445.qmail@web40908.mail.yahoo.com> I would be suspicious about the borax. We do decalcified mouse turbinates with this method for fungal sinusitis. Any time I have no or weak reaction i make up fresh borax. Since you are making everything up fresh, ?????. If your control hasn't been decaled then it seems like that should be working if all your solutions are fresh. I use a water-bath and prewarm my working silver before i add my slides. Hope you figure out what is wrong, very frustrating. Kathleen Roberts wrote: Hey, all- I've been trying to stain some FFPE slides of decalcified rat sinuses for fungi using the AFIP protocol for Grocott's stain, and despite using fresh reagents, the positive control and unknowns do not stain at all, except for some random deposition of silver that does not even begin to look like fungi. The positive control is loaded with Candida, and can be seen with H&E easily. So far, we don't see any fungi in the rat sinuses and lungs (they were sneezing, and the bedding came back positive for Aspergillus) with H&E staining, which is why we decided to try the Grocott's-one way or another we should be able to visualize it in the tissues. Any ideas why the Candida-loaded positive control would not stain with Grocott's? Right now I am thinking that perhaps they didn't sit long enough in the 4% chromic acid, even though the protocol says to soak the slides for an hour to oxidize them. Either that or a longer soak in the methenamine-silver solution at 60 degrees C (also for one hour). Thanks in advance for your help- Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Dept of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers University 41 B Gordon Rd Piscataway, NJ 08854 _______________________________________________ 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!? Win a $20,000 Career Makeover at Yahoo! HotJobs From convmcm <@t> cc.usu.edu Wed May 5 16:31:48 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Basics - IHC - CORRECTION In-Reply-To: <000001c432c3$b91f3c20$4a737b81@Cygnus> Message-ID: <000201c432e8$60caee50$4a737b81@Cygnus> OOOPSIE-DOOOPSIE! I made a mistake! The other Antibody publication I downloaded was from chemicon, NOT US Biological ... I must have visited too many websites per minute and got a bit of fuzz in my brain. That happens. For the introduction to antibodies,go to www.chemicon.com Sorry about that! Whilst I have your attention... I am in sore need of Bovine Parainfluenza 3 antibodies for IHC on ffpe. I also need bovine RSV, although I think US Biological has what I need. I'm waiting to hear back from them on this. can anyone point me to a vendor who carries this ab? Thanks so much, everyone! Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Connie McManus Sent: Wednesday, May 05, 2004 10:09 AM To: balajimr@drreddys.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Basics - IHC DAKO Cytomation has the best handbook in PDF format on IHC and it's free on the web. Got to www.dakocytomation.com. Also US Biological has a PDF publication called Introduction to antibodies that seems pretty good, too. I just downloaded it today, so I can't say what it's like, but I suppose it will be a good reference. Cheers. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of balajimr@drreddys.com Sent: Tuesday, May 04, 2004 8:26 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Basics - IHC Importance: High I have to prepare for an inhouse seminar on basics of immunohistochemistry and its aplications in research. Could anyone suggest me best/ideal literature/reference available on the web for the preparationof the same. Balaji _______________________________________________ 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 histology.bc <@t> shaw.ca Wed May 5 16:42:37 2004 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Grocott's Methenamine Silver Stain In-Reply-To: <409952D0.3020603@rci.rutgers.edu> References: <3.0.6.32.20040505133710.00c004a0@gemini.msu.montana.edu> <3.0.6.32.20040505133710.00c004a0@gemini.msu.montana.edu> <3.0.6.32.20040505142705.00bfe0e8@gemini.msu.montana.edu> <409952D0.3020603@rci.rutgers.edu> Message-ID: <40995FCD.2010405@shaw.ca> Hi Kathleen, Just a few thoughts about your lack of reaction with the GMS. Basically, what you are trying to show here is the neutral mucopolysaccharides in the capsule of the fungal elements. They wil stain intensely with a PAS, however, so will a whole bunch of other stuff (glyogen, contents of some goblet cells, basal lamina, etc), but fungal elements are morphologically quite easy to distinguish. The oxidation step in the PAS involves a relatively short treatment in 1% periodic acid, this will convert any available 1:2-glycols to aldehyde groups, which in turn react with the Schiff reagent ... consequently, a whole bunch of stuff reacts positively. The GMS will demonstrate only structures which are strongly PAS-positive. Instead of oxidizing in periodic acid, thr GMS uses chromic acid (a much more powerful oxidizer). Many of the weaker reactive groups over-oxidize and become non-reactive with the subsequent silver solution. Glycogen, neutral mucopolysaccharides in goblet cells, and fungi remain reactive ... and should react with the silver solution. The GMS is a pretty reliable procedure. Try using 10% chromic acid for 10 minutes (I developed this modification when I was teaching Histology and had to be able to perform the whole method within a two hour lab period). It works great and several labs have adopted it for use ... the whole method can be done in less than 30 minutes! The 10% chromic acid solution is stable for several months. How old is your stock methenamine-silver solution? Do you keep it in the fridge? If in doubt, make up some fresh and store it at 4 degrees C. The borax (sodium tetraborate) acts as a buffer the make the working silver solution slightly alkaline (and unstable). Mix the working solution as follows: 25mL of methenamine-silver stock solution, 25mL distilled water, 2mL borax. Preheat the silver solution to 56 degrees. Heat the solution just before you need it, not too soon or all the silver will precipitate out before it has chance to react with the fungal elements in the section. The silver impregnation step should require 6-10 minutes. You will see the section begin to turn :golden-light brown) as the silver reacts with the tissues. Before and after the silver step, rinse the sections with several changes of distilled water. Use plastic forceps to avoid getting any metallic ions into the silver solution. The method: 1. Take sections down to water 2 Oxidize in 10% chromic acid for 10 minutes 3 Wash in water for 1 minutes 4. Treat with 0.5-1% sodium bisulphite (gets rid of excess chromic acid) 5. Wash section in several changes of distilled water (cleanliness is a virtue) 6. Place sections in pre-warmed methenamine-silver working solution, keep temperature at 56 degrees. 7. Watch them like hawk ... within 4-5 minutes the sections will start to turn golden. 8. When they are light brown, take the control out, rinse in distilled water and look at it microscopically. Fungal hyphae should appear black on a light gold background. 9. If the fungi are still brown, return the section to the warm silver solution and give it another 30 seconds or so. Then check it again. 10. When you are happy with silver impregnation, remove all sections from the silver solution and wash thoroughly in distilled water. 11. Treat sections with 1-2% sodium thiosulphate for 1 minute. 12. Wash again 13. Treat sections with gold chloride (whatever strength you have on hand) for 1-2 minutes, 14. Wash yet again, 15. Counterstain with light green. 16. Wash (last time) 17. Dehydrate, clear and mount. Good luck, Paul Kamloops, BC, Canada From carl.hobbs <@t> kcl.ac.uk Wed May 5 16:55:15 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] re PLP fixing fluid Message-ID: <002f01c432eb$a5ec0bb0$7bcc9a51@home> Like lots of fixation exotics devised by many( mostly not Histotechs)....would be very grateful to receive feedback on them ie are they superior to PFA/formalin??? Not IMHO. Have tried many and not found anything superior to formalin. OK....Perfix/pwax, for eg is great for Histone3...but so is formalin/pwax/Ag heat retrieval and....one can utilise many other Ab reagents on the latter, unlike Perfix?or other exotica --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.668 / Virus Database: 430 - Release Date: 24/04/2004 From mrsgbd2001 <@t> yahoo.com Wed May 5 18:06:34 2004 From: mrsgbd2001 <@t> yahoo.com (Gareth Davis) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] cryo rat turbinates Message-ID: <20040505230634.49439.qmail@web13310.mail.yahoo.com> Okay, so much for my protocol for frozen rat turbinates. I tried to do an H&E on the turbinate sections I cut (in OCT and then cut and placed on charged slides). But once the sections hit the water rinses they all came off my slides. Anyone with suggestions? The sections were dried overnight at room temperature on the charged slides, and then stored at -80. Before staining I let them come to room temperature again. I then followed a H&E protocol I found on the Histonet, but, like I said, they all came off in the water rinses. So, if anyone can help me I would appreciate it. We were going to do IHC with antigen retrieval on these sections, but we decided not to even try that. Thanks, Gareth Davis Research Assistant Tennessee State University Nashville, Tennessee --------------------------------- Do you Yahoo!? Win a $20,000 Career Makeover at Yahoo! HotJobs From bhewlett <@t> cogeco.ca Wed May 5 19:11:27 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Re: Polyester wax (with earlier citations) References: <40994139.8040303@shaw.ca> Message-ID: <005601c432fe$ad10d520$6500a8c0@mainbox> Hi Paul, Lots of good times; but you forgot about the blistered finger tips from sticking those wax blocks on wooden blocks(the smaller the block-the bigger the blister!), and all the hand processing, which mainly involved much frantic dashing about twirling multiple pots of tissue to provide agitation! Who can forget about mouth pipetting whilst eating sandwiches at the staining bench and brewing the tea over a bunsen during waiting times! If I recall correctly, the pipes were full of either St. Bruno flake or 'Herbal' tobacco and between chuffs of smoke, most of the conversation was punctuated by phrases such as "eh up", "y'whot", "gerroff" and "owd ya tight", interspersed with "cool man", "crazy daddy" and "all gone". Ahhh... sweet memories. Bryan Hewlett Hamilton, Ont. Canada (formerly Nottingham, England) ----- Original Message ----- From: "Paul Bradbury" To: ; "HistoNet Server" Sent: Wednesday, May 05, 2004 3:32 PM Subject: Re: [Histonet] Re: Polyester wax (with earlier citations) > Hi Steve, > > Your descriptions bring back a flood of memories ... the wax tea pot, > Leukhart's embedding rings, sticking the blocks onto wooden blocks, > taking them off again at the end of the day, etc. Safety precautions had > not even been invented in those days. When I first started in Histology, > there were five of us in the lab and every single one smoked a pipe. So, > embedding, trimming and sticking on the blocks involved three of us ... > all chuffing out smoke. The conversations that took place during these > times were priceless. Sadly, this opportunity was lost with the advent > of automated embedding centres.. > > All solvents went down the drain, old specimens were dumped into the > sink to allow the formalin to drain away. There was no fume hood, so the > formalin fumes were thick enough to cut with a knife. In retrospect, > dumping solvents and fixatives down the drain was not the best idea!... > but at the time, that was standard practice. However, despite these > horrendous practices, we are all still alive and well, and all went on > to accept senior positions around the world.. > > There were no productivity units to count, no QC/QA demands (apart from > self-imposed ones), no intrusions from mis-guided administrators. We had > time to work on our own projects, investigate new procedures, and read > journals looking for new methods. We made all of our own reagents from > scratch (hematoxylin, Schiff's reagent, fixatives, etc) We sharpened our > own knives. The camaraderie was wonderful, there was no bitching or > whining, going for a beer at lunchtime was a routine practice. We did a > great job, we went home happy, and provided great service > > I would not give up the new developments in Histology > (immunohistochemistry, monoclonal antibodies, disposable knives, or > automated stainers, etc ) they have produced quantum leaps in quality > and diagnostic accuracy, but I sometimes I despair that the new > generation of technologists have missed out on an invaluable learning > experience. > > I firmly believe that I am a better histologist from my experience ... > if something didn't work there was no service rep to call for advice, it > was up to us to figure it out. The most respected "mentors" on the > Histonet (who I won't name to avoid embarrasing them, but you know who I > mean) are all the product of this bygone age of Histology. The Histonet > serves a great purpose as a knowledge base and resource for advice, but > there is no substitute for self-motivated learning. The books and > journals are all out there ... waiting. > > Okay, I am done. Time to get down off my soapbox. The sermon is ended, > > Paul Bradbury > Kamloops, BC > Canada > (formerly Nottingham, England) > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Stephen.Eyres <@t> sanofi-synthelabo.com Thu May 6 03:49:24 2004 From: Stephen.Eyres <@t> sanofi-synthelabo.com (Stephen.Eyres@sanofi-synthelabo.com) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Re: Polyester wax (with earlier citations) Message-ID: Hi Paul, I think you are absolutely right. The advancements have helped loads but the atmosphere and the 'family' spirit seems to have gone. Yo worked in Nottingham. I was in Leicester Royal, and attended the QMC for an advanced histology course in 1975 or 6 ( the effect of xylene demylination, I think), before the special courses began in earnest. Did you work with Neil hand? Cheers Steve Paul Bradbury HistoNet Server cc: 05/05/2004 20:32 Subject: Re: [Histonet] Re: Polyester wax (with earlier citations) Hi Steve, Your descriptions bring back a flood of memories ... the wax tea pot, Leukhart's embedding rings, sticking the blocks onto wooden blocks, taking them off again at the end of the day, etc. Safety precautions had not even been invented in those days. When I first started in Histology, there were five of us in the lab and every single one smoked a pipe. So, embedding, trimming and sticking on the blocks involved three of us ... all chuffing out smoke. The conversations that took place during these times were priceless. Sadly, this opportunity was lost with the advent of automated embedding centres.. All solvents went down the drain, old specimens were dumped into the sink to allow the formalin to drain away. There was no fume hood, so the formalin fumes were thick enough to cut with a knife. In retrospect, dumping solvents and fixatives down the drain was not the best idea!... but at the time, that was standard practice. However, despite these horrendous practices, we are all still alive and well, and all went on to accept senior positions around the world.. There were no productivity units to count, no QC/QA demands (apart from self-imposed ones), no intrusions from mis-guided administrators. We had time to work on our own projects, investigate new procedures, and read journals looking for new methods. We made all of our own reagents from scratch (hematoxylin, Schiff's reagent, fixatives, etc) We sharpened our own knives. The camaraderie was wonderful, there was no bitching or whining, going for a beer at lunchtime was a routine practice. We did a great job, we went home happy, and provided great service I would not give up the new developments in Histology (immunohistochemistry, monoclonal antibodies, disposable knives, or automated stainers, etc ) they have produced quantum leaps in quality and diagnostic accuracy, but I sometimes I despair that the new generation of technologists have missed out on an invaluable learning experience. I firmly believe that I am a better histologist from my experience ... if something didn't work there was no service rep to call for advice, it was up to us to figure it out. The most respected "mentors" on the Histonet (who I won't name to avoid embarrasing them, but you know who I mean) are all the product of this bygone age of Histology. The Histonet serves a great purpose as a knowledge base and resource for advice, but there is no substitute for self-motivated learning. The books and journals are all out there ... waiting. Okay, I am done. Time to get down off my soapbox. The sermon is ended, Paul Bradbury Kamloops, BC Canada (formerly Nottingham, England) From Stephen.Eyres <@t> sanofi-synthelabo.com Thu May 6 04:12:08 2004 From: Stephen.Eyres <@t> sanofi-synthelabo.com (Stephen.Eyres@sanofi-synthelabo.com) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Re: Polyester wax (with earlier citations) Message-ID: Hi Bryan, Paul. Did you ever put mince pie in the incubators at Christmas? We did and I had a very nasty surprise one year when I tasted one and found that someone had put a block for reprocessing into xylene in the incubator. No a pleaseant taste. I remember one very embarrasing incident. We had a pathologist who performed skin tests on her patients in her office. For a laugh, one guy brought a gorilla hand glove in, put it on and wrapped his hand in a towel and we rushed to her office, one knock, and I burst in and exclaimed..'Tess, quick look at Tony's hand he's hurt it' Looking round anxiously at the cover hand, Tony wipped the towel off and we burst out laughing. Tess looked passed us to the corner. There sat a confused and alarmed patient. We beat a hasty retreat, still laughing! Did you have a dress code? We had to wear shirt and tie-just in case we bumped into a patient in the corridor. Mind, that phased out in the early 80's, though it has stuck with me and I still wear a tie to this day. Did you ever experience the feeling of dread when you placed you beloved knife on the kemit plate only to see a a string of copper curl up onto the bevel? In the panic, people never managed to find the right button to turn it off. And the there was the furtive look to see if anyone had noticed. We bought a Temtool knife sharpener when they first came out. Arrrr, the fun we had with it. Trying to get the spirit level right, trying to buff the edge with out cutting the diamond strip. The patholgist i mentioned in a previous message, actually tried to sharpen his pen-knife in the thing, by holding it against the wheels. The bade got caught, was dragged out of his hand and had the blade snalled into 3 pieces. Miraculously, he wasn't injured. Cheers Steve "Bryan Hewlett" To: "Paul Bradbury" Sent by: Stephen Eyres/GB-ALNWICK/RESEARCH/SANOFI@Research histonet-bounces@lists.utsouth "HistoNet Server" western.edu cc: Subject: Re: [Histonet] Re: Polyester wax (with earlier citations) 06/05/2004 01:11 Hi Paul, Lots of good times; but you forgot about the blistered finger tips from sticking those wax blocks on wooden blocks(the smaller the block-the bigger the blister!), and all the hand processing, which mainly involved much frantic dashing about twirling multiple pots of tissue to provide agitation! Who can forget about mouth pipetting whilst eating sandwiches at the staining bench and brewing the tea over a bunsen during waiting times! If I recall correctly, the pipes were full of either St. Bruno flake or 'Herbal' tobacco and between chuffs of smoke, most of the conversation was punctuated by phrases such as "eh up", "y'whot", "gerroff" and "owd ya tight", interspersed with "cool man", "crazy daddy" and "all gone". Ahhh... sweet memories. Bryan Hewlett Hamilton, Ont. Canada (formerly Nottingham, England) ----- Original Message ----- From: "Paul Bradbury" To: ; "HistoNet Server" Sent: Wednesday, May 05, 2004 3:32 PM Subject: Re: [Histonet] Re: Polyester wax (with earlier citations) > Hi Steve, > > Your descriptions bring back a flood of memories ... the wax tea pot, > Leukhart's embedding rings, sticking the blocks onto wooden blocks, > taking them off again at the end of the day, etc. Safety precautions had > not even been invented in those days. When I first started in Histology, > there were five of us in the lab and every single one smoked a pipe. So, > embedding, trimming and sticking on the blocks involved three of us ... > all chuffing out smoke. The conversations that took place during these > times were priceless. Sadly, this opportunity was lost with the advent > of automated embedding centres.. > > All solvents went down the drain, old specimens were dumped into the > sink to allow the formalin to drain away. There was no fume hood, so the > formalin fumes were thick enough to cut with a knife. In retrospect, > dumping solvents and fixatives down the drain was not the best idea!... > but at the time, that was standard practice. However, despite these > horrendous practices, we are all still alive and well, and all went on > to accept senior positions around the world.. > > There were no productivity units to count, no QC/QA demands (apart from > self-imposed ones), no intrusions from mis-guided administrators. We had > time to work on our own projects, investigate new procedures, and read > journals looking for new methods. We made all of our own reagents from > scratch (hematoxylin, Schiff's reagent, fixatives, etc) We sharpened our > own knives. The camaraderie was wonderful, there was no bitching or > whining, going for a beer at lunchtime was a routine practice. We did a > great job, we went home happy, and provided great service > > I would not give up the new developments in Histology > (immunohistochemistry, monoclonal antibodies, disposable knives, or > automated stainers, etc ) they have produced quantum leaps in quality > and diagnostic accuracy, but I sometimes I despair that the new > generation of technologists have missed out on an invaluable learning > experience. > > I firmly believe that I am a better histologist from my experience ... > if something didn't work there was no service rep to call for advice, it > was up to us to figure it out. The most respected "mentors" on the > Histonet (who I won't name to avoid embarrasing them, but you know who I > mean) are all the product of this bygone age of Histology. The Histonet > serves a great purpose as a knowledge base and resource for advice, but > there is no substitute for self-motivated learning. The books and > journals are all out there ... waiting. > > Okay, I am done. Time to get down off my soapbox. The sermon is ended, > > Paul Bradbury > Kamloops, BC > Canada > (formerly Nottingham, England) > > > > > > > > > _______________________________________________ > 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 Myri37 <@t> aol.com Thu May 6 04:59:49 2004 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] cryoprotection before MMA embedding at -20 =?iso-8859-1?q?=B0C?= Message-ID: <71FE4E26.5FC3474F.0005167B@aol.com> Hi everyone i use a protocol for MMA embedding at 20?C since a few months. and this protocol says to put simples in phosphate buffer 7.4 with 10% sucrose overnight at 4?C after fixation in paraformald?hyde 4% for 3 days and before dehydratation and preimpregnation at 4?C, emebdding is done at -20?C. Last time i forget the step of cryoprotection with sucrose, and after cutting and staining my embedding simples, i didn't see any cells, no one cell !!! i did never see that before !!!! Do you think that is because of no cryoprotection ? Does a lack cryoprotection induce disparition of cells ? the tissue was very thin... any help would be very usefull Thank you very much Myriam Baali Natural implant From kemlo <@t> tiscali.co.uk Thu May 6 05:20:49 2004 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] lymph nodes In-Reply-To: Message-ID: <000501c43353$d1727d60$02dde150@KEMLOS> Yes I agree, if you want to save money, and you are in Scotland, you can use those discarded from cutting sections; as long as it wasn't bone! Mr Kemlo Rogerson MSc DMS MIBiol Cbiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 05 May 2004 16:37 To: Mike Bromley; Histonet (E-mail) Subject: RE: [Histonet] lymph nodes I find for this sort of thing, disposable microtome blades are ideal. Holders are available, but I neither use nor recommend them. 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: Mike Bromley [mailto:M.Bromley@dgri.scot.nhs.uk] Sent: 05 May 2004 16:26 To: Histonet (E-mail) Subject: [Histonet] lymph nodes Hi Everyone I am after a very thin blade to slice unfixed lymph nodes, does anyone have any ideas? Ideally some kind of holder for old style razor blades would be the answer. Best Wishes Mike Bromley Chief Biomedical Scientist Pathology Dumfries & Galloway Royal Infirmary Scotland, 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 ian.montgomery <@t> bio.gla.ac.uk Thu May 6 05:54:47 2004 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:22:55 2005 Subject: Fwd: Re: [Histonet] Re: Polyester wax (with earlier citations) Message-ID: <6.0.1.1.2.20040506113746.03318460@udcf.gla.ac.uk> Here I am in sunny Glasgow living in a time warp. Automation, what's that? Hand process, make my own stains, sharpen knives, name it and I'm still doing it. Well apart from wooden blocks, gave that up 30 odd years ago. I'm such a sad case that if you ask me what a particular chemical in my store is for I'll tell you exactly and how much is used. Sad thing is, I'll be retiring shortly and all the knowledge will go with me. Ian. >Hi Paul, > >Lots of good times; >but you forgot about the blistered finger tips from sticking those wax >blocks on wooden blocks(the smaller the block-the bigger the blister!), and >all the hand processing, which mainly involved much frantic dashing about >twirling multiple pots of tissue to provide agitation! >Who can forget about mouth pipetting whilst eating sandwiches at the >staining bench and brewing the tea over a bunsen during waiting times! >If I recall correctly, the pipes were full of either St. Bruno flake or >'Herbal' tobacco and between chuffs of smoke, most of the conversation was >punctuated by phrases such as "eh up", "y'whot", "gerroff" and "owd ya >tight", interspersed with "cool man", "crazy daddy" and "all gone". Ahhh... >sweet memories. > >Bryan Hewlett >Hamilton, Ont. >Canada >(formerly Nottingham, England) > >----- Original Message ----- >From: "Paul Bradbury" >To: ; "HistoNet Server" > >Sent: Wednesday, May 05, 2004 3:32 PM >Subject: Re: [Histonet] Re: Polyester wax (with earlier citations) > > > > Hi Steve, > > > > Your descriptions bring back a flood of memories ... the wax tea pot, > > Leukhart's embedding rings, sticking the blocks onto wooden blocks, > > taking them off again at the end of the day, etc. Safety precautions had > > not even been invented in those days. When I first started in Histology, > > there were five of us in the lab and every single one smoked a pipe. So, > > embedding, trimming and sticking on the blocks involved three of us ... > > all chuffing out smoke. The conversations that took place during these > > times were priceless. Sadly, this opportunity was lost with the advent > > of automated embedding centres.. > > > > All solvents went down the drain, old specimens were dumped into the > > sink to allow the formalin to drain away. There was no fume hood, so the > > formalin fumes were thick enough to cut with a knife. In retrospect, > > dumping solvents and fixatives down the drain was not the best idea!... > > but at the time, that was standard practice. However, despite these > > horrendous practices, we are all still alive and well, and all went on > > to accept senior positions around the world.. > > > > There were no productivity units to count, no QC/QA demands (apart from > > self-imposed ones), no intrusions from mis-guided administrators. We had > > time to work on our own projects, investigate new procedures, and read > > journals looking for new methods. We made all of our own reagents from > > scratch (hematoxylin, Schiff's reagent, fixatives, etc) We sharpened our > > own knives. The camaraderie was wonderful, there was no bitching or > > whining, going for a beer at lunchtime was a routine practice. We did a > > great job, we went home happy, and provided great service > > > > I would not give up the new developments in Histology > > (immunohistochemistry, monoclonal antibodies, disposable knives, or > > automated stainers, etc ) they have produced quantum leaps in quality > > and diagnostic accuracy, but I sometimes I despair that the new > > generation of technologists have missed out on an invaluable learning > > experience. > > > > I firmly believe that I am a better histologist from my experience ... > > if something didn't work there was no service rep to call for advice, it > > was up to us to figure it out. The most respected "mentors" on the > > Histonet (who I won't name to avoid embarrasing them, but you know who I > > mean) are all the product of this bygone age of Histology. The Histonet > > serves a great purpose as a knowledge base and resource for advice, but > > there is no substitute for self-motivated learning. The books and > > journals are all out there ... waiting. > > > > Okay, I am done. Time to get down off my soapbox. The sermon is ended, > > > > Paul Bradbury > > Kamloops, BC > > Canada > > (formerly Nottingham, England) > > > > > > > > > > > > > > > > > > _______________________________________________ > > 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 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 c.m.vanderloos <@t> amc.uva.nl Thu May 6 05:55:40 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] RE: double immuno staining Message-ID: <5fccd95f89ae.5f89ae5fccd9@amc.uva.nl> Dear Veronique, I am wondering if it would be possible to embed the specimens after the perfusion into paraffin. This would make your tissue morphology more "resistant" to pretreatment procedures. Since you have it so optimally fixed by perfusion one may wonder what can go wrong with the antigens during full paraffin embedding? I am a bit surprised about killing the staining of desmin. Usually this is a good and stable antigen that cannot leak away or so. Perhaps testing different clones of anti-desmin is helpful to you. Hope this helps, Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From "Veronique Andriessen" Date Wed, 5 May 2004 16:19:08 +0200 To "Histonet" Subject [Histonet] double immuno staining Hi folks, I need some help. I am struggling with a double immunostaining for somatostatin receptor and desmin. All my previous double stainings worked beautifully, but they were performed on fresh frozen acetone fixed pancreatic tissue. For staining of somatostatin receptor I have to fix with formaldehyde. I use 20 minutes 0.75% PFA perfused tissue which is snap frozen embedded in tissue freezing medium and cut on a cryostat. I was advised to do it this way to prevent loss of antigenicity. Morphology is quite good. This fixation works for the somatostatin receptor staining, but it kills the desmin signal. I was able to retrieve most of the desmin signal by EIER for 10 minutes with trypsin-EDTA, but now the morphology is awful. Can someone give me some advice? Sincerely yours, Veronique Andriessen BAS Lab. Molecular Liver Cell Biology, Free University Brussels (VUB)Belgium From TMcNemar <@t> lmhealth.org Thu May 6 06:21:38 2004 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Lab Storage - floor pallettes Message-ID: <90092A4ED388D7119575006008F7112049CB95@NT_EXCHANGE> We just went through that last year. We had maintenance make them up for us. All they did was attach 2x2s to a piece of melamine. Any size we wanted. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Cheasty, Sandra [mailto:SCheasty@ahs.llumc.edu] Sent: Wednesday, May 05, 2004 10:58 AM To: HistoNet (E-mail) Subject: [Histonet] Lab Storage - floor pallettes Does anyone out there have a source for floor palettes that are only 24" deep? The only ones I can find so far are all 40" deep. (We need to store some supplies off the floor under the counters.) Thank you, Sandy Confidentiality Note: The preceding e-mail message (including any attachments) contains information that may be confidential, protected by applicable legal privileges, or constitute non-public information. It is intended to be conveyed only to the designated recipient(s). If you are not an intended recipient of this message, please notify the sender by replying to this message and then delete it from your system. Use, dissemination, distribution or reproduction of this message by unintended recipients is not authorized and may be unlawful. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kgrobert <@t> rci.rutgers.edu Thu May 6 08:52:42 2004 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Periodic acid versus chromic acid for GMS staining In-Reply-To: <3.0.6.32.20040505143716.00bfe0e8@gemini.msu.montana.edu> References: <3.0.6.32.20040505143716.00bfe0e8@gemini.msu.montana.edu> Message-ID: <409A432A.8020701@rci.rutgers.edu> Interesting, because Richard-Allan Scientific has a kit that apparently uses 1% periodic acid... http://www.rallansci.com/histology/histology.aspx?id=61 Still scratching my head today, Kathleen Gayle Callis wrote: >Caveat: Periodic acid may not be the acceptable oxidizing agent for fungus >staining, there is a publication on this by Freida Carson with Jerry >Fredenburgh in J of Histotechnology a few years back. Periodic acid is not >strong enough to do the job i.e. oxidation on fungal components. One can >get false negative results. A lot of people use this, but may not be aware >of problems one can have doing GMS with periodic acid oxidation - it works >fine for basement membrane staining in 2 um kidney sections i.e. Jones >methenamine silver, but for accurate results, chromic acid may be the >better choice for fungus work. There was a huge discussion on Histonet on >this subject a few years back. > >Also, you still have to collect the silver solution as hazardous waste so >one more thing should not be too much of a problem. I do feel for those who >have to pay for this collection/waste disposal, we are fortunate our >university picks up that cost. > > > >At 03:03 PM 5/5/2004 -0500, you wrote: > > >>In an attempt to remove hazardous chromic acid from the lab, I vaguely >>remember replacing the Chromic Acid with 5% Periodic acid instead for the >> >> >GMS. > > >>Has anyone else done this? If my memory serves me correctly - it works >> >> >fine - > > >>but I am old . . . . er. (must be from all the teenagers at home). >> >>Jacqueline M. O'Connor HT(ASCP) >>Abbott Laboratories >>Global Pharmaceutical Research and Development >>Discovery Chemotherapeutics >> >> >> > > >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 kgrobert <@t> rci.rutgers.edu Thu May 6 08:56:52 2004 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Grocott's Methenamine Silver Stain In-Reply-To: <20040505204150.19445.qmail@web40908.mail.yahoo.com> References: <20040505204150.19445.qmail@web40908.mail.yahoo.com> Message-ID: <409A4424.5060803@rci.rutgers.edu> Mmmm, good point. I made up the borax fresh, but the source bottle I used is quite old, I think. Another addition to the "dump" list for haz waste pickup! Thanks! Kathleen Amy Porter wrote: > I would be suspicious about the borax. We do decalcified mouse > turbinates with this method for fungal sinusitis. Any time I have no > or weak reaction i make up fresh borax. Since you are making > everything up fresh, ?????. If your control hasn't been decaled then > it seems like that should be working if all your solutions are fresh. > I use a water-bath and prewarm my working silver before i add my > slides. Hope you figure out what is wrong, very frustrating. > > Kathleen Roberts wrote: > > Hey, all- > > I've been trying to stain some FFPE slides of decalcified rat sinuses > for fungi using the AFIP protocol for Grocott's stain, and despite > using > fresh reagents, the positive control and unknowns do not stain at > all, > except for some random deposition of silver that does not even > begin to > look like fungi. The positive control is loaded with Candida, and can > be seen with H&E easily. > > So far, we don't see any fungi in the rat sinuses and lungs (they > were > sneezing, and the bedding came back positive for Aspergillus) with > H&E > staining, which is why we decided to try the Grocott's-one way or > another we should be able to visualize it in the tissues. > > Any ideas why the Candida-loaded positive control would not stain > with > Grocott's? Right now I am thinking that perhaps they didn't sit long > enough in the 4% chromi c acid, even though the protocol says to > soak the > slides for an hour to oxidize them. Either that or a longer soak > in the > methenamine-silver solution at 60 degrees C (also for one hour). > > Thanks in advance for your help- > Kathleen Roberts > Principal Lab Technician > Neurotoxicology Labs > Dept of Pharmacology & Toxicology > Ernest Mario School of Pharmacy > Rutgers University > 41 B Gordon Rd > Piscataway, NJ 08854 > > > _______________________________________________ > 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!? > Win a $20,000 Career Makeover at Yahoo! HotJobs > From Terry.Marshall <@t> rothgen.nhs.uk Thu May 6 09:13:59 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Old farts reminiscing (was Polyester wax (with earlier citations)) Message-ID: About 8 years back we had a short discussion on whether sections are better now than in the old days. The overwhelming vote (from which I dissented) was yes. Thinner - yes. That's about the only improvement, and even that's not always a desirable quality. Anyway, I miss the older generation of histotechnicians (as we used to call them over here). My first encounter was with one Des Brady in Sheffield. He was great. If you asked for a special stain, he would say "do you want it positive or negative boss?" That's what I call skill:-) 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: Ian Montgomery [mailto:ian.montgomery@bio.gla.ac.uk] Sent: 06 May 2004 11:55 To: histonet@lists.utsouthwestern.edu Subject: Fwd: Re: [Histonet] Re: Polyester wax (with earlier citations) Here I am in sunny Glasgow living in a time warp. Automation, what's that? Hand process, make my own stains, sharpen knives, name it and I'm still doing it. Well apart from wooden blocks, gave that up 30 odd years ago. I'm such a sad case that if you ask me what a particular chemical in my store is for I'll tell you exactly and how much is used. Sad thing is, I'll be retiring shortly and all the knowledge will go with me. Ian. >Hi Paul, > >Lots of good times; >but you forgot about the blistered finger tips from sticking those wax >blocks on wooden blocks(the smaller the block-the bigger the blister!), and >all the hand processing, which mainly involved much frantic dashing about >twirling multiple pots of tissue to provide agitation! >Who can forget about mouth pipetting whilst eating sandwiches at the >staining bench and brewing the tea over a bunsen during waiting times! >If I recall correctly, the pipes were full of either St. Bruno flake or >'Herbal' tobacco and between chuffs of smoke, most of the conversation was >punctuated by phrases such as "eh up", "y'whot", "gerroff" and "owd ya >tight", interspersed with "cool man", "crazy daddy" and "all gone". Ahhh... >sweet memories. > >Bryan Hewlett >Hamilton, Ont. >Canada >(formerly Nottingham, England) > >----- Original Message ----- >From: "Paul Bradbury" >To: ; "HistoNet Server" > >Sent: Wednesday, May 05, 2004 3:32 PM >Subject: Re: [Histonet] Re: Polyester wax (with earlier citations) > > > > Hi Steve, > > > > Your descriptions bring back a flood of memories ... the wax tea pot, > > Leukhart's embedding rings, sticking the blocks onto wooden blocks, > > taking them off again at the end of the day, etc. Safety precautions had > > not even been invented in those days. When I first started in Histology, > > there were five of us in the lab and every single one smoked a pipe. So, > > embedding, trimming and sticking on the blocks involved three of us ... > > all chuffing out smoke. The conversations that took place during these > > times were priceless. Sadly, this opportunity was lost with the advent > > of automated embedding centres.. > > > > All solvents went down the drain, old specimens were dumped into the > > sink to allow the formalin to drain away. There was no fume hood, so the > > formalin fumes were thick enough to cut with a knife. In retrospect, > > dumping solvents and fixatives down the drain was not the best idea!... > > but at the time, that was standard practice. However, despite these > > horrendous practices, we are all still alive and well, and all went on > > to accept senior positions around the world.. > > > > There were no productivity units to count, no QC/QA demands (apart from > > self-imposed ones), no intrusions from mis-guided administrators. We had > > time to work on our own projects, investigate new procedures, and read > > journals looking for new methods. We made all of our own reagents from > > scratch (hematoxylin, Schiff's reagent, fixatives, etc) We sharpened our > > own knives. The camaraderie was wonderful, there was no bitching or > > whining, going for a beer at lunchtime was a routine practice. We did a > > great job, we went home happy, and provided great service > > > > I would not give up the new developments in Histology > > (immunohistochemistry, monoclonal antibodies, disposable knives, or > > automated stainers, etc ) they have produced quantum leaps in quality > > and diagnostic accuracy, but I sometimes I despair that the new > > generation of technologists have missed out on an invaluable learning > > experience. > > > > I firmly believe that I am a better histologist from my experience ... > > if something didn't work there was no service rep to call for advice, it > > was up to us to figure it out. The most respected "mentors" on the > > Histonet (who I won't name to avoid embarrasing them, but you know who I > > mean) are all the product of this bygone age of Histology. The Histonet > > serves a great purpose as a knowledge base and resource for advice, but > > there is no substitute for self-motivated learning. The books and > > journals are all out there ... waiting. > > > > Okay, I am done. Time to get down off my soapbox. The sermon is ended, > > > > Paul Bradbury > > Kamloops, BC > > Canada > > (formerly Nottingham, England) > > > > > > > > > > > > > > > > > > _______________________________________________ > > 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 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 JWEEMS <@t> sjha.org Thu May 6 09:26:19 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Old farts reminiscing (was Polyester wax (with ear lier citations)) Message-ID: But years ago we could actually process the tissue adequately - we didn't process tissue that wasn't fixed. And we had mercury in the hematoxylin which made it practically indestructable. And you could watch what you were staining since it was manual. I think I may have been able to make a positive special stain negative, but it would have been by accident - not by the skill level of your mentor! Ah the good ole days. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr, Consultant Histopathologist Sent: Thursday, May 06, 2004 10:14 AM To: Ian Montgomery; histonet@lists.utsouthwestern.edu Subject: [Histonet] Old farts reminiscing (was Polyester wax (with earlier citations)) About 8 years back we had a short discussion on whether sections are better now than in the old days. The overwhelming vote (from which I dissented) was yes. Thinner - yes. That's about the only improvement, and even that's not always a desirable quality. Anyway, I miss the older generation of histotechnicians (as we used to call them over here). My first encounter was with one Des Brady in Sheffield. He was great. If you asked for a special stain, he would say "do you want it positive or negative boss?" That's what I call skill:-) 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: Ian Montgomery [mailto:ian.montgomery@bio.gla.ac.uk] Sent: 06 May 2004 11:55 To: histonet@lists.utsouthwestern.edu Subject: Fwd: Re: [Histonet] Re: Polyester wax (with earlier citations) Here I am in sunny Glasgow living in a time warp. Automation, what's that? Hand process, make my own stains, sharpen knives, name it and I'm still doing it. Well apart from wooden blocks, gave that up 30 odd years ago. I'm such a sad case that if you ask me what a particular chemical in my store is for I'll tell you exactly and how much is used. Sad thing is, I'll be retiring shortly and all the knowledge will go with me. Ian. >Hi Paul, > >Lots of good times; >but you forgot about the blistered finger tips from sticking those wax >blocks on wooden blocks(the smaller the block-the bigger the blister!), and >all the hand processing, which mainly involved much frantic dashing about >twirling multiple pots of tissue to provide agitation! >Who can forget about mouth pipetting whilst eating sandwiches at the >staining bench and brewing the tea over a bunsen during waiting times! >If I recall correctly, the pipes were full of either St. Bruno flake or >'Herbal' tobacco and between chuffs of smoke, most of the conversation was >punctuated by phrases such as "eh up", "y'whot", "gerroff" and "owd ya >tight", interspersed with "cool man", "crazy daddy" and "all gone". Ahhh... >sweet memories. > >Bryan Hewlett >Hamilton, Ont. >Canada >(formerly Nottingham, England) > >----- Original Message ----- >From: "Paul Bradbury" >To: ; "HistoNet Server" > >Sent: Wednesday, May 05, 2004 3:32 PM >Subject: Re: [Histonet] Re: Polyester wax (with earlier citations) > > > > Hi Steve, > > > > Your descriptions bring back a flood of memories ... the wax tea pot, > > Leukhart's embedding rings, sticking the blocks onto wooden blocks, > > taking them off again at the end of the day, etc. Safety precautions had > > not even been invented in those days. When I first started in Histology, > > there were five of us in the lab and every single one smoked a pipe. So, > > embedding, trimming and sticking on the blocks involved three of us ... > > all chuffing out smoke. The conversations that took place during these > > times were priceless. Sadly, this opportunity was lost with the advent > > of automated embedding centres.. > > > > All solvents went down the drain, old specimens were dumped into the > > sink to allow the formalin to drain away. There was no fume hood, so the > > formalin fumes were thick enough to cut with a knife. In retrospect, > > dumping solvents and fixatives down the drain was not the best idea!... > > but at the time, that was standard practice. However, despite these > > horrendous practices, we are all still alive and well, and all went on > > to accept senior positions around the world.. > > > > There were no productivity units to count, no QC/QA demands (apart from > > self-imposed ones), no intrusions from mis-guided administrators. We had > > time to work on our own projects, investigate new procedures, and read > > journals looking for new methods. We made all of our own reagents from > > scratch (hematoxylin, Schiff's reagent, fixatives, etc) We sharpened our > > own knives. The camaraderie was wonderful, there was no bitching or > > whining, going for a beer at lunchtime was a routine practice. We did a > > great job, we went home happy, and provided great service > > > > I would not give up the new developments in Histology > > (immunohistochemistry, monoclonal antibodies, disposable knives, or > > automated stainers, etc ) they have produced quantum leaps in quality > > and diagnostic accuracy, but I sometimes I despair that the new > > generation of technologists have missed out on an invaluable learning > > experience. > > > > I firmly believe that I am a better histologist from my experience ... > > if something didn't work there was no service rep to call for advice, it > > was up to us to figure it out. The most respected "mentors" on the > > Histonet (who I won't name to avoid embarrasing them, but you know who I > > mean) are all the product of this bygone age of Histology. The Histonet > > serves a great purpose as a knowledge base and resource for advice, but > > there is no substitute for self-motivated learning. The books and > > journals are all out there ... waiting. > > > > Okay, I am done. Time to get down off my soapbox. The sermon is ended, > > > > Paul Bradbury > > Kamloops, BC > > Canada > > (formerly Nottingham, England) > > > > > > > > > > > > > > > > > > _______________________________________________ > > 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 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 _______________________________________________ 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 histology.bc <@t> shaw.ca Thu May 6 09:18:21 2004 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Re: Polyester wax (with earlier citations) In-Reply-To: References: Message-ID: <409A492D.3080308@shaw.ca> Hi Steve, I was the Chief Tech at Nottingham City Hospital, so I knew all the staff at Queen's Medical Centre. I know Neil Hand well, he started as a student at QMC while I was at the City. Alan Stevens, Janet Palmer, Keith Miller, the brothers Keith and Clive Gordon were all good friends of mine.Before the existing building was built, the Pathology Dept was housed in a row of ex-army huts. This was a very humble beginning for what is now a very well respected department. John Bancroft was the Chief Tech back in those days. We used to teach nightschool together in the labs at QMC or the City. This was always followed by several pints at the hospital pub ... that's another feature of hospital life that sadly seems to have gone. Darts tourmaments between the two labs were a regular event. Thinking about old pathologists reminded me of one at the City. The first automated knife sharpener we got was the original Shandon Elliot, this was the highlight of the department. This old pathologist was so excited about this new equipment that in his rush to see it he backed into two cars parked outside the lab, and fell over the curb. He then proceeded to sit on a stool and watch the plate rotate and the knife flip from side to side for the next four hours. Unsophisticated times. Food was big thing in our lab too. One of the staff always went to get tea and bacon sandwiches at 10 o'clock, these were then eaten while cutting sections. Getting bacon grease on the slides was sure way to make sure the sections floated off later. A dress code was in effect in our lab too. Shirt and tie was the rule of the day. Suits were the standard, apart from Saturdays when we wore sports jackets (more casual). We worked alternate Saturdays, just until lunch time, to get any urgent specimens done. One very quiet morning, we made a cannon from a length of 1" glass tube, a length of hose and the CO2 tank from the freezing microtome, bals of wax were the ammunition. Boy, did that thing work well! They were the days ... Cheers, Paul From gcallis <@t> montana.edu Thu May 6 09:48:49 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] The unjoys of working with cryo rat turbinates In-Reply-To: <20040505230634.49439.qmail@web13310.mail.yahoo.com> Message-ID: <3.0.6.32.20040506084849.00c058a8@gemini.msu.montana.edu> Gareth, This is the reason we invested in Cryojane for the majority of undecalcified or decalcified FS and difficult soft tissue FS work. If you want to do antigen retrieval on any kind of frozen section of bone, calcified or decalcified or even soft tissues, they will love to fall off the plus charge slides and annoy you into tears or fits of bad language. I do have one fellow who decalcifies in EDTA on periodate/lysine/paraformaldehyde (PLP) perfused, then immersion fixed mouse turbinates at 4C, with one change of this fixative. He cryoprotects during decalcification with 10 % sucrose/1.25% EDTA, tetrasodium in PBS until head is done (1 to 2 weeks, rat will take longer than mouse) snap freezes and mounts his FS on plus charge slides. We can H&E on these but the rinsing is minimal/gentle not with running water. We also use Richard Allan progressive Hematoxylin 1, do not clarify, blue, rinse briefly in tap water, go to 70%, eosin, etc, etc. However, for IHC, he NEVER does retrieval but if you antigen is compromised by cross linking, you may be better off with fixation then EDTA decalcification - NO frozen sections at all IF you need to do retrieval. Processing rat turbinates needs to be done on an extended processing schedule - if you need one, let me know, it is easy to set up. Discouraging but possible, tests your patience,creativity, maybe the pocketbook if you buy Cryojane. I'm off to undecalcified mouse snout FS this A.M. but Cryojane is assisting 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 Thu May 6 10:03:04 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] cryoprotection before MMA embedding at =?iso-8859-1?Q?-20=B0C?= In-Reply-To: <71FE4E26.5FC3474F.0005167B@aol.com> Message-ID: <3.0.6.32.20040506090304.00c16ef8@gemini.msu.montana.edu> I have never cryoprotected tissues embedded in (Methylmethacrylate) MMA nor seen a protocol calling for this step with MMA. If you have a publication reference on this, it would be nice to see their rationale for doing this. Sucrose cryoprotection is used to reduce water ice crystal formation in fixed tissue destined for cryosectioning, aka frozen sections. In general, after tissue/cells are fixed, you should start dehydration - I am trying to fathom why sucrose would be used with MMA protocol, awaiting more comments on this. The reason you put MMA in the cold is to prevent it from polymerizing suddenly as you go through the final steps infiltration steps. On occasion, it can suddenly turn into solid plastic - something you do not want to happen. Cold temperatures during dehydration and clearing may help preserve antigens IF you need to IHC, but you still need to remove MMA before this kind of staining. Most people dehydrate, clear at room temperature, but infiltrate at 4C with MMA to keep the plastic mixture from polymerizing before tissues/cells are completely infiltrated with unpolymerized MMA. Because heat is given off during polymerization of MMA, cold embedding is used to protect antigens on cells IF immunostaining is a goal and can be done at 4C, refrigerator temperatures also. You may have lost the tissue somewhere in the changes of reagents. At 05:59 AM 5/6/2004 -0400, you wrote: >Hi everyone >i use a protocol for MMA embedding at 20?C since a few months. > and this protocol says to put simples in phosphate buffer 7.4 with 10% sucrose overnight at 4?C after fixation in paraformald?hyde 4% for 3 days and before dehydratation and preimpregnation at 4?C, emebdding is done at -20?C. >Last time i forget the step of cryoprotection with sucrose, and after cutting and staining my embedding simples, i didn't see any cells, no one cell !!! i did never see that before !!!! >Do you think that is because of no cryoprotection ? >Does a lack cryoprotection induce disparition of cells ? the tissue was very thin... >any help would be very usefull >Thank you very much >Myriam Baali >Natural implant > >_______________________________________________ >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 ttroyer <@t> pcllab.com Thu May 6 10:10:46 2004 From: ttroyer <@t> pcllab.com (Travis Troyer) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] G raded Alcohols Message-ID: <008301c4337c$4f2e9270$6601010a@Peterson.local> I have a question regarding the graded alcohols used in both the processor and autostainer. We are trying out a recycler and the results we get back are around 92%. I was curious how this would effect the result going from 92% to 100% on the processor. We run all of our tissue together through the processor so we have biopsies, breast cases, and uterus blocks all in the same run. Thank you for your assistance, Travis Troyer Peterson Clinical Lab From vandries <@t> vub.ac.be Thu May 6 10:42:23 2004 From: vandries <@t> vub.ac.be (Veronique Andriessen) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] double immuno staining In-Reply-To: Message-ID: Thanks everyone for all the kind replies! I am definately gonna try the PLP fixation. I would love to use acetone fixation, but this is not possible for the somatostatin receptor. It will destroy most of the cell membranes. As for antigen retrieval, I've tried HIER at pH 6 and 9 and EIER with pronase and trypsin at different time intervals, dilutions and temperatures. 10 minutes at RT with trypsin-EDTA gives the best result (morphology vs staining intensity) for this fixation. I am also going to look for another desmin antibody clone to mix with the one I'm already using. Hopefuly this will enhance the signal. Sincerely yours, Veronique Andriessen BAS Lab. Molecular Liver Cell Biology, Free University Brussels (VUB) Belgium -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Veronique Andriessen Sent: woensdag 5 mei 2004 16:19 To: Histonet Subject: [Histonet] double immuno staining Hi folks, I need some help. I am struggling with a double immunostaining for somatostatin receptor and desmin. All my previous double stainings worked beautifully, but they were performed on fresh frozen acetone fixed pancreatic tissue. For staining of somatostatin receptor I have to fix with formaldehyde. I use 20 minutes 0.75% PFA perfused tissue which is snap frozen embedded in tissue freezing medium and cut on a cryostat. I was advised to do it this way to prevent loss of antigenicity. Morphology is quite good. This fixation works for the somatostatin receptor staining, but it kills the desmin signal. I was able to retrieve most of the desmin signal by EIER for 10 minutes with trypsin-EDTA, but now the morphology is awful. Can someone give me some advice? Sincerely yours, Veronique Andriessen BAS Lab. Molecular Liver Cell Biology, Free University Brussels (VUB)Belgium _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lizchlipala <@t> premierhistology.com Thu May 6 10:45:00 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] SMI 31 and SMI 32 Immunohistochemistry for Neurofiliaments Message-ID: <000001c43381$1a9a63e0$74d48a80@LIZ> Hello Everyone I was hoping someone might be able to help me out on this. I have a client who wants me to perform SMI 31 and SMI 32 on rat spinal cord samples. I know that these antibodies are from Sternberger/Meyer Immunochemicals. I have searched on the internet for a web page and in MSN yellow pages to try to find some contact information about this company but have come up with nothing. Does anyone know their web page address or phone number? Also while I'm asking the few references I have looked at use these antibodies on fixed vibratome or cryostat sections (free floating immunos). Can these be performed on FFPE sections and also are there other antibodies available that will stain the same thing. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP) 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 MCBD, Room A3B40 Boulder, Colorado 80309 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From chris.goodall <@t> bristol.ac.uk Thu May 6 11:09:21 2004 From: chris.goodall <@t> bristol.ac.uk (anclg) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] the good old days Message-ID: <000301c43384$7e43aac0$8f6ede89@hist143> Hi everyone, you made me laugh........I`m in a time warp here too in not so sunny Bristol. I do have a dip and dunk processor,circa 1980, but can well remember hand processing, when I first started in the lab. in 1970, we were using benzene as the solvent and I used to lift the lid of the wax oven where the pots of benzene / wax and wax pots were kept and think what a lovely smell it was! I still sharpen my knives on an old Shandon Elliott,including the D profile tungsten carbide knives. I can certainly remember that awful sinking feeling as I waited, breath held ,watching the knife flip over only to see the thin slivers of copper shaved from the plate surface again.It`s really only in the last couple of years that I`ve used tissue cassettes for everything and even now use Leukhart moulds and wooden blocks for large bone blocks, and all my wax is still warmed in a jug over a bunsen burner. I make all my own stains and fixatives, all prepared under a fume hood now though and with all the proper safety precautions. I have advanced now into the age of immunohistochemistry,in situ hybridisation, plastic embedding etc, but some old habits die hard and will probably remain with me until I retire. Of course, if anyone would like to donate a tissue processor, embedding station, automated stainer ........sigh...... Chris Goodall From Terry.Marshall <@t> rothgen.nhs.uk Thu May 6 10:53:54 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Re: Polyester wax (with earlier citations) Message-ID: Paul cites: "One very quiet morning, we made a cannon from a length of 1" glass tube, a length of hose and the CO2 tank from the freezing microtome, bals of wax were the ammunition. Boy, did that thing work well!" In Stoke (Kemlo will remember well) the perpetual sport in the summer was shooting wasps in the lab with rubber bands. Poor wasps. Can't remember what the winter sport was - just waiting for the wasps I guess. 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: Paul Bradbury [mailto:histology.bc@shaw.ca] Sent: 06 May 2004 15:18 To: Stephen.Eyres@sanofi-synthelabo.com; HistoNet Server Subject: Re: [Histonet] Re: Polyester wax (with earlier citations) Hi Steve, I was the Chief Tech at Nottingham City Hospital, so I knew all the staff at Queen's Medical Centre. I know Neil Hand well, he started as a student at QMC while I was at the City. Alan Stevens, Janet Palmer, Keith Miller, the brothers Keith and Clive Gordon were all good friends of mine.Before the existing building was built, the Pathology Dept was housed in a row of ex-army huts. This was a very humble beginning for what is now a very well respected department. John Bancroft was the Chief Tech back in those days. We used to teach nightschool together in the labs at QMC or the City. This was always followed by several pints at the hospital pub ... that's another feature of hospital life that sadly seems to have gone. Darts tourmaments between the two labs were a regular event. Thinking about old pathologists reminded me of one at the City. The first automated knife sharpener we got was the original Shandon Elliot, this was the highlight of the department. This old pathologist was so excited about this new equipment that in his rush to see it he backed into two cars parked outside the lab, and fell over the curb. He then proceeded to sit on a stool and watch the plate rotate and the knife flip from side to side for the next four hours. Unsophisticated times. Food was big thing in our lab too. One of the staff always went to get tea and bacon sandwiches at 10 o'clock, these were then eaten while cutting sections. Getting bacon grease on the slides was sure way to make sure the sections floated off later. A dress code was in effect in our lab too. Shirt and tie was the rule of the day. Suits were the standard, apart from Saturdays when we wore sports jackets (more casual). We worked alternate Saturdays, just until lunch time, to get any urgent specimens done. One very quiet morning, we made a cannon from a length of 1" glass tube, a length of hose and the CO2 tank from the freezing microtome, bals of wax were the ammunition. Boy, did that thing work well! They were the days ... Cheers, Paul _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Stephen.Eyres <@t> sanofi-synthelabo.com Thu May 6 10:46:48 2004 From: Stephen.Eyres <@t> sanofi-synthelabo.com (Stephen.Eyres@sanofi-synthelabo.com) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Re: Polyester wax (with earlier citations) Message-ID: Hi Paul, I've been to a couple of the pub sessions during the histology course. As I remember, John used to get bored and then retire to the pub to continue the discussion. We were lucky in that we only worked 1 in 3 Saturdays. Quite a few lads would go out for a session on friday straight from work. many a time we'd miss the last train, go and sleep in the path lab tearoom, to be awakened by the cleaners. The off to the shop for a sandwich and then back to work for a few hours.....Then down the pub for a few before drifting off home. Did you do a block release HNC? I to Bristol on the 3x 6 week blocks. That was a fun time. I remeber starting in the labs in September, leaving for an op on my hand in 2 weeks later - not work related- returning in October and then going on block at the beginning of january having only done 1 manual H&E. What a mess I made at first doing special stains during the 3 hour practicals. I'd be putting schiff's on an MSB slide, Phosphotungstic on a retic. Mind, it is worth pointing out to anyone reading these that although things were very different 20-30 years ago in the histo lab, we were very busy and would work like stink in the mornings to get things signed out so we could do specials and 'play' with new stains, or in my case try to convince someone they ought to try cutting polyester wax blocks!! I never did succeed. That particular 'right of passage' was abandoned. Cheers Steve Paul Bradbury To: Stephen Eyres/GB-ALNWICK/RESEARCH/SANOFI@Research Sent by: HistoNet Server histonet-bounces@lists.utsouth cc: western.edu Subject: Re: [Histonet] Re: Polyester wax (with earlier citations) 06/05/2004 15:18 Hi Steve, I was the Chief Tech at Nottingham City Hospital, so I knew all the staff at Queen's Medical Centre. I know Neil Hand well, he started as a student at QMC while I was at the City. Alan Stevens, Janet Palmer, Keith Miller, the brothers Keith and Clive Gordon were all good friends of mine.Before the existing building was built, the Pathology Dept was housed in a row of ex-army huts. This was a very humble beginning for what is now a very well respected department. John Bancroft was the Chief Tech back in those days. We used to teach nightschool together in the labs at QMC or the City. This was always followed by several pints at the hospital pub ... that's another feature of hospital life that sadly seems to have gone. Darts tourmaments between the two labs were a regular event. Thinking about old pathologists reminded me of one at the City. The first automated knife sharpener we got was the original Shandon Elliot, this was the highlight of the department. This old pathologist was so excited about this new equipment that in his rush to see it he backed into two cars parked outside the lab, and fell over the curb. He then proceeded to sit on a stool and watch the plate rotate and the knife flip from side to side for the next four hours. Unsophisticated times. Food was big thing in our lab too. One of the staff always went to get tea and bacon sandwiches at 10 o'clock, these were then eaten while cutting sections. Getting bacon grease on the slides was sure way to make sure the sections floated off later. A dress code was in effect in our lab too. Shirt and tie was the rule of the day. Suits were the standard, apart from Saturdays when we wore sports jackets (more casual). We worked alternate Saturdays, just until lunch time, to get any urgent specimens done. One very quiet morning, we made a cannon from a length of 1" glass tube, a length of hose and the CO2 tank from the freezing microtome, bals of wax were the ammunition. Boy, did that thing work well! They were the days ... Cheers, Paul _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo <@t> tiscali.co.uk Thu May 6 11:11:50 2004 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Old farts reminiscing (was Polyester wax (with earliercitations)) In-Reply-To: Message-ID: <001001c43384$dafdc3e0$f8cce150@KEMLOS> Anyway, I miss the older generation of histotechnicians (as we used to call them over here). My first encounter was with one Des Brady in Sheffield. He was great. If you asked for a special stain, he would say "do you want it positive or negative boss?" That's what I call skill:-) Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. From asmith <@t> mail.barry.edu Thu May 6 11:39:26 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] lymph nodes Message-ID: <494304423C63E246A5CF87A3AEEB577011B5E9@bumail01.barrynet.barry.edu> Most drugstores still carry old fashioned single edge razor blades which have a metal strip folded over the dull edge. They are much easier to dissect with than the disposable microtome blades. I use them for block trimming. 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 Marshall Terry Dr,Consultant Histopathologist Sent: Wednesday, May 05, 2004 11:37 AM To: Mike Bromley; Histonet (E-mail) Subject: RE: [Histonet] lymph nodes I find for this sort of thing, disposable microtome blades are ideal. Holders are available, but I neither use nor recommend them. 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: Mike Bromley [mailto:M.Bromley@dgri.scot.nhs.uk] Sent: 05 May 2004 16:26 To: Histonet (E-mail) Subject: [Histonet] lymph nodes Hi Everyone I am after a very thin blade to slice unfixed lymph nodes, does anyone have any ideas? Ideally some kind of holder for old style razor blades would be the answer. Best Wishes Mike Bromley Chief Biomedical Scientist Pathology Dumfries & Galloway Royal Infirmary Scotland, 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 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 Terry.Marshall <@t> rothgen.nhs.uk Thu May 6 11:55:04 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] lymph nodes Message-ID: Not so. Unless you have superior blades in the US, they are considerably more blunt than a Feather blade. Furthermore, their short length precludes a cut, i.e. a longitudinal stroke with the blade. They will be fine for block trimming. The purpose about which we are discussing, is the slicing of fresh nodes, an objective only achieved by a proper cutting stroke with the sharpest available blade. 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: Smith, Allen [mailto:asmith@mail.barry.edu] Sent: 06 May 2004 17:39 To: Marshall Terry Dr, Consultant Histopathologist Cc: histonet@pathology.swmed.edu Subject: RE: [Histonet] lymph nodes Most drugstores still carry old fashioned single edge razor blades which have a metal strip folded over the dull edge. They are much easier to dissect with than the disposable microtome blades. I use them for block trimming. 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 Marshall Terry Dr,Consultant Histopathologist Sent: Wednesday, May 05, 2004 11:37 AM To: Mike Bromley; Histonet (E-mail) Subject: RE: [Histonet] lymph nodes I find for this sort of thing, disposable microtome blades are ideal. Holders are available, but I neither use nor recommend them. 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: Mike Bromley [mailto:M.Bromley@dgri.scot.nhs.uk] Sent: 05 May 2004 16:26 To: Histonet (E-mail) Subject: [Histonet] lymph nodes Hi Everyone I am after a very thin blade to slice unfixed lymph nodes, does anyone have any ideas? Ideally some kind of holder for old style razor blades would be the answer. Best Wishes Mike Bromley Chief Biomedical Scientist Pathology Dumfries & Galloway Royal Infirmary Scotland, 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 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 laurie.colbert <@t> huntingtonhospital.com Thu May 6 12:10:45 2004 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] lymph nodes Message-ID: <0BE6ADFAE4E7E04496BF21ABD34662801D0CD3@EXCHANGE1.huntingtonhospital.com> We use a double-edged razor blade. It is longer than a normal razor blade and it comes with a handle. We order through Allegiance. The catalog # for the handle is D2877 and the # for the blades (100/bx) is D2878-2C. Laurie Colbert -----Original Message----- From: Smith, Allen [mailto:asmith@mail.barry.edu] Sent: Thursday, May 06, 2004 9:39 AM To: Marshall Terry Dr,Consultant Histopathologist Cc: histonet@pathology.swmed.edu Subject: RE: [Histonet] lymph nodes Most drugstores still carry old fashioned single edge razor blades which have a metal strip folded over the dull edge. They are much easier to dissect with than the disposable microtome blades. I use them for block trimming. 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 Marshall Terry Dr,Consultant Histopathologist Sent: Wednesday, May 05, 2004 11:37 AM To: Mike Bromley; Histonet (E-mail) Subject: RE: [Histonet] lymph nodes I find for this sort of thing, disposable microtome blades are ideal. Holders are available, but I neither use nor recommend them. 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: Mike Bromley [mailto:M.Bromley@dgri.scot.nhs.uk] Sent: 05 May 2004 16:26 To: Histonet (E-mail) Subject: [Histonet] lymph nodes Hi Everyone I am after a very thin blade to slice unfixed lymph nodes, does anyone have any ideas? Ideally some kind of holder for old style razor blades would be the answer. Best Wishes Mike Bromley Chief Biomedical Scientist Pathology Dumfries & Galloway Royal Infirmary Scotland, 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 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) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Myri37 <@t> aol.com Thu May 6 12:30:05 2004 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:22:55 2005 Subject: =?ISO-8859-1?Q?Re:=20[Histonet]=20cryoprotection=20before=20MMA=20?= =?ISO-8859-1?Q?embedding=20at=20=20-20=B0C?= Message-ID: <1e8.1fb28df8.2dcbd01d@aol.com> I found this protocol in reference of a publication of Erben RG, 1997 it's about "embedding of bone in methylmetacrylate an improved method suitable for bone in histomorphometry, histochemitry, and immunohistochemitry" embedding is done at -20?C that is why i think it needs a cryoprotection. it's the first time a lost all tissue, and i dont understand why. thank you Myriam From gcallis <@t> montana.edu Thu May 6 12:35:11 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] SMI 31 and SMI 32 Immunohistochemistry forNeurofiliaments In-Reply-To: <000001c43381$1a9a63e0$74d48a80@LIZ> Message-ID: <3.0.6.32.20040506113511.00c16ef8@gemini.msu.montana.edu> http://home.att.net/~sternbmonoc/home.htm Or just do a search for Sternberger Monoclonals Incorporated, website come up immediately. 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 Thu May 6 12:44:38 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] cryoprotection before MMA embedding at =?iso-8859-1?Q?=2D20=B0C?= References: <1e8.1fb28df8.2dcbd01d@aol.com> Message-ID: <409A7986.B752DD54@uwo.ca> If the -20C is for the methyl methacrylate infiltration you won't need cryoprotection because by that stage there shouldn't be any water in the specimens. You have to go down below -114C to freeze absolute alcohol or -94C to freeze acetone. -- ------------------------------- 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 _______________________________ Myri37@aol.com wrote: > > I found this protocol in reference of a publication of Erben RG, 1997 > it's about "embedding of bone in methylmetacrylate an improved method > suitable for bone in histomorphometry, histochemitry, and immunohistochemitry" > embedding is done at -20?C that is why i think it needs a cryoprotection. > it's the first time a lost all tissue, and i dont understand why. > thank you > Myriam > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lizchlipala <@t> premierhistology.com Thu May 6 12:50:23 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Thanks for Info on SMI 31 and SMI 32 Message-ID: <000e01c43392$9dfd9520$74d48a80@LIZ> Thanks for all the responses I got what I needed. Liz Elizabeth A. Chlipala, BS, HTL(ASCP) 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 MCBD, Room A3B40 Boulder, Colorado 80309 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From Sandra.Etheridge <@t> gems8.gov.bc.ca Thu May 6 13:05:10 2004 From: Sandra.Etheridge <@t> gems8.gov.bc.ca (Etheridge, Sandra AGF:EX) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Wright-Giemsa stain for smears Message-ID: <424FAC08A8CDFE4BADD6497AE75B5E2F0E0A8684@atlas.gov.bc.ca> Hello fellow Histonetters! I am looking for a manual staining procedure for cytology or blood smears using the commercial Wright-Giemsa stain. I was also wondering if it can be used on regular tissue sections. I believe the method is quite fast and our current method does not give adaquate nuclear staining. Our reagent expired in December '03, so that could also be the problem. Your feedback would be appreciated. Sandra Etheridge Animal Health Center BC Ministry of Agriculture, Food, and Fisheries Abbotsford, BC From JPCOLEMA <@t> sentara.com Thu May 6 13:08:29 2004 From: JPCOLEMA <@t> sentara.com (JOHN COLEMAN) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Chromic acid vs periodic acid, old times, desmin IHC, Message-ID: For GMS in our lab we tried both Chromic and Periodic and found that because of the weakness of the Periodic we got positive non specific fiber and histiocyte staining in our Pneumocystis GMS slides that our pathologists didn't like to see. Stronger oxidation= cleaner stain (for us) and better contrast of target items. How about doing animal surgery without gloves, having a charcoal grill running burgers inside the morgue room on the spare table under the vent, "lunch and learn" brain cuttings in the autopsy suite, practical jokes which included exploding purple iodine clouds from under the toilet seat, and body bags full of striped bass in the cold storage unit! Work uniforms? How about shorts and a cutoff t- shirt underneath a lab coat. (I think sometimes somebody skipped the pants altogether!) Desmin- for desmin IHC i've found a 95C bath of high PH + EDTA retrieval solution (Trilogy from Cellmarque) for 15 minutes and no trypsin= less morphologic ruination. (Trypsin + heat retrieval= mush nuclei and swollen collagen. ) Desmin from Biogenex, 1:80 dilution 30 min incubation. From JPCOLEMA <@t> sentara.com Thu May 6 13:12:45 2004 From: JPCOLEMA <@t> sentara.com (JOHN COLEMAN) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Graded alcohols Message-ID: Going from 92- 100. Option 1)- 3 stations of 100 to compensate for the extra water. Option 2, mix equal amounts of 92 and 100 to get 96 and use this between 92 and 100. (if you have 100, use it to Boost the proof of your 92 ) JPJC From pam <@t> ategra.com Thu May 6 13:36:02 2004 From: pam <@t> ategra.com (Pam Barker (extension 234)) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Supervisory opportunities in Histology Message-ID: Histonetters - Are you a Histology Supervisor Seeking a Day Shift Position Mon-Fri Only Position ? I am presently on a search for several of my best clients nationwide who are seeking to hire a Histology Supervisors. These are MON-FRI ONLY - D A Y S ONLY. (no working on weekends or holidays ! ) I have positions in AL, CA, MD, FL, MO and CO. Supervisor Positions are for fulltime permanent employees. They pay excellent employee benefits (paid vacation, sick time, holiday pay, health insurance.) If you are interested, please first send me your resume first, then call me at 800-466-9919 x234. I will keep your resume confidential and will not release it without your permission. My services are at no charge to you. 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 From Solverboy <@t> aol.com Thu May 6 14:35:38 2004 From: Solverboy <@t> aol.com (Solverboy@aol.com) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Cracking tissue Message-ID: <1EA9184F.5271BD8D.0293A525@aol.com> Hello All, I am cutting transverse sections of fixed, frozen rat heart on a cryotstat. When I mount my sections immediately after cutting, the histology looks perfect, but within two minutes of warming(room temp.) onto Fisherbrand Superfrost Plus slides, the tissue begins to crack (more specifically the muscle fibers tend to seperate from one another). Does anyone know a solution to or a cause of this problem? Thanks in advance, Frederic Grau Dept. Neurobiology & Behavior S.U.N.Y. Stony Brook From Jackie.O'Connor <@t> abbott.com Thu May 6 14:35:56 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] old times Message-ID: OK - ! Let's go back into time about 35 years - - How about everyone smoking cigarettes and drinking coffee while cutting blocks - parties in the lab that included food, ethanol and orange juice (the pathologists's idea) - and a diener who could routinely be found sleeping in the walk-in morgue cooler? We had something on our processor between the acetone and xylene that was called "solvent 127". In 30 years, I've never been able to find out what that was. . .. maybe Soylent Green? We occasionally used toluene and dioxane to 'rush process' tissues (no ventillation). How about making bouin's solution with dry picric acid from a 5 pound jar. Sometimes if you spilled a little of the picric acid, and it was still there the next day, when you walked on it - it made little crackling noises. "JOHN COLEMAN" Sent by: histonet-bounces@lists.utsouthwestern.edu 05/06/2004 01:08 PM To: cc: Subject: [Histonet] Chromic acid vs periodic acid, old times, desmin IHC, For GMS in our lab we tried both Chromic and Periodic and found that because of the weakness of the Periodic we got positive non specific fiber and histiocyte staining in our Pneumocystis GMS slides that our pathologists didn't like to see. Stronger oxidation= cleaner stain (for us) and better contrast of target items. How about doing animal surgery without gloves, having a charcoal grill running burgers inside the morgue room on the spare table under the vent, "lunch and learn" brain cuttings in the autopsy suite, practical jokes which included exploding purple iodine clouds from under the toilet seat, and body bags full of striped bass in the cold storage unit! Work uniforms? How about shorts and a cutoff t- shirt underneath a lab coat. (I think sometimes somebody skipped the pants altogether!) Desmin- for desmin IHC i've found a 95C bath of high PH + EDTA retrieval solution (Trilogy from Cellmarque) for 15 minutes and no trypsin= less morphologic ruination. (Trypsin + heat retrieval= mush nuclei and swollen collagen. ) Desmin from Biogenex, 1:80 dilution 30 min incubation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lu_ze <@t> sbcglobal.net Thu May 6 15:53:54 2004 From: lu_ze <@t> sbcglobal.net (Ze Lu) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] AO 820 microtome questions Message-ID: <000b01c433ac$3f0944a0$0f1515ce@HOME2> Dear Histonet Friends, We just bought a used AO 820 microtome and wondered if anyone out there has an "Instruction Manual 820". If so, would it be possible to email it in pdf file? Also for this microtome, it came with a disposable blade holder. It seemed that the sectioning angle is fixed. Anybody knew how to change the angle? Thanks! Ze Lu, Ph. D. The Optimum Therapeutics, LLC From BlazekL <@t> childrensdayton.org Thu May 6 15:10:10 2004 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] old times Message-ID: Oh lord am I old. From JWEEMS <@t> sjha.org Thu May 6 15:27:54 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] old times Message-ID: but we're like fine wine....:>) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Linda Blazek Sent: Thursday, May 06, 2004 4:10 PM To: Jackie.O'Connor@abbott.com; JPCOLEMA@sentara.com Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] old times Oh lord am I old. _______________________________________________ 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 Joseph?s Health System, Inc. From Don.Birgerson <@t> leica-microsystems.com Thu May 6 15:30:01 2004 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] AO 820 microtome questions Message-ID: Dear Dr. Ze Lu, The American Optical Co. that first made the "820" Microtome in 1947, is now part of Leica. I have to guess you have a 820 that has a hinged lid and the original integrated disposable blade holder. If this is true, let me know and I can E mail a copy of the manual. Except for the locking lever kit, I am afraid parts are no longer available. Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Ze Lu" Sent by: To: histonet-bounces@lists.utsouth cc: western.edu Subject: [Histonet] AO 820 microtome questions 05/06/2004 03:53 PM Dear Histonet Friends, We just bought a used AO 820 microtome and wondered if anyone out there has an "Instruction Manual 820". If so, would it be possible to email it in pdf file? Also for this microtome, it came with a disposable blade holder. It seemed that the sectioning angle is fixed. Anybody knew how to change the angle? Thanks! Ze Lu, Ph. D. The Optimum Therapeutics, LLC _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Thu May 6 15:27:00 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] old times Message-ID: As long as you keep your cork - then even fine wine can turn into vinegar! :>) "Weems, Joyce" 05/06/2004 03:27 PM To: "'Linda Blazek'" , Jackie.O'Connor@abbott.com, JPCOLEMA@sentara.com cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] old times but we're like fine wine....:>) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Linda Blazek Sent: Thursday, May 06, 2004 4:10 PM To: Jackie.O'Connor@abbott.com; JPCOLEMA@sentara.com Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] old times Oh lord am I old. _______________________________________________ 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 Joseph's Health System, Inc. From jkiernan <@t> uwo.ca Thu May 6 15:42:15 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Wright-Giemsa stain for sections References: <424FAC08A8CDFE4BADD6497AE75B5E2F0E0A8684@atlas.gov.bc.ca> Message-ID: <409AA327.9283ADB5@uwo.ca> Wright's and Giemsa's are different stains, both used principally for blood and marrow to identify the various cell types. They are mixtures of blue cationic dyes (notably azure B) with eosin Y, an anionic dye. Both can be used on smears or sections. Giemsa gives excellent results with ordinary paraffin sections once you have found the ideal pH for the staining solution. Fixation affects the color balance, so you have to experiment the first time you do the method. After adequate formaldehyde fixation the best pH is often in the range 4.0 to 4.5. Solutions: 1. Acetate buffers, 0.2M from pH 3.5 to 7. (Only one of these will be needed when the optimum pH is known.) 2. Buffered water, made by 10-fold dilution of of the 0.2M buffer. The pH needs adjustment with drops of 5% acetic acid after dilution. 3. Giemsa stock solution. Usually this is bought ready-made. It should contain dyes certified by the Biological Stain Commission. (Ask the supplier.) Method. 1. Bring paraffin sections to water. 2. Put slides in acetate buffer for 5 minutes. 3. Pour off the buffer. (It can be reused.) 3. Add 1 ml Giemsa stock solution to 50 ml of buffered water in a beaker. Stir, then pour the diluted stain into the staining jar containing the slides. 4. Staining time is about 2 hours at 20C or about 15 minutes at 50-60C (microwave oven). 5. Rinse the slides in buffered water, blot dry and check with a microscope. If blue predominates, differentiate carefully in 0.01% acetic acid, rinse in buffered water and blot dry again. Too much blue means the pH of the buffer was too high. If red predominates, use a higher pH next time. 6. Either thoroughly air-dry the slides or dehydrate the almost dry sections in two changes, each 3 minutes, of n-butanol. 7. Clear in xylene and coverslip. In a correctly stained preparation nuclei and cytoplasmic RNA are blue to purple. Erythrocytes, collagen and keratin are pink. The different types of leukocyte are recognizable by their nuclei and cytoplasmic details. Bacteria are dark blue to purple. The preparations are more informative than sections stained with hemalum and eosin because more cytoplasmic detail is visible in a wider range of colors. The late R. D. Lillie perfected methods of this type as routine staining techniques for on a large scale (mechanized) in histopathology (see Lillie & Fullmer, 1976). Reference. Lillie RD, Fullmer H (1976) Histopathologic Technic and Practical Histochemistry. 4th ed. New York: McGraw-Hill, pp. 193-197. -- ------------------------------- 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 _______________________________ "Etheridge, Sandra AGF:EX" wrote: > > Hello fellow Histonetters! > > I am looking for a manual staining procedure for cytology or blood smears > using the commercial Wright-Giemsa stain. I was also wondering if it can be > used on regular tissue sections. I believe the method is quite fast and our > current method does not give adaquate nuclear staining. Our reagent expired > in December '03, so that could also be the problem. > > Your feedback would be appreciated. > > Sandra Etheridge > Animal Health Center > BC Ministry of Agriculture, Food, and Fisheries > Abbotsford, BC > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu May 6 15:59:31 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Cracking tissue In-Reply-To: <1EA9184F.5271BD8D.0293A525@aol.com> Message-ID: <3.0.6.32.20040506145931.00c06190@gemini.msu.montana.edu> Question: Are your prefixed hearts cryoprotected with 20 to 30% sucrose before snap freezing? At 03:35 PM 5/6/2004 -0400, you wrote: >Hello All, > I am cutting transverse sections of fixed, frozen rat heart on a cryotstat. When I mount my sections immediately after cutting, the histology looks perfect, but within two minutes of warming(room temp.) onto Fisherbrand Superfrost Plus slides, the tissue begins to crack (more specifically the muscle fibers tend to seperate from one another). Does anyone know a solution to or a cause of this problem? > >Thanks in advance, > >Frederic Grau >Dept. Neurobiology & Behavior >S.U.N.Y. Stony Brook > >_______________________________________________ >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 Thu May 6 16:20:19 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] AO 820 microtome questions In-Reply-To: <000b01c433ac$3f0944a0$0f1515ce@HOME2> Message-ID: <3.0.6.32.20040506152019.00c06190@gemini.msu.montana.edu> This microtome is such an old model, I don't think you will be able to get a pdf file manual. Leica Microsystems might have one sitting around in little booklet form. The disposable blade holder is for high profile blades only, and you cannot adjust the angle, however this preset angle was always good when we used it here, you don't need to fiddle with changing, it will work. At 03:53 PM 5/6/2004 -0500, you wrote: >Dear Histonet Friends, > >We just bought a used AO 820 microtome and wondered if anyone out there has >an "Instruction Manual 820". If so, would it be possible to email it in pdf file? Also for this microtome, it came with a disposable blade holder. It seemed that the sectioning angle is fixed. Anybody knew how to change the angle? >Thanks! > >Ze Lu, Ph. D. >The Optimum Therapeutics, LLC > > > > > >_______________________________________________ >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 hhawkins <@t> utmb.edu Thu May 6 16:56:58 2004 From: hhawkins <@t> utmb.edu (Hawkins, Hal K.) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] lymph nodes Message-ID: <8D6F233E2A5D574B929F3944F3316FD019AD88@EXCH2K3.utmb.edu> My preference (mainly for cutting tough samples of skin) is the Weck prep razor blade -- very sharp, with a well-protected opposite edge. Hal Hawkins, UTMB Galveston -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Smith, Allen Sent: Thursday, May 06, 2004 11:39 AM To: Marshall Terry Dr, Consultant Histopathologist Cc: histonet@pathology.swmed.edu Subject: RE: [Histonet] lymph nodes Most drugstores still carry old fashioned single edge razor blades which have a metal strip folded over the dull edge. They are much easier to dissect with than the disposable microtome blades. I use them for block trimming. 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 Marshall Terry Dr,Consultant Histopathologist Sent: Wednesday, May 05, 2004 11:37 AM To: Mike Bromley; Histonet (E-mail) Subject: RE: [Histonet] lymph nodes I find for this sort of thing, disposable microtome blades are ideal. Holders are available, but I neither use nor recommend them. 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: Mike Bromley [mailto:M.Bromley@dgri.scot.nhs.uk] Sent: 05 May 2004 16:26 To: Histonet (E-mail) Subject: [Histonet] lymph nodes Hi Everyone I am after a very thin blade to slice unfixed lymph nodes, does anyone have any ideas? Ideally some kind of holder for old style razor blades would be the answer. Best Wishes Mike Bromley Chief Biomedical Scientist Pathology Dumfries & Galloway Royal Infirmary Scotland, 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 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) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mrsgbd2001 <@t> yahoo.com Thu May 6 17:17:35 2004 From: mrsgbd2001 <@t> yahoo.com (Gareth Davis) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] cryo rat turbinates Message-ID: <20040506221735.72516.qmail@web13303.mail.yahoo.com> Hello, Thanks to everyone for your suggestions. We have paraffin sections, but we were hoping that cryosections would work, because actually the cryosections looked better. But, we did discover that the slides we used were not as good as SuperFrost Plus. We were told this by the vendor who sold us the slides, after she convinced us that they were just as good. Oh well, you live and learn, it can be expensive - in money and work - but, we will not use those slides anymore. Thanks, I will try the acetone/ethanol fixation on the cryosections I have, but I'm expecting good results. The CryoJane looks interesting, but at the moment we are using another facilities cryostat, so we will have to wait until we have our own cryostat. Thanks again, Gareth Davis Research Assistant Tennessee State University --------------------------------- Do you Yahoo!? Win a $20,000 Career Makeover at Yahoo! HotJobs From bills <@t> icpmr.wsahs.nsw.gov.au Thu May 6 17:39:42 2004 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] lymph nodes In-Reply-To: <8D6F233E2A5D574B929F3944F3316FD019AD88@wsahs.nsw.gov.au> Message-ID: <001001c433bb$06adf060$e287080a@wsahs.nsw.gov.au> In our laboratory, as in many Australian laboratories, we use Feather Disposable Trimming Knives (Cat #130 in a blade holder Cat# FS130S). These are very sharp and easy to use. Specimen dissection is performed using these blades for ALL specimens. They prove to be very beneficial as they allow those performing specimen dissection to cut reasonably thin slices (although sometimes to large in other dimensions) from all the specimens we receive. Even when performing Frozen Sections on fresh lymph nodes (our normal triage procedure) we find the registrar is able to cut reasonably thin blocks of tissue, which then freeze rapidly and evenly. Being so far away from everywhere else in the world we find that cost is a factor but these knives prove to be cost effective as they last longer than scalpels and hand sharpening is time consuming. 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 Hawkins, Hal K. Sent: Friday, 07 May 2004 7:57 AM To: Smith, Allen; Marshall Terry Dr, Consultant Histopathologist Cc: histonet@pathology.swmed.edu Subject: RE: [Histonet] lymph nodes My preference (mainly for cutting tough samples of skin) is the Weck prep razor blade -- very sharp, with a well-protected opposite edge. Hal Hawkins, UTMB Galveston -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Smith, Allen Sent: Thursday, May 06, 2004 11:39 AM To: Marshall Terry Dr, Consultant Histopathologist Cc: histonet@pathology.swmed.edu Subject: RE: [Histonet] lymph nodes Most drugstores still carry old fashioned single edge razor blades which have a metal strip folded over the dull edge. They are much easier to dissect with than the disposable microtome blades. I use them for block trimming. 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 Marshall Terry Dr,Consultant Histopathologist Sent: Wednesday, May 05, 2004 11:37 AM To: Mike Bromley; Histonet (E-mail) Subject: RE: [Histonet] lymph nodes I find for this sort of thing, disposable microtome blades are ideal. Holders are available, but I neither use nor recommend them. 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: Mike Bromley [mailto:M.Bromley@dgri.scot.nhs.uk] Sent: 05 May 2004 16:26 To: Histonet (E-mail) Subject: [Histonet] lymph nodes Hi Everyone I am after a very thin blade to slice unfixed lymph nodes, does anyone have any ideas? Ideally some kind of holder for old style razor blades would be the answer. Best Wishes Mike Bromley Chief Biomedical Scientist Pathology Dumfries & Galloway Royal Infirmary Scotland, 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 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) _______________________________________________ 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 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 KAELPERS <@t> aol.com Thu May 6 22:04:27 2004 From: KAELPERS <@t> aol.com (KAELPERS@aol.com) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] AO 820 microtome questions Message-ID: <75.28d2ce5b.2dcc56bb@aol.com> We have an AO 820 microtome manual that I would be happy to copy and mail to you. It is correct that the blade holder is permanent and the only way to adjust the angle is with the block holder. Email me an address and I will mail you a copy of the manual. I can honestly say that we have purchased 4 newer microtomes in the last 5 years and we still have 2 AO microtomes in the lab, the 2 best microtomist in my lab have been on the AO. Cheers! lge From KAELPERS <@t> aol.com Thu May 6 22:24:00 2004 From: KAELPERS <@t> aol.com (KAELPERS@aol.com) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] cutting rat pancreas Message-ID: <130.2e9c65e2.2dcc5b50@aol.com> Pancreas tissue is always a problem...we have tried cutting larger sections, blocking them with other tissue, and they always seem to be a lot drier. We just block them with intestines and let them soak on ice water to soften. Also you can soak them in a glycerin solution this tends to soften the tissue for good sectioning.....our techs swear by it. lge From anissachoi <@t> hotmail.com Thu May 6 23:22:04 2004 From: anissachoi <@t> hotmail.com (Anissa Choi) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Negative control for immuno slides Message-ID: We do our negative reagent control by omitting the primary antibody. The number of negative slides we run depends on the pre-treatment of the antibodies requested on the case. We run one negative slide for each method i.e. no pre-treatment, protease and pressure cook. Are we doing more negative control slides than what is required ? I know some people just do one negative slide per case. Any comments on the proper way of running negative immuno controls? Thanks in advance for your inputs ! Anissa Choi Burnaby Hospital Burnaby, B.C. Canada _________________________________________________________________ MSN Premium includes powerful parental controls and get 2 months FREE* http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines From mab70 <@t> medschl.cam.ac.uk Fri May 7 02:16:41 2004 From: mab70 <@t> medschl.cam.ac.uk (Margaret Blount) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] cutting rat pancreas Message-ID: <6C21D947F7189448AB447C57BF2992B4025F16AE@mius.medlan.cam.ac.uk> You mention a glycerin solution, what concentration of glycerin? All these tips are very helpful. Thanks Margaret Margaret Blount Chief Technician Clinical Biochemistry University of Cambridge Addenbrooke's Hospital Hills Road Cambridge CB2 2QR -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of KAELPERS@aol.com Sent: Friday, May 07, 2004 4:24 AM To: pstefanova@sten.sunnybrook.utoronto.ca; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] cutting rat pancreas Pancreas tissue is always a problem...we have tried cutting larger sections, blocking them with other tissue, and they always seem to be a lot drier. We just block them with intestines and let them soak on ice water to soften. Also you can soak them in a glycerin solution this tends to soften the tissue for good sectioning.....our techs swear by it. lge _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Stephen.Eyres <@t> sanofi-synthelabo.com Fri May 7 03:02:59 2004 From: Stephen.Eyres <@t> sanofi-synthelabo.com (Stephen.Eyres@sanofi-synthelabo.com) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Old farts reminiscing (was Polyester wax (with earliercitations)) Message-ID: Hi Kemlo, Ooohhh, I've got really bad memories of Sheffield. Leicester used to send techs on the advanced histology course there. The Nurses home on the outskirts was where we stayed. After one very good night our with the local techs, who felt obliged to show us all the good pubs, I went for breakfast the next morning and was met by a pan full of tripe and onions, with a similar one next to it full of black pudding- and not a plastic bag in sight!! Some hopsitality. Did anyone else hand process and double embed eyes through celloidin in 1% methyl benzoate? The first time I was asked to do this procedure, I didn't realise you had to use polyproplylene pots and just a clear plastic one. Came in next morning to find two eyballs sitting on a shelf in glistening goo staring at me. I couldn't help laughing, but that soon changed the the mistake was reported, Having been out of the NHS for 24 yrs, I don't know if Lab Head and Consultant Pathologists have mellowed any, but they were pretty fiece in my day when they wanted to be, and some when they didn't need to be, and my failings were pointed out in no uncertain terms. On very rare occasions, we had a leg amputation brought to the lab fresh from a case of osteosarcoma. We took great delight in scaring juniors to death, by asking for help and then running a scaple blade over the nerves. No-one actually fainted when the toes curled up, but it was a wonder. Cheers Steve "Kemlo Rogerson" To: Sent by: cc: histonet-bounces@lists.utsouth Subject: RE: [Histonet] Old farts reminiscing (was Polyester wax (with western.edu earliercitations)) 06/05/2004 17:11 Anyway, I miss the older generation of histotechnicians (as we used to call them over here). My first encounter was with one Des Brady in Sheffield. He was great. If you asked for a special stain, he would say "do you want it positive or negative boss?" That's what I call skill:-) Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Fri May 7 05:06:38 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Old farts reminiscing (was Polyester wax (withearliercitations)) Message-ID: Stephen says: "I don't know if Lab Head and Consultant Pathologists have mellowed any, ..." Of course. Now we are all sweet and loveable. I didn't know it had been otherwise. Dr Terry (call me Teddybear ) 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: Stephen.Eyres@sanofi-synthelabo.com [mailto:Stephen.Eyres@sanofi-synthelabo.com] Sent: 07 May 2004 09:03 To: Kemlo Rogerson; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Old farts reminiscing (was Polyester wax (withearliercitations)) Hi Kemlo, Ooohhh, I've got really bad memories of Sheffield. Leicester used to send techs on the advanced histology course there. The Nurses home on the outskirts was where we stayed. After one very good night our with the local techs, who felt obliged to show us all the good pubs, I went for breakfast the next morning and was met by a pan full of tripe and onions, with a similar one next to it full of black pudding- and not a plastic bag in sight!! Some hopsitality. Did anyone else hand process and double embed eyes through celloidin in 1% methyl benzoate? The first time I was asked to do this procedure, I didn't realise you had to use polyproplylene pots and just a clear plastic one. Came in next morning to find two eyballs sitting on a shelf in glistening goo staring at me. I couldn't help laughing, but that soon changed the the mistake was reported, Having been out of the NHS for 24 yrs, I don't know if Lab Head and Consultant Pathologists have mellowed any, but they were pretty fiece in my day when they wanted to be, and some when they didn't need to be, and my failings were pointed out in no uncertain terms. On very rare occasions, we had a leg amputation brought to the lab fresh from a case of osteosarcoma. We took great delight in scaring juniors to death, by asking for help and then running a scaple blade over the nerves. No-one actually fainted when the toes curled up, but it was a wonder. Cheers Steve "Kemlo Rogerson" To: Sent by: cc: histonet-bounces@lists.utsouth Subject: RE: [Histonet] Old farts reminiscing (was Polyester wax (with western.edu earliercitations)) 06/05/2004 17:11 Anyway, I miss the older generation of histotechnicians (as we used to call them over here). My first encounter was with one Des Brady in Sheffield. He was great. If you asked for a special stain, he would say "do you want it positive or negative boss?" That's what I call skill:-) Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. _______________________________________________ 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 Stephen.Eyres <@t> sanofi-synthelabo.com Fri May 7 06:13:50 2004 From: Stephen.Eyres <@t> sanofi-synthelabo.com (Stephen.Eyres@sanofi-synthelabo.com) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Old farts reminiscing (was Polyester wax (withearliercitations)) Message-ID: Hi Terry, But is that because, like us old timers, you're getting old and senile? cheers Steve "Marshall Terry Dr, Consultant To: Stephen Eyres/GB-ALNWICK/RESEARCH/SANOFI@Research Histopathologist" "Kemlo Rogerson" gen.nhs.uk> cc: Subject: RE: [Histonet] Old farts reminiscing (was Polyester wax 07/05/2004 11:06 (withearliercitations)) Stephen says: "I don't know if Lab Head and Consultant Pathologists have mellowed any, ..." Of course. Now we are all sweet and loveable. I didn't know it had been otherwise. Dr Terry (call me Teddybear ) 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: Stephen.Eyres@sanofi-synthelabo.com [mailto:Stephen.Eyres@sanofi-synthelabo.com] Sent: 07 May 2004 09:03 To: Kemlo Rogerson; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Old farts reminiscing (was Polyester wax (withearliercitations)) Hi Kemlo, Ooohhh, I've got really bad memories of Sheffield. Leicester used to send techs on the advanced histology course there. The Nurses home on the outskirts was where we stayed. After one very good night our with the local techs, who felt obliged to show us all the good pubs, I went for breakfast the next morning and was met by a pan full of tripe and onions, with a similar one next to it full of black pudding- and not a plastic bag in sight!! Some hopsitality. Did anyone else hand process and double embed eyes through celloidin in 1% methyl benzoate? The first time I was asked to do this procedure, I didn't realise you had to use polyproplylene pots and just a clear plastic one. Came in next morning to find two eyballs sitting on a shelf in glistening goo staring at me. I couldn't help laughing, but that soon changed the the mistake was reported, Having been out of the NHS for 24 yrs, I don't know if Lab Head and Consultant Pathologists have mellowed any, but they were pretty fiece in my day when they wanted to be, and some when they didn't need to be, and my failings were pointed out in no uncertain terms. On very rare occasions, we had a leg amputation brought to the lab fresh from a case of osteosarcoma. We took great delight in scaring juniors to death, by asking for help and then running a scaple blade over the nerves. No-one actually fainted when the toes curled up, but it was a wonder. Cheers Steve "Kemlo Rogerson" To: Sent by: cc: histonet-bounces@lists.utsouth Subject: RE: [Histonet] Old farts reminiscing (was Polyester wax (with western.edu earliercitations)) 06/05/2004 17:11 Anyway, I miss the older generation of histotechnicians (as we used to call them over here). My first encounter was with one Des Brady in Sheffield. He was great. If you asked for a special stain, he would say "do you want it positive or negative boss?" That's what I call skill:-) Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. _______________________________________________ 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 celebrej <@t> HHSC.CA Fri May 7 06:20:03 2004 From: celebrej <@t> HHSC.CA (Celebre Julia) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Control tissue needed Message-ID: <3AADFB88753AD31189C100902786B91C0E41C9A3@hch_nt_exchange.hhsc.ca> Happy Friday to all... Is anyone out there in Histoland willing to share some much needed control blocks for our suddenly popular Wade-Fite special stain for Leprosy? Or, does anyone know of a resource for hard to find control blocks? For years we would always run a TB control thru our Wade-Fite stain up until we got a hold of 20 slides positive for Leprosy which are now depleted, no help to our eager pathologists......... Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext 46145 email: celebrej@hhsc.ca This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From Kevin.Randall <@t> astrazeneca.com Fri May 7 06:19:35 2004 From: Kevin.Randall <@t> astrazeneca.com (Randall, Kevin J) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] RE: lymph nodes Message-ID: I prefer to use the Skin Graft blade (SG3) from Swann-Morton (http://www.swann-morton.com/blades3.html). These fit on a standard No.3 scalpel handle and are extremely sharp. I use them for cutting very thin slices of fresh rat kidney and they perform far better than microtome or razor blades. Regards Kevin Randall AstraZeneca Alderley Park UK From bhewlett <@t> cogeco.ca Fri May 7 07:40:14 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Negative control for immuno slides References: Message-ID: <002201c43430$76065ea0$6500a8c0@mainbox> Anissa, The ideal way of running negative reagent controls is to substitute an Ig isotype-matched Monoclonal antibody, directed against an antigen not normally found in human material, at the same concentration as your primary antibody in the same diluent. Alternately, including in your panel of antibodies another monoclonal primary antibody of the same Ig isotype, using the same pre-treatment but with a totally different distribution of reactivity, is acceptable as a control. Failing that, simply applying the primary antibody diluent, as you are doing, is acceptable according to NCCLS guidelines. For each case, following each method variation, the negative reagent control for each primary must be applied as you are doing. However, should you use several monoclonal antibodies on a case, of the same Ig isotype with the same pre-treatment, then only one negative reagent control for those would be required. Bryan Hewlett Technical Consultant QMP-LS, Ontario, Canada ----- Original Message ----- From: "Anissa Choi" To: Sent: Friday, May 07, 2004 12:22 AM Subject: [Histonet] Negative control for immuno slides > We do our negative reagent control by omitting the primary antibody. The > number of negative > slides we run depends on the pre-treatment of the antibodies requested on > the case. We run one > negative slide for each method i.e. no pre-treatment, protease and pressure > cook. Are we doing > more negative control slides than what is required ? I know some people just > do one negative > slide per case. Any comments on the proper way of running negative immuno > controls? Thanks > in advance for your inputs ! > > > Anissa Choi > Burnaby Hospital > Burnaby, B.C. > Canada > > _________________________________________________________________ > MSN Premium includes powerful parental controls and get 2 months FREE* > http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=htt p://hotmail.com/enca&HL=Market_MSNIS_Taglines > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From kemlo <@t> tiscali.co.uk Fri May 7 09:02:29 2004 From: kemlo <@t> tiscali.co.uk (Kemlo) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Old farts reminiscing (was Polyester wax (withearliercitations)) In-Reply-To: Message-ID: <4076DBCE0005EB4D@mk-cpfrontend-3.mail.uk.tiscali.com> Teddy bear! __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From funderwood <@t> mcohio.org Fri May 7 09:08:34 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Control tissue needed Message-ID: Hi Julia. American Master Tech Scientific has leprosy control slides 1.800.860.4073 or americanhistology.com Have a great weekend! Fred >>> Celebre Julia 05/07/04 07:20AM >>> Happy Friday to all... Is anyone out there in Histoland willing to share some much needed control blocks for our suddenly popular Wade-Fite special stain for Leprosy? Or, does anyone know of a resource for hard to find control blocks? For years we would always run a TB control thru our Wade-Fite stain up until we got a hold of 20 slides positive for Leprosy which are now depleted, no help to our eager pathologists......... Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext 46145 email: celebrej@hhsc.ca This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Stephen.Eyres <@t> sanofi-synthelabo.com Fri May 7 09:11:04 2004 From: Stephen.Eyres <@t> sanofi-synthelabo.com (Stephen.Eyres@sanofi-synthelabo.com) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Old farts reminiscing (was Polyester wax (withearliercitations)) Message-ID: Bet you could think of some more traditional words!! "Kemlo" "Marshall Terry Dr, Consultant Histopathologist" cc: histonet@lists.utsouthwestern.edu 07/05/2004 15:02 Subject: RE: [Histonet] Old farts reminiscing (was Polyester wax Please respond (withearliercitations)) to kemlo Teddy bear! __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From JMcCormi <@t> schosp.org Fri May 7 09:21:52 2004 From: JMcCormi <@t> schosp.org (McCormick, James) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Old farts reminiscing (was Polyester wax (withearl iercitations)) Message-ID: <229A3566B9F0D311826E00D0B7441D7905E2FCE5@swedish_nt1.schosp.org> Not all bad.....to be traditional we might suggest the use of "antique and flatulent!"....JBMc -----Original Message----- From: Stephen.Eyres@sanofi-synthelabo.com [mailto:Stephen.Eyres@sanofi-synthelabo.com] Sent: Friday, May 07, 2004 9:11 AM To: kemlo@tiscali.co.uk Cc: histonet@lists.utsouthwestern.edu; Marshall Terry Dr, Consultant Histopathologist Subject: RE: [Histonet] Old farts reminiscing (was Polyester wax (withearliercitations)) Bet you could think of some more traditional words!! "Kemlo" "Marshall Terry Dr, Consultant Histopathologist" cc: histonet@lists.utsouthwestern.edu 07/05/2004 15:02 Subject: RE: [Histonet] Old farts reminiscing (was Polyester wax Please respond (withearliercitations)) to kemlo Teddy bear! __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM _______________________________________________ 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 MinHan.Tan <@t> vai.org Fri May 7 09:52:14 2004 From: MinHan.Tan <@t> vai.org (Tan, MinHan) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] What's cooking...? Only slides Message-ID: <74D0F0AB07F2E647A02D839ED79520F9B486E6@VAIEXCH02.vai.org> Good morning, We are planning to purchase a pressure cooker immediately for a particularly troublesome antigen Unfortunately, no one here has used one before for antigen retrieval, and we have a couple of questions (which may seem rather silly). a. Most posts on Histonet seem to have used either the Nordicware cooker ($45) or the Biocare decloaker ($?). Does a 'decloaker' work differently from a pressure cooker? b. Pressure cooking of slides - most protocols seems to range from 3 min - 5 min. I haven't done much cooking myself (thanks to a wonderful wife), but others in my lab assure me that it takes time for a cooker to build up pressure. In steam retrieval, I usually check to ensure that the buffer is at 95 deg C before starting AR. Is there some kind of equivalent for this in 'pressure cooking'? Thanks! (and apologies for my culinary incompetence) Min-Han Tan 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 laurie.colbert <@t> huntingtonhospital.com Fri May 7 09:56:35 2004 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] archives Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5BEEF@EXCHANGE1.huntingtonhospital.com> I know this has been asked many times, but how do I access the archives (I'm specifically looking for the talk on EGFR testing). Thanks! Laurie Colbert From gcallis <@t> montana.edu Fri May 7 10:00:15 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Negative control for immuno slides In-Reply-To: Message-ID: <3.0.6.32.20040507090015.00c270e8@gemini.msu.montana.edu> Anissa, Removal of the primary is NOT a negative control, you should replace the primary antibody with the immunoglobulin of host of the primary, ie. if you work with Rat antiMouse antibody, IgG2a the negative control is either Rat IgG which contains IgG2a isotype or use Rat IgG 2a and at the same concentration as the primary antibody. You must have primary host IgG in the system in order to know IF this immunoglobulin is crossreacting to tissue being stained. The negative control section must be treated exactly as you treat the unknown tissue, including all steps of your IHC protocol. OR you can use an irrelevant antibody of same IgG isotype that does not recognize you antigen OR you can use a transgenic knockout animal known to NOT express the antigen you are interested in staining. However, on the latter, knockout animal, we have found the host IgG sometimes does cross react, so we still do the isotype matched control or immunoglobulin control. We do NOT use normal serums as they can contain nonspecific antibodies - presenting background problems. Jackson Immunoresearch has a wonderful array of different species immunoglobulins that are very inexpensive and clean. If you have several antibodies with same host of primary, and at same antibody concentrations, you can get away with one control for many antibodies. DAKO manual by Boenisch discussion on this is very clear, and a good read, go to DAKO website, click on support and read the pdf file on this. Removal of primary is considered a NULL control, and has its use when you want to do a reagents test for background. At 09:22 PM 5/6/2004 -0700, you wrote: >We do our negative reagent control by omitting the primary antibody. The >number of negative >slides we run depends on the pre-treatment of the antibodies requested on >the case. We run one >negative slide for each method i.e. no pre-treatment, protease and pressure >cook. Are we doing >more negative control slides than what is required ? I know some people just >do one negative >slide per case. Any comments on the proper way of running negative immuno >controls? Thanks >in advance for your inputs ! > > >Anissa Choi >Burnaby Hospital >Burnaby, B.C. >Canada > >_________________________________________________________________ >MSN Premium includes powerful parental controls and get 2 months FREE* >http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=ht tp://hotmail.com/enca&HL=Market_MSNIS_Taglines > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From lpjones <@t> srhs-pa.org Fri May 7 09:58:41 2004 From: lpjones <@t> srhs-pa.org (Jones, Laura) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Pathology Waste From Autopsies Message-ID: We are wondering what everyone is using to dispose of body parts removed during autopsy. We currently used any old box lined with double red bags, but clearly labeled with biohazard stickers and "BODY PARTS", then the whole box is placed into another red biohazard bag. Are there special boxes for this purpose, such as the "Burn Boxes" we have found in catalogs? 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 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 message. From lpjones <@t> srhs-pa.org Thu May 6 14:45:13 2004 From: lpjones <@t> srhs-pa.org (Jones, Laura) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Pathology Waste From Autopsies Message-ID: We are wondering what everyone is using to dispose of body parts removed during autopsy. We currently used any old box lined with double red bags, but clearly labeled with biohazard stickers and "BODY PARTS", then the whole box is placed into another red biohazard bag. Are there special boxes for this purpose, such as the "Burn Boxes" we have found in catalogs? 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 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 message. From lpjones <@t> srhs-pa.org Fri May 7 09:51:08 2004 From: lpjones <@t> srhs-pa.org (Jones, Laura) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] Pathology Waste From Autopsies Message-ID: We are wondering what everyone is using to dispose of body parts removed during autopsy. We currently used any old box lined with double red bags, but clearly labeled with biohazard stickers and "BODY PARTS", then the whole box is placed into another red biohazard bag. Are there special boxes for this purpose, such as the "Burn Boxes" we have found in catalogs? 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 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 message. From ssalesky <@t> LowellGeneral.org Fri May 7 10:52:45 2004 From: ssalesky <@t> LowellGeneral.org (Shawn Salesky) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] RE: Histonet Digest, Vol 6, Issue 9 Message-ID: <7F5DFD606D34D6118B520008C756AA8701E105C7@mxtwo.lowellgeneral.org> Hello, I find Disposable microtome blade to be perfect for this also. Low profile blades DO NOT work as well as the regular blades. (Only because there is not enough to grip properly) As an example http://www.mopec.com/chapter3/ch3_Blades_Blade_Holders.htm Shawn Hi Everyone I am after a very thin blade to slice unfixed lymph nodes, does anyone have any ideas? Ideally some kind of holder for old style razor blades would be the answer. Best Wishes Mike Bromley Chief Biomedical Scientist Pathology Dumfries & Galloway Royal Infirmary Scotland, UK From powell_sa <@t> Mercer.edu Fri May 7 11:12:25 2004 From: powell_sa <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:22:55 2005 Subject: [Histonet] archives References: <0BE6ADFAE4E7E04496BF21ABD346628001C5BEEF@EXCHANGE1.huntingtonhospital.com> Message-ID: <001b01c4344e$15fdbb20$e3f2acd1@powellsa1> Hi Laurie, go to www.histosearch.com and click on the drop down box on the left, choose histonet archives, then type in the subject for which you are searching in the box to the right, then click search. All of the previous emails concerning your subject will be displayed for you. Shirley Powell spowell@histosearch.com ----- Original Message ----- From: "Laurie Colbert" To: Sent: Friday, May 07, 2004 10:56 AM Subject: [Histonet] archives I know this has been asked many times, but how do I access the archives (I'm specifically looking for the talk on EGFR testing). Thanks! Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet = From laurie.colbert <@t> huntingtonhospital.com Fri May 7 11:24:29 2004 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Archives Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5BEF3@EXCHANGE1.huntingtonhospital.com> Thanks to everyone who sent me info on accessing the archives - got it!! Laurie Colbert From Terry.Marshall <@t> rothgen.nhs.uk Fri May 7 11:29:25 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] RE: lymph nodes Message-ID: Yes, these too are good. However, I don't like the heavy handle and light blade, a problem I also conceive with the Feather blade holders and the dreaded PM40 knife, an abomination if ever there was one. The problem is lack of feel of where the blade is. With the heavy handle you can only feel where the handle is. I think this is plain dangerous, far more so than holding the blade in your fingers, which in my view, is not dangerous at all. In point of fact, all the things that have been mentioned are satisfactory. Confession: I have my own ham knife and sharpener, lately, a ceramic sharpener. These are mine and only mine. Nobody touches them. I keep it sharp - much sharper than a scalpel. The length of the blade allows a proper slicing cut, and I use this for EVERYTHING - no exception. Changing tack slightly, I am tempted to cut some holes in the blade, like the latest generation of Japanese and cheese knives. This apparently lessens the suction or drag on the blade. 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: Randall, Kevin J [mailto:Kevin.Randall@astrazeneca.com] Sent: 07 May 2004 12:20 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] RE: lymph nodes I prefer to use the Skin Graft blade (SG3) from Swann-Morton (http://www.swann-morton.com/blades3.html). These fit on a standard No.3 scalpel handle and are extremely sharp. I use them for cutting very thin slices of fresh rat kidney and they perform far better than microtome or razor blades. Regards Kevin Randall AstraZeneca Alderley Park UK _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mhorne <@t> upei.ca Fri May 7 11:37:47 2004 From: mhorne <@t> upei.ca (Margaret Horne) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] enzymes in the nucleus Message-ID: <409B912A.28899.121D367@localhost> Hi , We have been staining for COX 1 and COX 2 in eye and brain and have been noticing specific staining in the nucleus but not the nucleolus. We are wondering if anybody has noticed staining of enzymes in the nucleus as well as in the cytoplasm. Thanks , Margaret Margaret Horne , Histology Teaching Assistant, Dept. of B.SC., Atlantic Veterinary College, U.P.E.I., 550 University Ave., Charlottetown, P.E.I., C1A 4P3 Canada From gcallis <@t> montana.edu Fri May 7 12:02:23 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Knife angle, old 820 disposable holder and block angle? further comments In-Reply-To: <75.28d2ce5b.2dcc56bb@aol.com> Message-ID: <3.0.6.32.20040507110223.00bf7a18@gemini.msu.montana.edu> Felt the need for further comment about blade angle with older AO 820 microtome. You wrote: "It is correct that the blade holder is permanent and the only way to adjust the angle is with the block holder". You do not adjust the blade angle i.e the cutting angle at the block holder - block holder adjustment (the x/y axis adjusment) only provides is orientation of block face to the blade which is set at its cutting angle. Once block is trimmed and ready for sectioning, you are locked into using the cutting angle - determined by blade holder and bevel angle ON disposable blade. With today's tissue cassettes, block face adjustment has become blessedly minimal - we only readjust block face for some block recuts but we do change holder adjustments to accomodate the blade being used. We found using old AO 820 blade holder that some brands of knives are slightly different due to how they were manufacturered/treated. Some blades just work better than others in this holder, and you may have have to try various brands. Ask for samples if you experience too much difficulty with any one disposable. blade. A significant reason why blade holders are now totally adjustable is probably because the old holder WAS a permanent angle. Microtomists still need to refine cutting angles for any given blade use. There have been long discussions about this on Histonet, check out archives. One can use the 820 standard knife holder (for C profile steel knives), buy a disposable blade insert to achieve fine tweaking of cutting angle. This may be important IF one really dislikes or cannot work with old 820 holder. Have a good weekend after this long (whew!) lecture 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 kmerriam2003 <@t> yahoo.com Fri May 7 12:25:56 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] article about Pathology in boston globe Message-ID: <20040507172556.63479.qmail@web51002.mail.yahoo.com> Thought you all would enjoy this: http://www.boston.com/news/globe/health_science/articles/2004/05/04/pathology_playing_god_in_the_laboratory/ Kim Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Win a $20,000 Career Makeover at Yahoo! HotJobs From MDiCarlo <@t> KaleidaHealth.Org Fri May 7 12:21:28 2004 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] acetyl-cholinesterase Message-ID: Histonetters, Is anyone out there using acetyl-cholinesterase for aganglionosis? If so where where do you purchase your kit? Thanks for your help. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to 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, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. From sladd <@t> hsc.usf.edu Fri May 7 12:41:48 2004 From: sladd <@t> hsc.usf.edu (S Ladd) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Knife angle, old 820 disposable holder and block angle?further comments In-Reply-To: <3.0.6.32.20040507110223.00bf7a18@gemini.msu.montana.edu> Message-ID: I have an old AO 820 microtome and I have the non-adjustable blade holder. I use DuraEdge high profile blades (#7310)--1-800-253-2768 and they work great (even with 4 micron sections)!! I couldn't get the Leica blades to work at all. I have also tried an Reichert insert-type holder in my adjustable steel knife holder but that didn't seem to work as well either. Again, a lot of this is just personal preference. I would try some different types of blades before you give up on the non-adjustable blade holder. Sharron -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gayle Callis Sent: Friday, May 07, 2004 1:02 PM To: KAELPERS@aol.com; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Knife angle, old 820 disposable holder and block angle?further comments Felt the need for further comment about blade angle with older AO 820 microtome. You wrote: "It is correct that the blade holder is permanent and the only way to adjust the angle is with the block holder". You do not adjust the blade angle i.e the cutting angle at the block holder - block holder adjustment (the x/y axis adjusment) only provides is orientation of block face to the blade which is set at its cutting angle. Once block is trimmed and ready for sectioning, you are locked into using the cutting angle - determined by blade holder and bevel angle ON disposable blade. With today's tissue cassettes, block face adjustment has become blessedly minimal - we only readjust block face for some block recuts but we do change holder adjustments to accomodate the blade being used. We found using old AO 820 blade holder that some brands of knives are slightly different due to how they were manufacturered/treated. Some blades just work better than others in this holder, and you may have have to try various brands. Ask for samples if you experience too much difficulty with any one disposable. blade. A significant reason why blade holders are now totally adjustable is probably because the old holder WAS a permanent angle. Microtomists still need to refine cutting angles for any given blade use. There have been long discussions about this on Histonet, check out archives. One can use the 820 standard knife holder (for C profile steel knives), buy a disposable blade insert to achieve fine tweaking of cutting angle. This may be important IF one really dislikes or cannot work with old 820 holder. Have a good weekend after this long (whew!) lecture 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 MDiCarlo <@t> KaleidaHealth.Org Fri May 7 13:13:23 2004 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Knife angle, old 820 disposable holder and block a ngle?further comments Message-ID: I use Accu-Edge high profile blades and cut at 4 microns and have great sections using the non adjustable knife holder for the old AO microtomes. Peggy DiCarlo -----Original Message----- From: S Ladd [mailto:sladd@hsc.usf.edu] Sent: Friday, May 07, 2004 13:42 To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Knife angle, old 820 disposable holder and block angle?further comments I have an old AO 820 microtome and I have the non-adjustable blade holder. I use DuraEdge high profile blades (#7310)--1-800-253-2768 and they work great (even with 4 micron sections)!! I couldn't get the Leica blades to work at all. I have also tried an Reichert insert-type holder in my adjustable steel knife holder but that didn't seem to work as well either. Again, a lot of this is just personal preference. I would try some different types of blades before you give up on the non-adjustable blade holder. Sharron -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gayle Callis Sent: Friday, May 07, 2004 1:02 PM To: KAELPERS@aol.com; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Knife angle, old 820 disposable holder and block angle?further comments Felt the need for further comment about blade angle with older AO 820 microtome. You wrote: "It is correct that the blade holder is permanent and the only way to adjust the angle is with the block holder". You do not adjust the blade angle i.e the cutting angle at the block holder - block holder adjustment (the x/y axis adjusment) only provides is orientation of block face to the blade which is set at its cutting angle. Once block is trimmed and ready for sectioning, you are locked into using the cutting angle - determined by blade holder and bevel angle ON disposable blade. With today's tissue cassettes, block face adjustment has become blessedly minimal - we only readjust block face for some block recuts but we do change holder adjustments to accomodate the blade being used. We found using old AO 820 blade holder that some brands of knives are slightly different due to how they were manufacturered/treated. Some blades just work better than others in this holder, and you may have have to try various brands. Ask for samples if you experience too much difficulty with any one disposable. blade. A significant reason why blade holders are now totally adjustable is probably because the old holder WAS a permanent angle. Microtomists still need to refine cutting angles for any given blade use. There have been long discussions about this on Histonet, check out archives. One can use the 820 standard knife holder (for C profile steel knives), buy a disposable blade insert to achieve fine tweaking of cutting angle. This may be important IF one really dislikes or cannot work with old 820 holder. Have a good weekend after this long (whew!) lecture 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to 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, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. From JPCOLEMA <@t> sentara.com Fri May 7 13:18:53 2004 From: JPCOLEMA <@t> sentara.com (JOHN COLEMAN) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] not wine, PTAH!! Message-ID: Since we're Histotechs shouldn't we be like a fine shelf-ripened Phosphotungstic Acid Hematoxylin unfiltered and naturally aged without the addition of potassium permanganate? Wine is so cliche. From funderwood <@t> mcohio.org Fri May 7 13:26:06 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] not wine, PTAH!! Message-ID: Mondays I tend to feel like dried out picric acid. Chalky, flakey and kind of volatile. Fred >>> "JOHN COLEMAN" 05/07/04 02:18PM >>> Since we're Histotechs shouldn't we be like a fine shelf-ripened Phosphotungstic Acid Hematoxylin unfiltered and naturally aged without the addition of potassium permanganate? Wine is so cliche. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hhawkins <@t> utmb.edu Fri May 7 13:42:14 2004 From: hhawkins <@t> utmb.edu (Hawkins, Hal K.) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] RE: Weck prep razor blade Message-ID: <8D6F233E2A5D574B929F3944F3316FD01A581E@EXCH2K3.utmb.edu> They are officially the "Pilling Weck Surgical Wecprep," Pilling Weck Surgical, Inc., Fort Washington, PA, 19034, Cat. No. 450205. http://www.pillingsurgical.com/ They are also available from ems: http://www.emsdiasum.com/microscopy/products/preparation/blades.aspx?mm= 8 -----Original Message----- From: Denise [mailto:denise@pclv.net] Sent: Thursday, May 06, 2004 10:56 PM To: Hawkins, Hal K. Subject: Weck prep razor blade Hal, Where does one find the "the Weck prep razor blade"? denise@pclv.net From JPCOLEMA <@t> sentara.com Fri May 7 13:48:05 2004 From: JPCOLEMA <@t> sentara.com (JOHN COLEMAN) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] AFB= bad ctrl for Fite, Negative controls, decloaker, egfr Message-ID: Using an AFB control for a Fite stain is a dangerous game! It is quite possible for the control to look positive when the capsule of the bacteria in question on the test slide is present and dissolved and the slide therefore looks negative! For Negative controls, in addition to matching all the rest of the protocol like you already are doing, you must at least also apply the same diluent used for the primary antibodies since letting the slide dry out will in most cases alter the stainability of the slide if there were an issue of false positivity gotten from the secondary reagents, retrieval or other ancillary procedures or reagents. Biocare's decloaker is basically a fine tuned lab grade pressure cooker- we use neither. We tried the pressure cooker offered by Cellmarque, and it saved us no time, and did not significantly improve retrieval.What antibody are you having trouble with? I could tell you what we do if it's one we use. I tested 6 EGFRs on 45 breast cancer and 30 colon cancers looking for a percentage that matched that of Dako's kit and I have to say that the kit's clone is the best I've found. The other clones have a 5-10 % staining of all cases and The Dako kit had a yield closer to 50%, which is what the research for the drug claims. We bit the bullet ( price tag) and now order the kit. From Rcartun <@t> harthosp.org Fri May 7 14:06:15 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] AFB= bad ctrl for Fite, Negative controls, decloaker,egfr Message-ID: Dear John: I was interested in your comments regarding EGFR testing. I have been using Zymed's mAb (clone 31G7) for years and it almost always gives more immunoreactivity than the mAb in DakoCytomation's EGFR kit. For the determination of patient eligibility for anti-EGFR therapy, we report the result for the Dako test, but we always run the Zymed mAb in parallel, and sometimes use it to help us determine if a patient's tumor is truly positive (or negative). Richard Cartun >>> "JOHN COLEMAN" 05/07/04 02:48PM >>> Using an AFB control for a Fite stain is a dangerous game! It is quite possible for the control to look positive when the capsule of the bacteria in question on the test slide is present and dissolved and the slide therefore looks negative! For Negative controls, in addition to matching all the rest of the protocol like you already are doing, you must at least also apply the same diluent used for the primary antibodies since letting the slide dry out will in most cases alter the stainability of the slide if there were an issue of false positivity gotten from the secondary reagents, retrieval or other ancillary procedures or reagents. Biocare's decloaker is basically a fine tuned lab grade pressure cooker- we use neither. We tried the pressure cooker offered by Cellmarque, and it saved us no time, and did not significantly improve retrieval.What antibody are you having trouble with? I could tell you what we do if it's one we use. I tested 6 EGFRs on 45 breast cancer and 30 colon cancers looking for a percentage that matched that of Dako's kit and I have to say that the kit's clone is the best I've found. The other clones have a 5-10 % staining of all cases and The Dako kit had a yield closer to 50%, which is what the research for the drug claims. We bit the bullet ( price tag) and now order the kit. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kwittle <@t> jhmi.edu Fri May 7 14:24:49 2004 From: kwittle <@t> jhmi.edu (Karen Wittler) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] I have reviewed Re: Nair to soften nails & Re: Tough Toenails in the archives. Ron Slyter recommend Message-ID: I have reviewed Re: Nair to soften nails & Re: Tough Toenails in the archives. Ron Slyter recommended using Calcium Hydroxide, the active ingredient in "Nair" to soften nails. Is there a procedure, a dilution ? Su, Phy-Huynh recommended using 3-aminopropyltriethoxysilane coated slides to aid in keeping toenail sections on the slide. As I do not have a Histochemical Journal 1986 : 18 , I would appreciate some help with recommendations, or a proceedure. We silanized our slides many years ago but I do not remember how we made our solution. We are prepared to search through archived proceedure manuals, but perhaps input from others will provide more information than our old proceedures. Any help will be well appreciated. Thank you, Karen Wittler, HT(ASCP) Johns Hopkins Hospital Baltimore, MD From weneng <@t> hotmail.com Fri May 7 14:20:55 2004 From: weneng <@t> hotmail.com (Wendy England) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] tissue slice tool Message-ID: Hello, We are using razor blades to slice tissue. I am wondering if there is a special tool that has two blades 2-3 mm apart so that the slice can be made by one cut and slice should be more flat. Anybody has idea where I can find it or it's just not practical? Thanks, Wendy _________________________________________________________________ Watch LIVE baseball games on your computer with MLB.TV, included with MSN Premium! http://join.msn.com/?page=features/mlb&pgmarket=en-us/go/onm00200439ave/direct/01/ From japoteete <@t> saintfrancis.com Fri May 7 15:00:36 2004 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] AFB= bad ctrl for Fite, Negative controls, decloak er,egfr Message-ID: Running the EGFRs in duplicate seems to be a bit of overkill. The kit received FDA approval based on the testing of the clone and the components of the kit. As the Erbitux rep explained to us, the drug is dispensed based upon results from the DAKO EGFR-Dx only. Of course, I suppose this premise is good in theory, but we all know that some Oncologists don't always follow each and every rule. Just my humble opinion. Jacquie Poteete, MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com > -----Original Message----- > From: Richard Cartun [SMTP:Rcartun@harthosp.org] > Sent: Friday, May 07, 2004 2:06 PM > To: histonet@lists.utsouthwestern.edu; JPCOLEMA@sentara.com > Subject: Re: [Histonet] AFB= bad ctrl for Fite, Negative controls, > decloaker,egfr > > Dear John: > > I was interested in your comments regarding EGFR testing. I have been > using Zymed's mAb (clone 31G7) for years and it almost always gives more > immunoreactivity than the mAb in DakoCytomation's EGFR kit. For the > determination of patient eligibility for anti-EGFR therapy, we report > the result for the Dako test, but we always run the Zymed mAb in > parallel, and sometimes use it to help us determine if a patient's tumor > is truly positive (or negative). > > Richard Cartun > > >>> "JOHN COLEMAN" 05/07/04 02:48PM >>> > Using an AFB control for a Fite stain is a dangerous game! It is quite > possible for the control to look positive when the capsule of the > bacteria in question on the test slide is present and dissolved and the > slide therefore looks negative! > > For Negative controls, in addition to matching all the rest of the > protocol like you already are doing, you must at least also apply the > same diluent used for the primary antibodies since letting the slide dry > out will in most cases alter the stainability of the slide if there were > an issue of false positivity gotten from the secondary reagents, > retrieval or other ancillary procedures or reagents. > > Biocare's decloaker is basically a fine tuned lab grade pressure > cooker- we use neither. We tried the pressure cooker offered by > Cellmarque, and it saved us no time, and did not significantly improve > retrieval.What antibody are you having trouble with? I could tell you > what we do if it's one we use. > > I tested 6 EGFRs on 45 breast cancer and 30 colon cancers looking for > a percentage that matched that of Dako's kit and I have to say that the > kit's clone is the best I've found. The other clones have a 5-10 % > staining of all cases and The Dako kit had a yield closer to 50%, which > is what the research for the drug claims. We bit the bullet ( price tag) > and now order the kit. > _______________________________________________ > 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 > > > > ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp From louri_c <@t> hotmail.com Fri May 7 16:03:32 2004 From: louri_c <@t> hotmail.com (Louri) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] RE: Incoming Msg Message-ID: [cid:ffsqqyonsm.bmp] From Luis.Chiriboga <@t> med.nyu.edu Fri May 7 14:42:03 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] article about Pathology in boston globe In-Reply-To: <20040507172556.63479.qmail@web51002.mail.yahoo.com> Message-ID: funny, they don't even mention histologist........like the slides magically appeared -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Kim Merriam Sent: Friday, May 07, 2004 1:26 PM To: Histonet Subject: [Histonet] article about Pathology in boston globe Thought you all would enjoy this: http://www.boston.com/news/globe/health_science/articles/2004/05/04/patholog y_playing_god_in_the_laboratory/ Kim Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Win a $20,000 Career Makeover at Yahoo! HotJobs _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lagehan <@t> hotmail.com Fri May 7 15:13:28 2004 From: lagehan <@t> hotmail.com (LORALEE GEHAN) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] I have reviewed Re: Nair to soften nails & Re: Tough Toenails in the archives. Ron Slyter recommend Message-ID: All we do is put the nail in a blob of nair for at least 24 hours. Then process as usual. After processing you can trim the block at bit and then ice it really well. It cuts beautifully. We get them on a weekly basis and also do special stains on them as well. I think that it is cheaper than Calcium Hydroxide. Hope that helps. Loralee Gehan, HTL Histology Supervisor Mid-Atlantic Pathology Services, Inc. >From: Karen Wittler >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] I have reviewed Re: Nair to soften nails & Re: Tough Toenails in the archives. Ron Slyter recommend >Date: Fri, 07 May 2004 15:24:49 -0400 > > I have reviewed Re: Nair to soften nails & Re: Tough Toenails in the >archives. Ron Slyter recommended using Calcium Hydroxide, the active >ingredient in "Nair" to soften nails. Is there a procedure, a dilution >? > > Su, Phy-Huynh recommended using 3-aminopropyltriethoxysilane coated >slides to aid in keeping toenail sections on the slide. As I do not have >a Histochemical Journal 1986 : 18 , I would appreciate some help with >recommendations, or a proceedure. We silanized our slides many years ago >but I do not remember how we made our solution. > > We are prepared to search through archived proceedure manuals, but >perhaps input from others will provide more information than our old >proceedures. Any help will be well appreciated. > > Thank you, > >Karen Wittler, HT(ASCP) >Johns Hopkins Hospital >Baltimore, MD > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ [1]FREE pop-up blocking with the new MSN Toolbar get it now! References 1. http://g.msn.com/8HMBENUS/2728??PS=47575 From Barry.R.Rittman <@t> uth.tmc.edu Fri May 7 15:13:40 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] tissue slice tool Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0635A5B@UTHEVS3.mail.uthouston.edu> Wendy providing your tissue is not too thick, you can hold two hard backed razor blades together with a screw type tubing clamp. Placing a metal spacer between the backs will give you the thickness of slice that you need. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Wendy England Sent: Fri 5/7/2004 2:20 PM To: histonet@pathology.swmed.edu Cc: Subject: [Histonet] tissue slice tool Hello, We are using razor blades to slice tissue. I am wondering if there is a special tool that has two blades 2-3 mm apart so that the slice can be made by one cut and slice should be more flat. Anybody has idea where I can find it or it's just not practical? Thanks, Wendy _________________________________________________________________ Watch LIVE baseball games on your computer with MLB.TV, included with MSN Premium! http://join.msn.com/?page=features/mlb&pgmarket=en-us/go/onm00200439ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From joseph-galbraith <@t> uiowa.edu Fri May 7 15:22:38 2004 From: joseph-galbraith <@t> uiowa.edu (Galbraith, Joe) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] AFB= bad ctrl for Fite, Negative controls, decloaker, egfr Message-ID: <5D03ED7B9391D4119D9B0008C76B7B2403005B1D@uihc-mail1.uihc.uiowa.edu> RE EGFR: We also found that the Dako EGFR kit gave a higher number of positives than our previous clone H11 (also from Dako). Dako claims that the new and expensive kit has less cross reactivity with other Her molecules when compared to the H11 clone. We have also switched to the pharmDx kit since it is required to make the decision to treat with Erbitux. With some tweaking I would not be surprised to find that the H11 clone could be brought up to a level of positivity similar to the kit but the problem of cross reactivity would still present a problem, hence we acquiesced. Best wishes, Joe Galbraith -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of JOHN COLEMAN Sent: Friday, May 07, 2004 1:48 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] AFB= bad ctrl for Fite, Negative controls, decloaker,egfr Using an AFB control for a Fite stain is a dangerous game! It is quite possible for the control to look positive when the capsule of the bacteria in question on the test slide is present and dissolved and the slide therefore looks negative! For Negative controls, in addition to matching all the rest of the protocol like you already are doing, you must at least also apply the same diluent used for the primary antibodies since letting the slide dry out will in most cases alter the stainability of the slide if there were an issue of false positivity gotten from the secondary reagents, retrieval or other ancillary procedures or reagents. Biocare's decloaker is basically a fine tuned lab grade pressure cooker- we use neither. We tried the pressure cooker offered by Cellmarque, and it saved us no time, and did not significantly improve retrieval.What antibody are you having trouble with? I could tell you what we do if it's one we use. I tested 6 EGFRs on 45 breast cancer and 30 colon cancers looking for a percentage that matched that of Dako's kit and I have to say that the kit's clone is the best I've found. The other clones have a 5-10 % staining of all cases and The Dako kit had a yield closer to 50%, which is what the research for the drug claims. We bit the bullet ( price tag) and now order the kit. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From louri_c <@t> hotmail.com Fri May 7 16:14:12 2004 From: louri_c <@t> hotmail.com (Louri) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Fax Message Received Message-ID: From joseph-galbraith <@t> uiowa.edu Fri May 7 15:11:53 2004 From: joseph-galbraith <@t> uiowa.edu (Galbraith, Joe) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] acetyl-cholinesterase Message-ID: <5D03ED7B9391D4119D9B0008C76B7B24030085FA@uihc-mail1.uihc.uiowa.edu> Peggy: We make our own reagents for ACHE. Our method works pretty well. Email me if you need a protocol. Joe Galbraith -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of DiCarlo, Margaret Sent: Friday, May 07, 2004 12:21 PM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] acetyl-cholinesterase Histonetters, Is anyone out there using acetyl-cholinesterase for aganglionosis? If so where where do you purchase your kit? Thanks for your help. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to 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, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From joseph-galbraith <@t> uiowa.edu Fri May 7 15:33:34 2004 From: joseph-galbraith <@t> uiowa.edu (Galbraith, Joe) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] What's cooking...? Only slides Message-ID: <5D03ED7B9391D4119D9B0008C76B7B2403005B1E@uihc-mail1.uihc.uiowa.edu> MinHan: We get excellent results with a microwave pressure cooker from Nordicware. The time at temperature and pressure is 4 minutes, but it takes 10 minutes to get up to temp/press for a total time in the microwave of 14 minutes. We have a Biocare laboratory 'Decloaker' pressure cooker but even when the company experts come to work with us, our inexpensive microwave version consistently works better (much to the frustration of the company reps). We even get good to excellent retention of morphology in addition to far better antigen retrieval than all other systems we have tested (and we have tried them all). Good luck, Joe Galbraith -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Tan, MinHan Sent: Friday, May 07, 2004 9:52 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] What's cooking...? Only slides Good morning, We are planning to purchase a pressure cooker immediately for a particularly troublesome antigen Unfortunately, no one here has used one before for antigen retrieval, and we have a couple of questions (which may seem rather silly). a. Most posts on Histonet seem to have used either the Nordicware cooker ($45) or the Biocare decloaker ($?). Does a 'decloaker' work differently from a pressure cooker? b. Pressure cooking of slides - most protocols seems to range from 3 min - 5 min. I haven't done much cooking myself (thanks to a wonderful wife), but others in my lab assure me that it takes time for a cooker to build up pressure. In steam retrieval, I usually check to ensure that the buffer is at 95 deg C before starting AR. Is there some kind of equivalent for this in 'pressure cooking'? Thanks! (and apologies for my culinary incompetence) Min-Han Tan 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From degaboh <@t> rice.edu Fri May 7 16:10:16 2004 From: degaboh <@t> rice.edu (ZP) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] fixing and mounting swollen microparticles Message-ID: <20040507211018.E79341DBC6@handler1.mail.rice.edu> Hello all, I have gelatin microparticles that will be loaded with a growth factor by swelling them in the growth factor solution, and I want to see the distribution of the growth factor in the particles with IHC. Ideally, it would be nice if the particles stay swollen, but I am going to be cryosectioning and fixing them in acetone. If I'm going to be fixing in acetone anyway (which will dehydrate them), does it matter if I use xylene/Permount (which dehydrates and thus may shrink the particles) vs. the aqueous mount? Thanks! Zarana Patel degaboh@rice.edu From ploykasek <@t> phenopath.com Fri May 7 18:23:22 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Linear stainer Message-ID: Hi all. I'm looking for a linear stainer for H&E's, but I can't remember the manufacturer. Does anyone know the name? It's the "bicycle chain" type stainer. I used one a long time ago, and for H&E's it was a good workhorse. Thanks for the help. Patti Loykasek From histojock <@t> hotmail.com Fri May 7 19:03:31 2004 From: histojock <@t> hotmail.com (Histo Jock) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Tissue Array Message-ID: You might want to be careful using Zymed's arrays. I don't think they are made with the standard coring method. Rather, I think that they use some sort of melting / re-embedding process that can effect staining down the road. HistoJock -----Original Message----- From: Scott Mordue Subject: Re: [Histonet] Tissue array To: histonet@pathology.swmed.edu Message-ID: <20040430164416.15176.qmail@web42002.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hi Yan Gao, I think you should contact Zymed. They have very high quality Tissue Microarray, especially the human ones. 800-874-4494 Scott CSU -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of yan gao Sent: Thursday, April 29, 2004 7:11 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Tissue array Hi, Histonet. I am interested to get a few tissue array for different human cancers and cell array from human tumor cell lines. Any recommendation? Yan Gao Ph.D Norvatis _________________________________________________________________ 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 georgecole <@t> ev1.net Sat May 8 12:00:38 2004 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] (no subject) Message-ID: <000001c4351d$fde4f1e0$054dbad0@hppav> Histotechs; It has been fun, the memories of ---ahem---shall we say-some of the more experienced Histotechs among us--- those that have a pile of old pages below their calendars, torn off by he rub of time----but this old tech, retired and knee deep in old calendars, has a Wonder on me: I don't see much of a grasp ---in the patter of Cyber Keys---of just how rich a turn over there has been in those pages. June, 1962, freshly thrust into Histology by hooking up with a Neuroresearch lab to do the histo search through whole rat. Cat and human brains by Doctors Robert S. Dow, leading neurologist, and Dr. Olaf Larsell, Grand anatomist ( he was the one that standardized the nomenclature of the cerebellum)---I---freshman histotech took to looking for minute lesions of electrode tracts and reflected lesions of upstream stimuli----doing Neutral Red and Hematoxylin stains and Bodian "Strong Silver Protein" studies on large portions of brains-and some time the whole brains---of study subjects. Downstairs in the hospital proper, the histotechs were ever at their microtomes cutting paraffin sections for a very limited range of stains----timid peons in the shadows of the Giants--- Medical Tecnicians-Tah Dah!! But here I open my eyes every morning on the HIstonet, with hundreds of histotechs snowing the beejeemany out of me with the names of techniques and findings and recipes a good portion of which are communicated with little knots of letters only----and I sort of duck to keep from being blown away by the stupendous riches of technique that are let loose every day in those lines on my HP screen. (Ha!!---look at me---I did it too---HP---2 letters behind which my just now growing experience with computers hides) . Anyway, if you care to know, you do not have to genuflect before the MT's anymore. They have machines with funnels on them into which they pour what they don't especially know much about----but the hands-on life of you Histotechs with immunos, wide, wide studies of everything from ditsy bits of chromosomes to full and complete lay outs of every tissue in captivity----folks-No longer do-you need not to genuflect toward the MT Lab or huddle against the side of the hall as one of Them passes. You are Histoetchs sifting through the many furnishings of Life, hauling out data that even the Mayo Brothers knew naught a wot of those 42 years ago when I first met the animal and Human components coming off my rotary Microtome, Sliding Microtome and Cryostat. Walk head up, you New Ones----and when you meet a Med Tech, head on in the hall,---flick imaginary lint off your sleeve, and with head at a noble angle up,---murmer----"Don't Know Ya, Don't Know Ya-----!" georgecole@ev1.net From oneil <@t> wmca.net Sat May 8 16:10:03 2004 From: oneil <@t> wmca.net (O'Neil) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Help for a Histotech Message-ID: <003901c43540$d5b2a410$a9432a42@sam> I am currently trying to get certified to be a histotechnician and I am looking for tissue including Artery, Liver, Lung, Skin to include epithelium, Kidney to include cortex and medulla, Tonsil, Uterus to include endometrium and myometrium, Small intestine to include all layers, and any AFB tissue. If there is anyone out there who has any information that can help me please e-mail me at oneil@wmca.net, or mail the tissues to Wasatch Histo Consultants Attn: Martena 80 Youngberg Road Winnemucca, NV 89445 I would appreciate any information and/or help anyone can give me. Thank You, Martena Ramos From sonyalhogg <@t> yahoo.co.nz Sat May 8 19:27:56 2004 From: sonyalhogg <@t> yahoo.co.nz (=?iso-8859-1?q?Sonya=20Hogg?=) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] HT practical exam blocks and slides Message-ID: <20040509002756.81665.qmail@web14924.mail.yahoo.com> Hello:) I have read my practical instructions list and they want slides and blocks labelled permanently. How have all of you done this? My lab doesnt have a block labelling machine like some other labs. We just label the blocks and (frosted ended) slides with pencil, then relabel the slides once cut and stained with a sticky label. Any advice would be appreciated???? Thanks Find local movie times and trailers on Yahoo! Movies. http://au.movies.yahoo.com From Rcartun <@t> harthosp.org Sun May 9 09:13:30 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Flow cytometry & IHC billing problems Message-ID: For this of you doing clinical pathology testing, is anyone seeing insurance denials in situations when flow cytometry and immunohistochemistry are performed on the same patient specimen (e.g., lymphoma work-up on a lymph node)? Thank you. Richard Cartun From carl.hobbs <@t> kcl.ac.uk Sat May 8 14:25:51 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] re Nordicware pressure cooker References: <200405081701.i48H1cb0013035@redwood.kcl.ac.uk> Message-ID: <000d01c435f8$e008ef60$40019b51@home> Joe, I too use the Nordicware M/W pcooker; consistent , excellent results for the past 5yrs . Just interested in the solution/s you use to disclose your Ags.....Be grateful for info --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.668 / Virus Database: 430 - Release Date: 24/04/2004 From carl.hobbs <@t> kcl.ac.uk Sun May 9 14:56:02 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] re GCole Message-ID: <002a01c435ff$a86cc610$40019b51@home> Hey...... Well put, Sir. I bet each age had it's technical connundrums/ dilemmas which were processed to the best by the best. Same problems, different technology. No harder, no easier. I remember being berated by Wallington( "Drury and...." the English bible of the time and damn fine reading) for using DPX over Canada balsam...boy! did he blow me out of the water. However....I'm still using DPX. Sorry --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.668 / Virus Database: 430 - Release Date: 24/04/2004 From jhnspam <@t> aol.com Sun May 9 17:28:53 2004 From: jhnspam <@t> aol.com (jhnspam@aol.com) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Cerner Millennium Message-ID: <8c.a6d619a.2dd00aa5@aol.com> I beleive the Shandon microwriter is the only one that will interface. I now for certain that the interface has already been written and is available with rev 8. Pam Johnson From jhnspam <@t> aol.com Sun May 9 17:33:44 2004 From: jhnspam <@t> aol.com (jhnspam@aol.com) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Linear stainer Message-ID: Thermo Electron (Shandon) is the vendor. Where are you from? Pam Johnson From bcfinlay <@t> cox.net Sun May 9 18:35:32 2004 From: bcfinlay <@t> cox.net (bcfinlay@cox.net) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] HT practical exam blocks and slides Message-ID: <20040509233532.EDXS19459.lakermmtao07.cox.net@smtp.east.cox.net> For my HT practical, I used a Secureline marker on the slides and then I hand wrote labels out with a regular pen. We also use pencils for our blocks, but I decided to use the Secureline marker on my blocks as well. I processed my tissue first and then embedded the tissues in different blocks labeled with the marker. The marker won't work on our cassettes through processing and that's the reason I did it this way. From tissuearray <@t> hotmail.com Sun May 9 23:49:52 2004 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Tissue Array Message-ID: How does melting paraffin embedded tissues effect the staining down the road? That doesn't make since. I have made dozens of multiple punch arrays by melting the punches and embedding them as you would normally embed tissues and it has never effected the staining, "DOWN THE ROAD....." Thom for more information on Tissue Microarray Instruction visit: [1]www.arrayworkshop.com >From: "Histo Jock" >To: Histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Tissue Array >Date: Fri, 07 May 2004 20:03:31 -0400 > > >You might want to be careful using Zymed's arrays. I don't think >they are made with the standard coring method. Rather, I think that >they use some sort of melting / re-embedding process that can effect >staining down the road. > >HistoJock > >-----Original Message----- > >From: Scott Mordue >Subject: Re: [Histonet] Tissue array >To: histonet@pathology.swmed.edu >Message-ID: <20040430164416.15176.qmail@web42002.mail.yahoo.com> >Content-Type: text/plain; charset=us-ascii > >Hi Yan Gao, > >I think you should contact Zymed. They have very high >quality Tissue Microarray, especially the human ones. >800-874-4494 > >Scott >CSU > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On >Behalf Of yan gao >Sent: Thursday, April 29, 2004 7:11 PM >To: histonet@pathology.swmed.edu >Subject: [Histonet] Tissue array > > > > Hi, Histonet. > I am interested to get a few tissue array for >different human cancers > and cell array from human tumor cell lines. Any >recommendation? > > Yan Gao > Ph.D > Norvatis > >_________________________________________________________________ >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 _________________________________________________________________ [2]Check out the coupons and bargains on MSN Offers! References 1. http://www.arrayworkshop.com/ 2. http://g.msn.com/8HMBENUS/2743??PS=47575 From Megan.Kear <@t> hunter.health.nsw.gov.au Mon May 10 02:30:05 2004 From: Megan.Kear <@t> hunter.health.nsw.gov.au (Megan Kear) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] EY laboratories information Message-ID: Hi histonetters I am looking for a supplier for the polyclonal antibody to Laminin from EY laboratories, san Mateo, California, USA that distributes to australia. Thank you for your help. M Kear HAPS Australia From M.Bromley <@t> dgri.scot.nhs.uk Mon May 10 03:33:29 2004 From: M.Bromley <@t> dgri.scot.nhs.uk (Mike Bromley) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Cytology textbook Message-ID: <7325D637DFE2D211928800902733A7F303B94AD8@DGAMTBDCEMS> Hi All Do you have any recommendations for a cytology book? Preferably one with both technical procedures and pathology Best Wishes Mike Bromley Chief Biomedical Scientist Pathology Dumfries & Galloway Royal Infirmary Scotland, UK > This e-mail and any files transmitted with it are private and intended > solely for the use of the individual or entity to whom they are addressed. > If you have received this e-mail in error please return it to the address > it > came from telling them it is not for you and then delete it from your > system. > > From juan.gutierrez <@t> christushealth.org Mon May 10 06:36:16 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] acetyl-cholinesterase Message-ID: We make our own. Be glad to send you a copy. Godd luck. Juan C. Gutierrez, HT(ASCP) Histology Supervisor Christus Santa Rosa Hospital 333 N. Santa Rosa Ave. San Antonio, TX 78207 -----Original Message----- From: DiCarlo, Margaret [mailto:MDiCarlo@KaleidaHealth.Org] Sent: Friday, May 07, 2004 12:21 PM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] acetyl-cholinesterase Histonetters, Is anyone out there using acetyl-cholinesterase for aganglionosis? If so where where do you purchase your kit? Thanks for your help. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to 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, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jo-ann <@t> lan1.molonc.mcgill.ca Mon May 10 09:32:14 2004 From: jo-ann <@t> lan1.molonc.mcgill.ca (jo-ann) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Help - Zebra Fish Message-ID: Hi Everyone, My new challenge for this week is that someone has asked me to process Zebra Fish embryos. I haven't got a clue of a processing schedule. Can someone help please. I am meeting with the Investigator tomorrow. Thanks in advance Jo-Ann Bader From KMB1904 <@t> aol.com Mon May 10 09:55:25 2004 From: KMB1904 <@t> aol.com (KMB1904@aol.com) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Kidney and muscle biopsy send out specimens from Delaware Message-ID: <5BEDCCFA.6A1EBF93.00164D58@aol.com> If there is anyone out there that can help us, I would appreciate it...In the past we have sent our muscle biopsies and kidney biopsies along with questionable cases out to the Armed Forced Institute of Pathology. They have recently raised their prices dramatically and we are looking for other options...OUR optimum choice would be somewhere that would bill medicare and medicaid. If anyone has any suggestions, I could use all the help I can get!! Thank you Kathy Bowden Nanticoke memorial Hospital Seaford, De. 19973 From tflore <@t> lsuhsc.edu Mon May 10 10:11:50 2004 From: tflore <@t> lsuhsc.edu (Flores, Teresa) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Kidney and muscle biopsy send out specimens from D elaware Message-ID: Kathy, what is raise prices dramatically mean? Our Diagnostic EM Lab is CAP Inspected and CLIA approved and provides the following: Immunofluorescent renal cases are tagged with IgG, IgA, IgM, C3, C1q, Kappa and Lambda and C4d if it is a transplant kidney. High Resolution Light Microscopy Transmission Electron Microscopy For a fee of $800.00 If STAT studies are required on IF, HRLM, or TEM, there is an additional fee of $200.00 for each technique requested STAT. Teresa Flores LSUHSC Diagnostic EM Lab New Orleans, LA Please view our Web Page http://www.medschool.lsuhsc.edu/pathology/pathist Original Message----- From: KMB1904@aol.com [mailto:KMB1904@aol.com] Sent: Monday, May 10, 2004 9:55 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Kidney and muscle biopsy send out specimens from Delaware If there is anyone out there that can help us, I would appreciate it...In the past we have sent our muscle biopsies and kidney biopsies along with questionable cases out to the Armed Forced Institute of Pathology. They have recently raised their prices dramatically and we are looking for other options...OUR optimum choice would be somewhere that would bill medicare and medicaid. If anyone has any suggestions, I could use all the help I can get!! Thank you Kathy Bowden Nanticoke memorial Hospital Seaford, De. 19973 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon May 10 10:19:27 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] HT practical exam blocks and slides In-Reply-To: <20040509233532.EDXS19459.lakermmtao07.cox.net@smtp.east.co x.net> Message-ID: <3.0.6.32.20040510091927.00c17dd0@gemini.msu.montana.edu> We never lose (during processing) Secureline marking on our cassettes as long as we use the black pen, but if you ever use the red Secureline, you are in HUGE trouble. At 07:35 PM 5/9/2004 -0400, you wrote: >For my HT practical, I used a Secureline marker on the slides and then I hand wrote labels out with a regular pen. We also use pencils for our blocks, but I decided to use the Secureline marker on my blocks as well. > >I processed my tissue first and then embedded the tissues in different blocks labeled with the marker. The marker won't work on our cassettes through processing and that's the reason I did it this way. > > >_______________________________________________ >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 convmcm <@t> cc.usu.edu Mon May 10 10:34:13 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] HT practical exam blocks and slides In-Reply-To: <20040509233532.EDXS19459.lakermmtao07.cox.net@smtp.east.cox.net> Message-ID: <001701c436a4$3f062400$4a737b81@Cygnus> Happy Monday Morning, everyone!!! I used to use Secureline markers, but I found sometimes the writing comes off during deparaffinizing. These markers are great when they work right, but it's not at all great when the writing comes off. I pencil the case number on the slides, then apply a sticker label with printed information after staining is completed. This works well for me and I assume this is acceptable to the ASCP. I'm interested in what the rest of you have to say about this, too. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of bcfinlay@cox.net Sent: Sunday, May 09, 2004 4:36 PM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] HT practical exam blocks and slides For my HT practical, I used a Secureline marker on the slides and then I hand wrote labels out with a regular pen. We also use pencils for our blocks, but I decided to use the Secureline marker on my blocks as well. I processed my tissue first and then embedded the tissues in different blocks labeled with the marker. The marker won't work on our cassettes through processing and that's the reason I did it this way. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From convmcm <@t> cc.usu.edu Mon May 10 10:43:12 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] mystery message from this list In-Reply-To: Message-ID: <001801c436a5$8054a070$4a737b81@Cygnus> HI all. I have received two messages from a sender named "Louri" with the Histonet name in the subj. In one message, there was only an attachment (which I DID NOT open) in the message and in the other message was the instructions to fax the attached file. I'm assuming this is some kind of virus or worm or something. Anyone else get this? Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 From tpmorken <@t> labvision.com Mon May 10 11:03:31 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] mystery message from this list Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA21775D1@usca0082k08.labvision.apogent.com> Connie, the spammers have found histonet - I've also been getting other messages as well marked "histonet" that are spam. Nothing is safe for long on the web. Tim Morken -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Monday, May 10, 2004 8:43 AM To: 'jo-ann'; Histonet@lists.utsouthwestern.edu Subject: [Histonet] mystery message from this list HI all. I have received two messages from a sender named "Louri" with the Histonet name in the subj. In one message, there was only an attachment (which I DID NOT open) in the message and in the other message was the instructions to fax the attached file. I'm assuming this is some kind of virus or worm or something. Anyone else get this? Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From convmcm <@t> cc.usu.edu Mon May 10 11:00:30 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Help - Zebra Fish In-Reply-To: Message-ID: <001901c436a7$eadbd3d0$4a737b81@Cygnus> I'm not an expert in fish histology, but embryos, being very fragile, would be best processed with minimal times. I'm assuming you will be doing these in paraffin. After being properly fixed, I would process them, in ethanol, 70%, 80%, 95%, absolute, absolute + xylene, xylene (2 changes / 15 minutes each), then paraffin, 58 C, 4 changes, 30 minutes each change. I prefer xylenes, not the substitutes, but if that's what you use... I use this protocol for mouse livers and any other small, fragile tissues that come through here and it works wonderfully. Hope this helps Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of jo-ann Sent: Monday, May 10, 2004 7:32 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Help - Zebra Fish Hi Everyone, My new challenge for this week is that someone has asked me to process Zebra Fish embryos. I haven't got a clue of a processing schedule. Can someone help please. I am meeting with the Investigator tomorrow. Thanks in advance Jo-Ann Bader _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Mon May 10 11:04:09 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] mystery message from this list Message-ID: Yes, and made the same assumption. Juan -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Monday, May 10, 2004 10:43 AM To: 'jo-ann'; Histonet@lists.utsouthwestern.edu Subject: [Histonet] mystery message from this list HI all. I have received two messages from a sender named "Louri" with the Histonet name in the subj. In one message, there was only an attachment (which I DID NOT open) in the message and in the other message was the instructions to fax the attached file. I'm assuming this is some kind of virus or worm or something. Anyone else get this? Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mkundu <@t> purdue.edu Mon May 10 11:05:03 2004 From: mkundu <@t> purdue.edu (Moumita Kundu) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] mystery message from this list References: <001801c436a5$8054a070$4a737b81@Cygnus> Message-ID: <000701c436a8$8da939e0$4496d280@agriculture.purdue.edu> yes i got it too.I just deleted . ----- Original Message ----- From: "Connie McManus" To: "'jo-ann'" ; Sent: Monday, May 10, 2004 10:43 AM Subject: [Histonet] mystery message from this list > HI all. > > I have received two messages from a sender named "Louri" with the > Histonet name in the subj. In one message, there was only an attachment > (which I DID NOT open) in the message and in the other message was the > instructions to fax the attached file. I'm assuming this is some kind > of virus or worm or something. Anyone else get this? > > > Connie McManus > Utah Veterinary Diagnostics Laboratory > Utah State University > Logan, UT > Phone: 435/797-1891 > fax: 435/797-2805 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From LINDA.MARGRAF <@t> childrens.com Mon May 10 11:14:12 2004 From: LINDA.MARGRAF <@t> childrens.com (LINDA MARGRAF) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] mystery message from this list Message-ID: Dear Connie and all the Histonetters: I agree this mystery message looks like it must be spam or a virus. I have removed the sender from the list membership. I hope that helps for a while. Everyday I delete 80 or so messages that are trying to post to the list from viruses, advertisers etc. I guess this one got thru. Linda M Histonet administrator From convmcm <@t> cc.usu.edu Mon May 10 11:48:00 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Flow cytometry & IHC billing problems In-Reply-To: Message-ID: <001a01c436ae$8df1f3f0$4a737b81@Cygnus> I don't work in clinical, but I am curious about this. Why would insurance deny this??? Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard Cartun Sent: Sunday, May 09, 2004 7:14 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Flow cytometry & IHC billing problems For this of you doing clinical pathology testing, is anyone seeing insurance denials in situations when flow cytometry and immunohistochemistry are performed on the same patient specimen (e.g., lymphoma work-up on a lymph node)? Thank you. Richard Cartun _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carl.hobbs <@t> kcl.ac.uk Mon May 10 12:19:41 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] re zebrafish Message-ID: <006201c436b2$fb5ea920$43a79951@home> Concur with Connie! I use several techniques...depending upon the requirements. Can be frozen, pwax or GMA resin. I use a similar processing schedule to Connie..., I use IMS, my xylene times are longer. If you want morphology only, can't beat GMA( also works very well for whole-mount ISH- treated embryos). NB yolk-sac can be a problem tho, with pwax IMHO --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.668 / Virus Database: 430 - Release Date: 24/04/2004 From bakerj <@t> umich.edu Mon May 10 12:17:35 2004 From: bakerj <@t> umich.edu (John Baker) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] normal tibia histology in mice Message-ID: Hello- We are running a study looking at C57BL/6j mice and C3H mice which were fed diets of high, normal or low calcium. We were wondering if anyone knows of any articles that did H&E staining on these strains of mice specifically on the tibia? It does not need to be a study in which there were different calcium diets we just want something to compare the results that we are seeing. If you have any thoughts or ideas on where to look it would be greatly appreciated. Please e-mail me at oyserman@umich.edu or John Baker at bakerj@umich.edu. Sincerely, Sivan Oyserman ORL University of Michigan -- ------------------------------------------------------- Today is the Tomorrow you worried about Yesterday. Was it worth it? Birthdays are good for you. Statistics show that the people who have the most live the longest. ------------------------------------------------------- John A. Baker The University of Michigan Orthopaedic Research Laboratories Histology Unit 400 North Ingalls, G160 Ann Arbor, MI 48109-0486 Main lab office phone:734-763-9674 Histology office:734-936-1635 From HACKERLAB <@t> aol.com Mon May 10 11:38:22 2004 From: HACKERLAB <@t> aol.com (HACKERLAB@aol.com) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Linear stainer Message-ID: <196.295f00f6.2dd109fe@aol.com> Dear Patti; I know someone answered back that Shandon is the manufacturer of the Linear Stainer. This is not correct. Hacker Industries Inc has been manufacturing and distributing this product for over 15 years. Please do give us a call if we can be of assistance. Best regards, Elfi Hacker Hacker Industries inc. (803) 712-6100 or 1-800-4-HACKER From HACKERLAB <@t> aol.com Mon May 10 11:58:55 2004 From: HACKERLAB <@t> aol.com (HACKERLAB@aol.com) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Linear Stainer Message-ID: Dear Pam; The Linear Stainer is and always has been manufactured by Hacker Industries Inc. Best regards, Elfi Hacker HACKER Industries Inc (803) 712-6100 or 1-800-4-HACKER Thermo Electron (Shandon) is the vendor. Where are you from? Pam Johnson From Xudong_Cao <@t> brown.edu Mon May 10 13:45:06 2004 From: Xudong_Cao <@t> brown.edu (Cao, Xudong) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] double staining for nerve fibers and endothelial cells on skin samples Message-ID: <1DA88DDC48CCC245A777599FDAEED6D0A38FEE@MAIL1.AD.Brown.Edu> Dear all: I am trying to determine the time and spacial sequence of nerve innervation and angiogenesis of a grafted skin equivalent using histo-chemical-immunostaining. The primary antibodies that I will be using for this study are PGP9.5 for nerve fiber and CD-31 (i.e. PECAM-1) for vascularization, and I will be using two different but compatible peroxidase substrate to visualize the signals . My questions: 1) is there anything that I need to pay special attention to for double staining in general? 2) the staining for nerve fiber normally calls for a thick section (40 to 50 um) whereas that for endothelial cells calls for a relatively thin section (6-8 um). How can I accommodate this difference in thickness, any suggestions? thanks for the help. Xudong Cao, Ph.D. Post-doctoral fellow Brown University From JColCLEFA <@t> aol.com Mon May 10 13:59:32 2004 From: JColCLEFA <@t> aol.com (JColCLEFA@aol.com) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] "disclosure", -secureline- acetyl cholinesterase- musc/kid bx Message-ID: Solutions for antigen "disclosure"- mostly Citra @ 98C for 25min from Biogenex, Trilogy @ 98C for 15 min from Cellmarque (beware of tissue "digestion" from the high surfactant content) and Target retrieval from Dako but only for CD21- all these are dependent on which antibodies you plan to use. we had issues with certain lot numbers of securline pens- usually they are OK but every so often we had a bunch of cassettes or slides lose the writing- now we test the lots before they are put into use- if they don't work we get credit from the vendor. I've had tissues lose antigenicity because of multiple melt downs, but usually after more than three or four times, and probably at temps way too high We do Acetyl cholinesterase stain for aganglionosis, but I make all the reagents myself, I don't use a kit ( yes I'm old school all right- I have a herd of goats, rabbits and mice for my antibodies also-) (kidding)- e mail me directly and I'll send info on reagents, vendors and a no-fail 20 minute procedure-Jcolclefa@aol.com Muscles- we are a reference lab for muscle and renal biopsies,- send me your info and I will forward it to my accounts rep and see what we can do for you. jpcolema@sentara.com as always, questions? e-me at either my work or personal Emails. Call if you like 757-335-2159 From pruegg <@t> colobio.com Mon May 10 14:44:22 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] HT practical exam blocks and slides In-Reply-To: <20040509002756.81665.qmail@web14924.mail.yahoo.com> Message-ID: You could type a slide label and stick it on the slide. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sonya Hogg Sent: Saturday, May 08, 2004 6:28 PM To: Histonet Subject: [Histonet] HT practical exam blocks and slides Hello:) I have read my practical instructions list and they want slides and blocks labelled permanently. How have all of you done this? My lab doesnt have a block labelling machine like some other labs. We just label the blocks and (frosted ended) slides with pencil, then relabel the slides once cut and stained with a sticky label. Any advice would be appreciated???? Thanks Find local movie times and trailers on Yahoo! Movies. http://au.movies.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From convmcm <@t> cc.usu.edu Mon May 10 14:47:39 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] re zebrafish In-Reply-To: <006201c436b2$fb5ea920$43a79951@home> Message-ID: <002601c436c7$a8443290$4a737b81@Cygnus> I agree that plastic is the way to go. I wish with all my heart we were doing plastic -- ANY kind, LRWhite, GMA MMA Whatever --- because it really does make for the best morphology any eye has ever witnessed. Wish wish wish wish... Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Carl Sent: Monday, May 10, 2004 10:20 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] re zebrafish Concur with Connie! I use several techniques...depending upon the requirements. Can be frozen, pwax or GMA resin. I use a similar processing schedule to Connie..., I use IMS, my xylene times are longer. If you want morphology only, can't beat GMA( also works very well for whole-mount ISH- treated embryos). NB yolk-sac can be a problem tho, with pwax IMHO --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.668 / Virus Database: 430 - Release Date: 24/04/2004 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From FreidaC <@t> aol.com Mon May 10 14:27:32 2004 From: FreidaC <@t> aol.com (FreidaC@aol.com) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Linear stainer Message-ID: Shandon does make a linear stainer on which the individual slides are moved through the stains by placing the slide holders on a "bicycle chain". We used one for years and loved it. It was originally an SKI stainer - and was purchased, and is still marketed by Thermo Shandon. They even have a small one that can be used in the frozen section area. Freida Carson From kwittle <@t> jhmi.edu Mon May 10 14:04:33 2004 From: kwittle <@t> jhmi.edu (Karen Wittler) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Tissue slice tool Message-ID: Sakura makes "grossing forks". I have heard them described as a pair of forks in three sizes:1&1/2 mm, 2 mm and 2&1/2mm. They work by inserting the double forks into the tissue securing it while the grosser cuts with a blade on the outside of each fork. Karen Wittler, HT(ASCP) Johns Hopkins Hospital Baltimore, MD From mary.bliss <@t> northwestpathology.com Mon May 10 15:18:43 2004 From: mary.bliss <@t> northwestpathology.com (Bliss, Mary E.) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Breast processing Message-ID: <27066863AD02214B81FE49477F3E71AD0D6F2C42@hinet1.hinet.org> Hi All, How do other laboratorians prepare fatty breast specimens for histology? Are you doing anything special? We had a case today which our doctor needs to have re-processed. It appears unfixed, although it sat in formalin on the processor over the weekend before it processed on Sunday night. The sections are large (too large in my opinion) and not adequately removed of fat. We are using toluene on our processors and have considered going to Xylene. We have tried Penn fixx in the past, but discontinued using it years ago. I know it is a complicated subject, but just thought I would see if anyone has any bits of wisdom. Mary E. Bliss Lead Histologist Northwest Pathology, P.S. 3614 Meridian St. Suite 100 Bellingham, WA 98225 (360)734-2800 x601 (360)734-3818 FAX From pstefanova <@t> sten.sunnybrook.utoronto.ca Mon May 10 15:30:10 2004 From: pstefanova <@t> sten.sunnybrook.utoronto.ca (Petia P Stefanova) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] vibratome References: <002601c436c7$a8443290$4a737b81@Cygnus> Message-ID: <00c301c436cd$977a3260$9d194c8e@WS21203> Hi, I am trying to cut eyes on vibtarome 100 - 150 microns and I find it almost impossible. Does anybody have experience in it? I would appreciate some tips. Thanks Petia From Carolyn.Earley <@t> leica-microsystems.com Mon May 10 15:33:08 2004 From: Carolyn.Earley <@t> leica-microsystems.com (Carolyn.Earley@leica-microsystems.com) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Linear stainer Message-ID: Carolyn Earley To: Patti Loykasek 05/10/2004 01:30 cc: histonet-bounces@lists.utsouthwestern.edu PM Subject: Re: [Histonet] Linear stainer(Document link: Carolyn Earley) Hi Patti, There are several Linear Stainers on the market currently. Hacker and Leica Microsystems both make a batch type linear stainer that is capable of staining 20-30 slides per rack in a linear fashion. I do believe, however, that the bicycle-chain type stainer you are inquiring about is only made by Thermo-Shandon. Realize, however, that this stainer uses individual clips to hold the slides which are loaded continuously one at a time. It is a workhorse, as you say. Good luck in your search for a stainer and to make your search even easier, click onto the web sites of the companies mentioned and have a look at what we have to offer. Carolyn Earley Marketing Manager Leica Microsystems 847-405-7014 Carolyn.Earley@Leica-Microsystems.com Patti Loykasek To: histonet Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] Linear stainer western.edu 05/07/2004 06:23 PM Hi all. I'm looking for a linear stainer for H&E's, but I can't remember the manufacturer. Does anyone know the name? It's the "bicycle chain" type stainer. I used one a long time ago, and for H&E's it was a good workhorse. Thanks for the help. Patti Loykasek _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From convmcm <@t> cc.usu.edu Mon May 10 15:52:19 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] mystery message from this list In-Reply-To: <000701c436a8$8da939e0$4496d280@agriculture.purdue.edu> Message-ID: <003301c436d0$af9854f0$4a737b81@Cygnus> Thanks for letting me know you have all seen this same thing. Linda Margraf, thank you so much for deleting this person from the list. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Moumita Kundu Sent: Monday, May 10, 2004 9:05 AM To: Connie McManus; 'jo-ann'; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] mystery message from this list yes i got it too.I just deleted . ----- Original Message ----- From: "Connie McManus" To: "'jo-ann'" ; Sent: Monday, May 10, 2004 10:43 AM Subject: [Histonet] mystery message from this list > HI all. > > I have received two messages from a sender named "Louri" with the > Histonet name in the subj. In one message, there was only an attachment > (which I DID NOT open) in the message and in the other message was the > instructions to fax the attached file. I'm assuming this is some kind > of virus or worm or something. Anyone else get this? > > > Connie McManus > Utah Veterinary Diagnostics Laboratory > Utah State University > Logan, UT > Phone: 435/797-1891 > fax: 435/797-2805 > > > > _______________________________________________ > 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 convmcm <@t> cc.usu.edu Mon May 10 16:31:07 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Breast processing In-Reply-To: <27066863AD02214B81FE49477F3E71AD0D6F2C42@hinet1.hinet.org> Message-ID: <003901c436d6$1b250a10$4a737b81@Cygnus> Mary, my heart goes out to you. I believe every histotech in the world has gone through this same nightmare --- tissues that are too big and not properly processed, and the pathologist expects YOU, the histotech, to produce perfect results. >From your story, fixation may not be the only problem. 48 hours is generally plenty of time for tissues to fix. But if there is a lot of fatty tissue and not enough room for the fixative to work through the tissue (as in a tissue smushed up inside a processing cassette), you may have a fixation problem compounded with a processing problem. The only solution is for the person trimming in the tissues to keep them between 2 and 3 mm in thickness and to try to keep the length and width dimensions to something less than that of the cassette. There should absolutely not be any tissue sticking out of the cassette nor should there be imprints of the cassette on the tissue. As for the fat in your tissues, if there was adequate processing, it would be gone. Alcohols, toluene, xylene and xyl substitutes do a good job of that. There's nothing quite like good reagent flow in and around the tissue while processing ... Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bliss, Mary E. Sent: Monday, May 10, 2004 1:19 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Breast processing Hi All, How do other laboratorians prepare fatty breast specimens for histology? Are you doing anything special? We had a case today which our doctor needs to have re-processed. It appears unfixed, although it sat in formalin on the processor over the weekend before it processed on Sunday night. The sections are large (too large in my opinion) and not adequately removed of fat. We are using toluene on our processors and have considered going to Xylene. We have tried Penn fixx in the past, but discontinued using it years ago. I know it is a complicated subject, but just thought I would see if anyone has any bits of wisdom. Mary E. Bliss Lead Histologist Northwest Pathology, P.S. 3614 Meridian St. Suite 100 Bellingham, WA 98225 (360)734-2800 x601 (360)734-3818 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SJones <@t> cvm.tamu.edu Mon May 10 16:59:05 2004 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Breast processing Message-ID: Hi Mary, In the past, I have used an alcohol based fixative to help with this problem. Davidson's fixative comes to mind or Anatech's buffered CBA (I think it is called). I agree with Connie that nothing beats proper trimming with these specimens. The old saying goes (cleaned up a bit) Trash In, Trash Out! Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Bliss, Mary E." 5/10/2004 3:18:43 PM >>> Hi All, How do other laboratorians prepare fatty breast specimens for histology? Are you doing anything special? We had a case today which our doctor needs to have re-processed. It appears unfixed, although it sat in formalin on the processor over the weekend before it processed on Sunday night. The sections are large (too large in my opinion) and not adequately removed of fat. We are using toluene on our processors and have considered going to Xylene. We have tried Penn fixx in the past, but discontinued using it years ago. I know it is a complicated subject, but just thought I would see if anyone has any bits of wisdom. Mary E. Bliss Lead Histologist Northwest Pathology, P.S. 3614 Meridian St. Suite 100 Bellingham, WA 98225 (360)734-2800 x601 (360)734-3818 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BetStark <@t> aol.com Mon May 10 17:06:09 2004 From: BetStark <@t> aol.com (BetStark@aol.com) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] ubsubscribe Message-ID: From WilsonL <@t> jwci.org Mon May 10 19:02:40 2004 From: WilsonL <@t> jwci.org (Lori Wilson) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] ubsubscribe Message-ID: <1CD765A8D6B6DB4F887110A8EEE09CE502BB65DB@smexsvr1.jwci.org> Lori L. Wilson, MD From brucea <@t> unimelb.edu.au Mon May 10 20:59:16 2004 From: brucea <@t> unimelb.edu.au (brucea@unimelb.edu.au) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Dilemma..... Message-ID: Hi everyone I posted a question on the histonet a week ago but I don't think I explained my dilemma clearly enough. I am using alkaline phosphatase histochemistry (not immunohistochemistry!) on PFA-fixed whole marsupial blastocysts and embryos to detect primordial germ cells (PGCs). I use 1mg/ml Fast Blue salt and 1mg/ml Naphthol AS phosphate sodium salt in 0.2M Tris buffer, pH 9.4 for the histochemistry, a standard method for the detection of PGCs. The staining is good but I have been told that it is not permanent. Ideally, after staining these embryos histochemically for PGCs, I would like to dehydrate, paraffin- embed, section and then stain them immunohistochemically (ie, with antibodies) for other germ cell markers and several growth factors. Does anyone know of a histochemical method for the detection of germ cells that would later permit me to do this? We have well-established methods for immunohistochemistry for several germ cell markers in our lab. Thank you to the people who took the time to help last time, Dr Danielle Hickford Research Fellow Department of Zoology, University of Melbourne, Australia 3010. hickford@unimelb.edu.au -- BRUCE ABALOZ PH:61383446282 HISTOLOGIST FAX:61383447909 DEPT.of ZOOLOGY EMAIL: brucea@unimelb.edu.au THE UNIVERSITY Of MELBOURNE. VICTORIA.AUSTRALIA 3010 Nobody can make you feel inferior without your permission. - Eleanor Roosevelt DANCE LIKE NO-ONE'S WATCHING ******************************************************************************** This electronic message and all contents contain information which may be privileged, confidential or otherwise protected from disclosure.The information is intended to be for the addressee only. If you are not the addressee, any disclosure, copy, distribution or use of the contents of this message is prohibited. If you have received this electronic message in error, please notify us immediately and destroy the original message and all copies. From rocan <@t> mac.com Tue May 11 00:50:28 2004 From: rocan <@t> mac.com (rocan@mac.com) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] double staining for nerve fibers and endothelial cells on skin samples In-Reply-To: <1DA88DDC48CCC245A777599FDAEED6D0A38FEE@MAIL1.AD.Brown.Edu> References: <1DA88DDC48CCC245A777599FDAEED6D0A38FEE@MAIL1.AD.Brown.Edu> Message-ID: <1B85CD44-A30F-11D8-8966-000A9589219E@mac.com> Cao, Au Contraire. The CD31 (MEC 13.3 rat anti-mouse), staining is much, much better the thicker your sections are. Check out the publications from D.M. McDonald from UCSF, he has the absolute best microvessel stainings ever published. If you read the fine print of some of his more recent publications you will find two very important clues: 1) thick sections 40 to 100 um; 2) Long incubations at room temperature. It seems to me you are in luck! Go ahead with your thick sections, you will see blood vessel histochem like you've never seen before. It works like a charm. Let me know how it goes, Rocio On May 10, 2004, at 11:45 AM, Cao, Xudong wrote: > Dear all: > > I am trying to determine the time and spacial sequence of nerve > innervation and angiogenesis of a grafted skin equivalent using > histo-chemical-immunostaining. The primary antibodies that I will be > using for this study are PGP9.5 for nerve fiber and CD-31 (i.e. > PECAM-1) for vascularization, and I will be using two different but > compatible peroxidase substrate to visualize the signals . My > questions: 1) is there anything that I need to pay special attention > to for double staining in general? 2) the staining for nerve fiber > normally calls for a thick section (40 to 50 um) whereas that for > endothelial cells calls for a relatively thin section (6-8 um). How > can I accommodate this difference in thickness, any suggestions? > thanks for the help. > > Xudong Cao, Ph.D. > Post-doctoral fellow > Brown University > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sonyalhogg <@t> yahoo.co.nz Tue May 11 01:52:18 2004 From: sonyalhogg <@t> yahoo.co.nz (=?iso-8859-1?q?Sonya=20Hogg?=) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] A big thankyou Message-ID: <20040511065218.47126.qmail@web14929.mail.yahoo.com> To everyone who helped me with the info about labelling of my blocks and slides for the HT practical a BIG THANKS. Find local movie times and trailers on Yahoo! Movies. http://au.movies.yahoo.com From ian.montgomery <@t> bio.gla.ac.uk Tue May 11 04:02:36 2004 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:22:56 2005 Subject: Fw: Re: [Histonet] Re: Polyester wax (with earlier citations) Message-ID: <6.0.1.1.2.20040511095617.0332b400@udcf.gla.ac.uk> Fran, Retiring shortly, 18 May 2007, when you're old, worn out, been there, done that, hand sharpened knives, used Cambridge Rocker's, still think Zenker and Susa are terrific, three years is shortly. Ian. >Reply-To: "Ian Montgomery" >From: "Ian Montgomery" >To: "Ian Montgomery \(Work\)" >Subject: Fw: Re: [Histonet] Re: Polyester wax (with earlier citations) >Date: Mon, 10 May 2004 23:36:40 +0100 >X-Mailer: Microsoft Outlook Express 6.00.2800.1409 >X-GLA-Spam-Scan: R >X-GLA-Spam-Score: 0.0 (/) >X-GLA-Spam-Report: > > >----- Original Message ----- >From: "Fran Lemons" >To: >Sent: Monday, May 10, 2004 3:11 PM >Subject: Fwd: Re: [Histonet] Re: Polyester wax (with earlier citations) > > >Ian, >You should consider writing standard operating procedures before you head >out to pasture...they are required here in the states. > > >>> Ian Montgomery 05/06/04 06:54AM >>> > Here I am in sunny Glasgow living in a time warp. Automation, >what's that? Hand process, make my own stains, sharpen knives, name it and >I'm still doing it. Well apart from wooden blocks, gave that up 30 odd >years ago. I'm such a sad case that if you ask me what a particular >chemical in my store is for I'll tell you exactly and how much is used. Sad >thing is, I'll be retiring shortly and all the knowledge will go with me. >Ian. > > > > >Hi Paul, > > > >Lots of good times; > >but you forgot about the blistered finger tips from sticking those wax > >blocks on wooden blocks(the smaller the block-the bigger the blister!), and > >all the hand processing, which mainly involved much frantic dashing about > >twirling multiple pots of tissue to provide agitation! > >Who can forget about mouth pipetting whilst eating sandwiches at the > >staining bench and brewing the tea over a bunsen during waiting times! > >If I recall correctly, the pipes were full of either St. Bruno flake or > >'Herbal' tobacco and between chuffs of smoke, most of the conversation was > >punctuated by phrases such as "eh up", "y'whot", "gerroff" and "owd ya > >tight", interspersed with "cool man", "crazy daddy" and "all gone". Ahhh... > >sweet memories. > > > >Bryan Hewlett > >Hamilton, Ont. > >Canada > >(formerly Nottingham, England) > > > >----- Original Message ----- > >From: "Paul Bradbury" > >To: ; "HistoNet Server" > > > >Sent: Wednesday, May 05, 2004 3:32 PM > >Subject: Re: [Histonet] Re: Polyester wax (with earlier citations) > > > > > > > Hi Steve, > > > > > > Your descriptions bring back a flood of memories ... the wax tea pot, > > > Leukhart's embedding rings, sticking the blocks onto wooden blocks, > > > taking them off again at the end of the day, etc. Safety precautions had > > > not even been invented in those days. When I first started in Histology, > > > there were five of us in the lab and every single one smoked a pipe. So, > > > embedding, trimming and sticking on the blocks involved three of us ... > > > all chuffing out smoke. The conversations that took place during these > > > times were priceless. Sadly, this opportunity was lost with the advent > > > of automated embedding centres.. > > > > > > All solvents went down the drain, old specimens were dumped into the > > > sink to allow the formalin to drain away. There was no fume hood, so the > > > formalin fumes were thick enough to cut with a knife. In retrospect, > > > dumping solvents and fixatives down the drain was not the best idea!... > > > but at the time, that was standard practice. However, despite these > > > horrendous practices, we are all still alive and well, and all went on > > > to accept senior positions around the world.. > > > > > > There were no productivity units to count, no QC/QA demands (apart from > > > self-imposed ones), no intrusions from mis-guided administrators. We had > > > time to work on our own projects, investigate new procedures, and read > > > journals looking for new methods. We made all of our own reagents from > > > scratch (hematoxylin, Schiff's reagent, fixatives, etc) We sharpened our > > > own knives. The camaraderie was wonderful, there was no bitching or > > > whining, going for a beer at lunchtime was a routine practice. We did a > > > great job, we went home happy, and provided great service > > > > > > I would not give up the new developments in Histology > > > (immunohistochemistry, monoclonal antibodies, disposable knives, or > > > automated stainers, etc ) they have produced quantum leaps in quality > > > and diagnostic accuracy, but I sometimes I despair that the new > > > generation of technologists have missed out on an invaluable learning > > > experience. > > > > > > I firmly believe that I am a better histologist from my experience ... > > > if something didn't work there was no service rep to call for advice, it > > > was up to us to figure it out. The most respected "mentors" on the > > > Histonet (who I won't name to avoid embarrasing them, but you know who I > > > mean) are all the product of this bygone age of Histology. The Histonet > > > serves a great purpose as a knowledge base and resource for advice, but > > > there is no substitute for self-motivated learning. The books and > > > journals are all out there ... waiting. > > > > > > Okay, I am done. Time to get down off my soapbox. The sermon is ended, > > > > > > Paul Bradbury > > > Kamloops, BC > > > Canada > > > (formerly Nottingham, England) > > > > > > > > > > > > > > > > > > > > > > > > > > > _______________________________________________ > > > 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 > >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 ctsblack <@t> capeheart.uct.ac.za Tue May 11 03:56:49 2004 From: ctsblack <@t> capeheart.uct.ac.za (Melanie Black) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Research Diagnstics CD 66 Message-ID: Hi There I am running an antibody from Research Diagnostics - CD 66 (Granulocyte Marker).Has anybody had success with it on formalin fixed tissue, and is so, what antigen retrieval have you found to be successful. I have had no problem with it on unfixed blood smears, BUT no luck on fixed tissue. My antibody is RDI-CD66B-G10F5, and is recommended for formalin fixed tissue, with citrate pre treatment. It also requires an IgM secondary, which I have used!I have tries citrate buffer as well as trypsin. Any suggestions??? Many Thanks Melanie Black. -- 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 juan.gutierrez <@t> christushealth.org Tue May 11 06:28:26 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] mystery message from this list Message-ID: Thank you, and keep up the good work. Your batting average is a heck of a lot better than our administrators. Juan C. Gutierrez, HT(ASCP) Histology Supervisor Christus Santa Rosa Hospital 333 N. Santa Rosa Ave. San Antonio, TX 78207 -----Original Message----- From: LINDA MARGRAF [mailto:LINDA.MARGRAF@childrens.com] Sent: Monday, May 10, 2004 11:14 AM To: convmcm@cc.usu.edu; jo-ann@lan1.molonc.mcgill.ca; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] mystery message from this list Dear Connie and all the Histonetters: I agree this mystery message looks like it must be spam or a virus. I have removed the sender from the list membership. I hope that helps for a while. Everyday I delete 80 or so messages that are trying to post to the list from viruses, advertisers etc. I guess this one got thru. Linda M Histonet administrator _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From browning <@t> HHSC.CA Tue May 11 06:48:48 2004 From: browning <@t> HHSC.CA (Browning Deb) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Breast processing Message-ID: <3AADFB88753AD31189C100902786B91C0E278186@hch_nt_exchange.hhsc.ca> We have seen a significant improvement with all our larger specimens, including fatty breast specimens, since we not only made fixation for 24 hours (minimum) mandatory, but we set up one of our processors (xylene, not toluene) with a longer processing schedule. Not only are our tissues now fixed, but they are now processed as well. There are only 2 difficult things about this: first, convincing the pathologist that the delay is worth the wait; and second, there seems to be a trend that the longer the tissue has to fix and process, this must mean that the pieces in the cassettes can be proportionately larger, right? -----Original Message----- From: Bliss, Mary E. [mailto:mary.bliss@northwestpathology.com] Sent: Monday, May 10, 2004 4:19 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Breast processing Hi All, How do other laboratorians prepare fatty breast specimens for histology? Are you doing anything special? We had a case today which our doctor needs to have re-processed. It appears unfixed, although it sat in formalin on the processor over the weekend before it processed on Sunday night. The sections are large (too large in my opinion) and not adequately removed of fat. We are using toluene on our processors and have considered going to Xylene. We have tried Penn fixx in the past, but discontinued using it years ago. I know it is a complicated subject, but just thought I would see if anyone has any bits of wisdom. Mary E. Bliss Lead Histologist Northwest Pathology, P.S. 3614 Meridian St. Suite 100 Bellingham, WA 98225 (360)734-2800 x601 (360)734-3818 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From REEVEML <@t> shands.ufl.edu Tue May 11 07:11:14 2004 From: REEVEML <@t> shands.ufl.edu (Mary Reeves) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Alternative's to the Steiner Message-ID: I have been given alternatives's to stain Helicobacter pylori, however, what about the other organisms stained by the Steiner method? -spirochetes -legionella -donavan bodies -cat scratch Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 From jlambrey <@t> hotmail.com Tue May 11 07:39:43 2004 From: jlambrey <@t> hotmail.com (Julien Lambrey de Souza) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] crustacean haemolymph Message-ID: Hello all histonetters, A post-doc working in a the next door lab is having a few problems with his haemolymph smears. He is stuck with a cristalising matrix of haemolymph through wich it is difficult to see the cells. Would anybody have a clue to what may be the cause? Here is his message: I'm preparing slides from crustacean haemolymph (isopods, amphipods and decapod shrimp). I've been advised that touching a drop of haemolymph to a slide, followed by simple air-drying, is a method that should work fine. However, following this method, the haemocytes appear to be embedded within a kind of crystalline matrix. This matrix varies in thickness and in the degree to which it obscures the cells, but it is always a problem, and remains so through fixing, staining and associated rinses. If the haemolymph is not smeared, the crystals appear very thick, and cells are suspended within the matrix. Any ideas on what the problem may be with crustacean haemolymph? I'm about to attempt centrifugation of haemolymph + TA buffer, followed by a TA wash and resuspension, but failing that, I'm a bit stuck, so any ideas or suggestions of things to try are very welcome. The basic steps: Slide prep; drop of haemolymph touched to a slide, then smeared (or not) 24h fixation in MFA (85:10:5 methanol, formalin, acetic acid) 10 min tapwater rinse hydrolysis in 5.0 N HCl dip in 0.1 N HCl 120 min stain in Schiff's reagent 3 x 5 min bisulphite rinses 10 min tapwater rinse 3 x 2 min dH2O rinses air dry, store in dark There it is. So if anybody has any suggestions, I will be happy to follow them through to him. Thanks, Julien. _________________________________________________________________ Open your e-mail without having to worry about viruses [1]With MSN Premium Get 2 Months FREE* References 1. http://g.msn.com/8HMBENCA/2737??PS=47575 From histosci <@t> shentel.net Tue May 11 07:42:58 2004 From: histosci <@t> shentel.net (HSRL) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Leica RM2135 microtome/Hacker 3655 Coverslipper for sale Message-ID: <000001c43755$8244a0a0$0200a8c0@HSRLMAIN> Dear Histonetters, We have a Leica RM2135 microtome for sale. It is in great shape and was used daily until a couple of weeks ago. We bought brand new microtomes to replace our used ones. Also, we have a Hacker 3655 Coverslipper that needs some work or would be great for parts. Please note: we are a research laboratory, not a Leica or Hacker sales representative. If you are interested in either of these items, please make us an offer. Have a great day, Sincerely, Beth Poole HSRL, Inc.- A GLP Compliant Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 beth@hsrl.org www.hsrl.org From nperson211 <@t> comcast.net Tue May 11 08:48:50 2004 From: nperson211 <@t> comcast.net (nperson211@comcast.net) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] VEGF in rat Message-ID: <051120041348.10654.40A0D9C20005BDF50000299E2200748184FFCECECD91908C8D9A8F@comcast.net> Good Morning, Histonetters, I am hoping for some suggestions. I have just run VEGF(Santa Cruz mab sc-7269) on FFPE rat brains. I pretreated with RTU proteinase K from Biogenex, and diluted my antibody to 1:300 and incubated for 45 mins. at room temp. In previous runs with the same conditions, I have gotten beautiful staining on a human hemangiosarcoma specimen. In these rat brains, which are controls for an epilepsy study, EVERY neuron has stained positive for VEGF. There is no background, and the stain is cytoplasmic in EVERY neuron. Does anyone have any experience with this phenomenon? I know there has been much discussion about VEGF antibodies and everyone seemed to agree that the Santa Cruz monoclonal was the best, which is my experience as well. Thanks for any thoughts. Nancy Lemke Hermelin Brain Tumor Center Henry Ford Hospital Detroit From GauchV <@t> mail.amc.edu Tue May 11 08:54:39 2004 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Techs on Renal Biopsies Message-ID: I was just wondering how you all handle renal biopsy cases. Do your histotechs go on the biopsies to identify glomeruli, adequacy of tissue,and divide tissue for routine histology, immunofluorescence and electron microscopy studies, etc. or, if not the techs, who handles that in your institution? Thanks in advance for your help, Vicki Gauch AMCH Albany, NY From cathy <@t> wasatchhisto.com Tue May 11 09:07:31 2004 From: cathy <@t> wasatchhisto.com (Cathy Mayton) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] marking blocks and slides Message-ID: <002401c43761$53d4b1e0$a16b3442@selfuggnbwd3wt> Good morning, We use Sharpie brand industrial fine point marker to write on our blocks and slides. The marker goes through the processing and staining with no problems. These markers can be bought at office supply stores or maybe even at your local Wal-Mart, KMart etc. and are less expensive. Hope this helps. Cathy ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ "Quality Histology with a Personal Touch" A GLP Compliant Laboratory Cathy A. Mayton Wasatch Histo Consultants, Inc. 775-625-4425 www.wasatchhisto.com ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ From la.sebree <@t> hosp.wisc.edu Tue May 11 09:30:35 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Techs on Renal Biopsies Message-ID: Hi Vicki, We have a renal lab staffed by a histologist that only does this type of work. She uses hisotechs from the surgical pathology lab as back-ups when she's busy or absent. They (surg. path. techs) train extensively with the renal histotech to be able to function as back-ups. 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 -----Original Message----- From: Vicki Gauch [mailto:GauchV@mail.amc.edu] Sent: Tuesday, May 11, 2004 8:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Techs on Renal Biopsies I was just wondering how you all handle renal biopsy cases. Do your histotechs go on the biopsies to identify glomeruli, adequacy of tissue,and divide tissue for routine histology, immunofluorescence and electron microscopy studies, etc. or, if not the techs, who handles that in your institution? 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 From Myri37 <@t> aol.com Tue May 11 09:34:04 2004 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Mac Neals Ttetrachrome Message-ID: <451BA304.6EB16A9E.0005167B@aol.com> Hi everyone Could someone give me SIGMA Aldrich references for different stains used for MacNeals tetrachrome, because i found different CI for them methylene blue azure A eosinate (is it A2918 ?) methylene violet glycerin (G 2289) thank you very much sincerly Myriam baali natural implant France From kk320 <@t> cam.ac.uk Tue May 11 09:50:55 2004 From: kk320 <@t> cam.ac.uk (K. Knezevic) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Background lacZ staining Message-ID: Hi, I've just lacZ stained some frozen sections of adult mouse tissues (kidney, spleen and lung) and i've got lacZ background on both my transgenic and wildtype tissues as well as (it looks like) specific staining on my transgenic samples. I was wondering if anybody has a protocol or suggestion for eliminating lacZ background. Thanks Kathy From lizchlipala <@t> premierhistology.com Tue May 11 10:33:03 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] ED-A fibronectin antibody in rat Message-ID: <000001c4376d$4369d9f0$74d48a80@LIZ> Hello I need to perform immunohistochemistry for ED-A fibronectin on rat kidneys. I have done a bit of searching on the internet and have only been able to locate one source for this antibody. Its from Abcam (ab6328) from what I get off their web page is that this antibody will work on frozen sections, but there is no information available on formalin fixed paraffin embedded tissues. Does anyone know if this antibody will work on FFPE tissues and is there another source for this antibody other than Abcam that will work in rat? Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP) 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 MCBD, Room A3B40 Boulder, Colorado 80309 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From jcline <@t> wchsys.org Tue May 11 10:39:27 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] marking blocks and slides, Cathy Mayton In-Reply-To: <002401c43761$53d4b1e0$a16b3442@selfuggnbwd3wt> Message-ID: <000601c4376e$248a2520$1d2a14ac@wchsys.org> I have tried this and the writing was so light we had trouble seeing the numbers. How do you keep the writing dark? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Cathy Mayton Sent: Tuesday, May 11, 2004 10:08 AM To: Histonet Subject: [Histonet] marking blocks and slides Good morning, We use Sharpie brand industrial fine point marker to write on our blocks and slides. The marker goes through the processing and staining with no problems. These markers can be bought at office supply stores or maybe even at your local Wal-Mart, KMart etc. and are less expensive. Hope this helps. Cathy ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ "Quality Histology with a Personal Touch" A GLP Compliant Laboratory Cathy A. Mayton Wasatch Histo Consultants, Inc. 775-625-4425 www.wasatchhisto.com ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***** 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 jcline <@t> wchsys.org Tue May 11 10:49:14 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Breast processing, Mary Bliss In-Reply-To: <27066863AD02214B81FE49477F3E71AD0D6F2C42@hinet1.hinet.org> Message-ID: <000701c4376f$829aff30$1d2a14ac@wchsys.org> I have started microwave fixation for my breast cassettes. Of course I have a Hacker TT microwave processor. This works beautifully. I process for one hour at 40 degrees and can process overnight the same day the specimen is grossed. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bliss, Mary E. Sent: Monday, May 10, 2004 4:19 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Breast processing Hi All, How do other laboratorians prepare fatty breast specimens for histology? Are you doing anything special? We had a case today which our doctor needs to have re-processed. It appears unfixed, although it sat in formalin on the processor over the weekend before it processed on Sunday night. The sections are large (too large in my opinion) and not adequately removed of fat. We are using toluene on our processors and have considered going to Xylene. We have tried Penn fixx in the past, but discontinued using it years ago. I know it is a complicated subject, but just thought I would see if anyone has any bits of wisdom. Mary E. Bliss Lead Histologist Northwest Pathology, P.S. 3614 Meridian St. Suite 100 Bellingham, WA 98225 (360)734-2800 x601 (360)734-3818 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***** 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 mkundu <@t> purdue.edu Tue May 11 10:56:04 2004 From: mkundu <@t> purdue.edu (Moumita Kundu) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Cox -2 IHC References: <000001c4376d$4369d9f0$74d48a80@LIZ> Message-ID: <000b01c43770$77251400$4496d280@agriculture.purdue.edu> I am trying to do IHC with Cox -2 ,The protocol I have is a long overnight incubation with TSA kit. I am trying to reduce the time of incubation using Dako's LSAB+ system or LSAB2 . Please pass me any protocol which can be helpful. Thank you in advance. Moumita Kundu ----- Original Message ----- From: "Elizabeth Chlipala" To: "'Histonet'" Sent: Tuesday, May 11, 2004 10:33 AM Subject: [Histonet] ED-A fibronectin antibody in rat > Hello > > I need to perform immunohistochemistry for ED-A fibronectin on rat > kidneys. I have done a bit of searching on the internet and have only > been able to locate one source for this antibody. Its from Abcam > (ab6328) from what I get off their web page is that this antibody will > work on frozen sections, but there is no information available on > formalin fixed paraffin embedded tissues. Does anyone know if this > antibody will work on FFPE tissues and is there another source for this > antibody other than Abcam that will work in rat? Thanks in advance. > > Liz > > Elizabeth A. Chlipala, BS, HTL(ASCP) > 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 > MCBD, Room A3B40 > Boulder, Colorado 80309 > > _________________________________ > This email and any files transmitted with it may contain PRIVILEGED or > CONFIDENTIAL information and may be read or used only by the intended > recipient. If you are not the intended recipient of the email or any of > its attachments, please be advised that you have received this email in > error and that any use, dissemination, distribution, forwarding, > printing, or copying of this email or any attached files is strictly > prohibited. If you have received this email in error, please immediately > purge it and all attachments and notify the sender by reply email or > contact the sender at the number listed above if one is provided. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Rcartun <@t> harthosp.org Tue May 11 10:47:31 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Techs on Renal Biopsies Message-ID: I perform tissue evaluation during kidney biopsies and when I am not available one of my MD colleagues backs me up. We treat these as intraoperative consultations and bill for them. Decisions regarding obtaining another core of tissue are made based on our interpretation. Personally, I feel that it is too much responsibility to place upon a Histotech. My Immunopathology staff (mostly Histotechs with QIHC) will participate in tissue triage (i.e., processing the tissue for EM, IF, and light microscopy) once I have evaluated it. Richard Cartun, Ph.D. Director, Immunopathology Hartford Hospital Hartford, CT 06102 >>> "Vicki Gauch" 05/11/04 09:54AM >>> I was just wondering how you all handle renal biopsy cases. Do your histotechs go on the biopsies to identify glomeruli, adequacy of tissue,and divide tissue for routine histology, immunofluorescence and electron microscopy studies, etc. or, if not the techs, who handles that in your institution? 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 From mbecker <@t> pathlabinc.com Tue May 11 11:07:15 2004 From: mbecker <@t> pathlabinc.com (Michelle Becker) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] marking blocks and slides, Cathy Mayton In-Reply-To: <000601c4376e$248a2520$1d2a14ac@wchsys.org> Message-ID: Have you tried Statmark Pens from StatLab Medical Products? We have had very good luck with them after trying several pens. It writes dark and stays on through both staining and processing procedures. They also last much longer than secureline markers. michb -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joyce Cline Sent: Tuesday, May 11, 2004 10:39 AM To: Histonet Subject: RE: [Histonet] marking blocks and slides, Cathy Mayton I have tried this and the writing was so light we had trouble seeing the numbers. How do you keep the writing dark? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Cathy Mayton Sent: Tuesday, May 11, 2004 10:08 AM To: Histonet Subject: [Histonet] marking blocks and slides Good morning, We use Sharpie brand industrial fine point marker to write on our blocks and slides. The marker goes through the processing and staining with no problems. These markers can be bought at office supply stores or maybe even at your local Wal-Mart, KMart etc. and are less expensive. Hope this helps. Cathy ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ "Quality Histology with a Personal Touch" A GLP Compliant Laboratory Cathy A. Mayton Wasatch Histo Consultants, Inc. 775-625-4425 www.wasatchhisto.com ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***** 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tflore <@t> lsuhsc.edu Tue May 11 11:05:34 2004 From: tflore <@t> lsuhsc.edu (Flores, Teresa) Date: Fri Sep 16 15:22:56 2005 Subject: [Histonet] Techs on Renal Biopsies Message-ID: Vicki, our EM lab is a diagnostic Electron Microscopy referral lab for several surrounding institutions/hospitals performing renal biopsies. One particular institution is a very aggressive Transplant Team (renal, liver and pancreases) and therefore most of their biopsies are transplant and require STAT results, same day results, which our lab provides if the biopsy is received before 11am. The clinician, the doctor performing the biopsy, in our case, it is usually the Nephrologists (sometimes if the biopsy will be difficult, a Radiologist is consulted, but strictly by them) will obtain the biopsy and place the core(s) in Paraformaldehyde-G Fixative for HRLM-TEM and Immunofluorescent Transport Media (both provided by us upon request). Institution sending biopsy also provides the courier. My pathologist goal, Dr. E. O. Hoffmann, has been to have the physician performing the biopsy to view the cores using a light microscope or a magnifying loop as glomerulus's can be seen and therefore be able to return to biopsy patient at bedside, if the biopsy is inadequate. I hope this helps you a bit. Teresa Flores LSUHSC New Orleans, LA -----Original Message----- From: Vicki Gauch [mailto:GauchV@mail.amc.edu] Sent: Tuesday, May 11, 2004 8:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Techs on Renal Biopsies I was just wondering how you all handle renal biopsy cases. Do your histotechs go on the biopsies to identify glomeruli, adequacy of tissue,and divide tissue for routine histology, immunofluorescence and electron microscopy studies, etc. or, if not the techs, who handles that in your institution? 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 From tflore <@t> lsuhsc.edu Tue May 11 11:12:48 2004 From: tflore <@t> lsuhsc.edu (Flores, Teresa) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Techs on Renal Biopsies Message-ID: Linda, so your techs actually have time to go to the patient bedside, to assist the doctor performing a biopsy, to see if the biopsy is adequate, sufficient glomerulus's, or are you speaking about technical work performed by the Histotechnologist. It really amazes me that given the shortage of Histotechnologist, this is possible. Teresa Flores LSUHSC New Orleans, LA -----Original Message----- From: Sebree Linda A. [mailto:la.sebree@hosp.wisc.edu] Sent: Tuesday, May 11, 2004 9:31 AM To: Vicki Gauch; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Techs on Renal Biopsies Hi Vicki, We have a renal lab staffed by a histologist that only does this type of work. She uses hisotechs from the surgical pathology lab as back-ups when she's busy or absent. They (surg. path. techs) train extensively with the renal histotech to be able to function as back-ups. 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 -----Original Message----- From: Vicki Gauch [mailto:GauchV@mail.amc.edu] Sent: Tuesday, May 11, 2004 8:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Techs on Renal Biopsies I was just wondering how you all handle renal biopsy cases. Do your histotechs go on the biopsies to identify glomeruli, adequacy of tissue,and divide tissue for routine histology, immunofluorescence and electron microscopy studies, etc. or, if not the techs, who handles that in your institution? 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Tue May 11 11:34:52 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Dilemma..... Message-ID: <494304423C63E246A5CF87A3AEEB577011B5ED@bumail01.barrynet.barry.edu> The old Gomori method for alkaline phosphatase left a an indestructible black deposit of cobalt sulfide. Gomori's method always gave me good results. The method is given in Lillie's HISTOPATHOLOGIC TECHNIC AND PRACTICAL HISTOCHEMISTRY. The 4th edition is out of print and unavailable, but a used copy of the 2nd or 3rd edition can be had from Barnes and Noble or Amazon at a reasonable price. The current (3rd) edition of Kiernan's HISTOLOGICAL AND HISTOCHEMICAL METHODS gives a cerium-DAB-nickel method that leaves a quite durable precipitate. Either method allows clearing in xylene and mounting in resin. I don't know how either would affect subsequent immunohistochemical reactions. 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 brucea@unimelb.edu.au Sent: Monday, May 10, 2004 9:59 PM To: histonet-bounces@lists.utsouthwestern.edu; histonet@lists.utsouthwestern.edu Subject: [Histonet] Dilemma..... Hi everyone I posted a question on the histonet a week ago but I don't think I explained my dilemma clearly enough. I am using alkaline phosphatase histochemistry (not immunohistochemistry!) on PFA-fixed whole marsupial blastocysts and embryos to detect primordial germ cells (PGCs). I use 1mg/ml Fast Blue salt and 1mg/ml Naphthol AS phosphate sodium salt in 0.2M Tris buffer, pH 9.4 for the histochemistry, a standard method for the detection of PGCs. The staining is good but I have been told that it is not permanent. Ideally, after staining these embryos histochemically for PGCs, I would like to dehydrate, paraffin- embed, section and then stain them immunohistochemically (ie, with antibodies) for other germ cell markers and several growth factors. Does anyone know of a histochemical method for the detection of germ cells that would later permit me to do this? We have well-established methods for immunohistochemistry for several germ cell markers in our lab. Thank you to the people who took the time to help last time, Dr Danielle Hickford Research Fellow Department of Zoology, University of Melbourne, Australia 3010. hickford@unimelb.edu.au -- BRUCE ABALOZ PH:61383446282 HISTOLOGIST FAX:61383447909 DEPT.of ZOOLOGY EMAIL: brucea@unimelb.edu.au THE UNIVERSITY Of MELBOURNE. VICTORIA.AUSTRALIA 3010 Nobody can make you feel inferior without your permission. - Eleanor Roosevelt DANCE LIKE NO-ONE'S WATCHING **************************************************************************** **** This electronic message and all contents contain information which may be privileged, confidential or otherwise protected from disclosure.The information is intended to be for the addressee only. If you are not the addressee, any disclosure, copy, distribution or use of the contents of this message is prohibited. If you have received this electronic message in error, please notify us immediately and destroy the original message and all copies. _______________________________________________ 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 pxs76 <@t> po.cwru.edu Tue May 11 12:08:42 2004 From: pxs76 <@t> po.cwru.edu (phyllis) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Processing of Mouse and Hamster Brains Message-ID: <5.1.0.14.0.20040511130343.03c88c80@pop.cwru.edu> We have recently started processing mouse and hamster brains for histology and immunohistochemistry. Can anyone share their processing schedule and also handling of tissue prior to processing. The tissue is brittle and when the microwe is used for the immnos the sections fall off. Phyllis Scalzo, HT(ASCP) Case Western Reserve University 2085 Adelbert Rd. Cleveland, Ohio 44106 Ph: 216-368-0822 Fax: 216-368-2546 From drmoses <@t> erols.com Tue May 11 13:33:45 2004 From: drmoses <@t> erols.com (david moses) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Ventana's Confirm CD10? Message-ID: <008f01c43786$7f0fe530$b3f13ad0@turbo> I am working up Ventana's Confirm CD10 antibody.The germinal centers of the normal tonsil are very pale, the follicular lymphoma is a little better and the kidney staining is good. I'm using Trilogy(EDTA) in a Decloaker for 4 min. at 120c, then a 10 min. cool down. Any suggestions on how I can improve the staining in my lymphoid tissue? Thanks. Renee Boston-Moses HT From GauchV <@t> mail.amc.edu Tue May 11 13:31:23 2004 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Thank you Message-ID: I just wanted to thank everyone who responded to my question regarding renal biopsy cases. I really appreciated the input since we are looking at our current methods for these cases. We send techs on the biopsies now and they are responsible for determining adequacy of tissue, division of tissue and presence of glomeruli. Our pathologist asked me to see what other institutions are doing which is where you all came in !!! I have already forwarded the info on so we'll see if anything changes now.... Have a wonderful day and thanks again, Vicki From CBWright <@t> nmcsd.med.navy.mil Tue May 11 14:14:42 2004 From: CBWright <@t> nmcsd.med.navy.mil (Wright, Clarissa B CIV) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Ventana's Confirm CD10? Message-ID: <4D840F2F36912841BD8B2484E8C6839303F96C14@nmc-sdca-exch02.med.navy.mil> David, How long is your primary incubation period? Are you using a Ventana automatic stainer? If so which one? C. Wright HT -----Original Message----- From: david moses [mailto:drmoses@erols.com] Sent: Tuesday, May 11, 2004 11:34 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Ventana's Confirm CD10? I am working up Ventana's Confirm CD10 antibody.The germinal centers of the normal tonsil are very pale, the follicular lymphoma is a little better and the kidney staining is good. I'm using Trilogy(EDTA) in a Decloaker for 4 min. at 120c, then a 10 min. cool down. Any suggestions on how I can improve the staining in my lymphoid tissue? Thanks. Renee Boston-Moses HT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This document may contain information covered under the Privacy Act, 5 USC 552(a), Health Insurance Portability and Accountability Act, Public Law 104-191, and DoD Directive 6025.18. It must be protected in accordance with those provisions. From funderwood <@t> mcohio.org Tue May 11 14:31:27 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] marking blocks and slides, Cathy Mayton Message-ID: Try Moist Mark Plus or HistoTec Pen. Both are available from Newcomer Supply. 1.800.383.7799 Fred >>> "Michelle Becker" 05/11/04 12:07PM >>> Have you tried Statmark Pens from StatLab Medical Products? We have had very good luck with them after trying several pens. It writes dark and stays on through both staining and processing procedures. They also last much longer than secureline markers. michb -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joyce Cline Sent: Tuesday, May 11, 2004 10:39 AM To: Histonet Subject: RE: [Histonet] marking blocks and slides, Cathy Mayton I have tried this and the writing was so light we had trouble seeing the numbers. How do you keep the writing dark? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Cathy Mayton Sent: Tuesday, May 11, 2004 10:08 AM To: Histonet Subject: [Histonet] marking blocks and slides Good morning, We use Sharpie brand industrial fine point marker to write on our blocks and slides. The marker goes through the processing and staining with no problems. These markers can be bought at office supply stores or maybe even at your local Wal-Mart, KMart etc. and are less expensive. Hope this helps. Cathy ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ "Quality Histology with a Personal Touch" A GLP Compliant Laboratory Cathy A. Mayton Wasatch Histo Consultants, Inc. 775-625-4425 www.wasatchhisto.com ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***** 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. _______________________________________________ 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 MAUGER <@t> email.chop.edu Tue May 11 14:39:00 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Ventana's Confirm CD10? Message-ID: Hi Renee! We are your old budies from HUP, now at CHOP! We use CD10 monoclonal from Santa Cruz. I know you're stuck with Ventana, so try a higher pH buffer for retreival. We use Dako's pH 9.5 for CD10 at 1:25. A CLL tonsil or LN is a good control. Rita and Carol, and me, Joanne, say hi and how are you? How are the twins? Good luck, Jo Mauger >>> "david moses" 05/11/04 02:33PM >>> I am working up Ventana's Confirm CD10 antibody.The germinal centers of the normal tonsil are very pale, the follicular lymphoma is a little better and the kidney staining is good. I'm using Trilogy(EDTA) in a Decloaker for 4 min. at 120c, then a 10 min. cool down. Any suggestions on how I can improve the staining in my lymphoid tissue? Thanks. Renee Boston-Moses HT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rolf.Brekken <@t> UTSouthwestern.edu Tue May 11 16:46:41 2004 From: Rolf.Brekken <@t> UTSouthwestern.edu (Rolf Brekken) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Re: Histonet Digest, Vol 6, Issue 16 (Out of Office reply) Message-ID: I am out of the lab until Monday May 24. I'll return your message as soon as I can after then. thanks rolf >>> histonet 05/11/04 16:46 >>> 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..." From neuroant <@t> hotmail.com Tue May 11 17:06:29 2004 From: neuroant <@t> hotmail.com (Ant Swensson) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] marking blocks and slides Message-ID: We were using Statmark pens for a while (admittedly before I arrived here) and were having a great experience with them, until we got a pen that came completely off in the processing. Since then, I can't get anyone to use a pen (although I have proved by using both pen and pencil on blocks that the pens are ok) and am stuck using a pencil. I'm suprised how the Sharpie (which I can remove from just about anything by using alcohol) can survive processing. I'll have to test it out. Antoinette Swensson Univeristy of Washington/Harborview Medical Center Neuropathology 325 9th Ave MS 359-791 Seattle, WA 98104 (206) 731-3910 >From: "Cathy Mayton" >To: "Histonet" >Subject: [Histonet] marking blocks and slides >Date: Tue, 11 May 2004 07:07:31 -0700 > >Good morning, > >We use Sharpie brand industrial fine point marker to write on our blocks and slides. The marker goes through the processing and staining with no problems. These markers can be bought at office supply stores or maybe even at your local Wal-Mart, KMart etc. and are less expensive. > >Hope this helps. >Cathy >^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ >"Quality Histology with a Personal Touch" >A GLP Compliant Laboratory > >Cathy A. Mayton >Wasatch Histo Consultants, Inc. >775-625-4425 >www.wasatchhisto.com >^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ [1]FREE pop-up blocking with the new MSN Toolbar get it now! References 1. http://g.msn.com/8HMBENUS/2728??PS=47575 From ploykasek <@t> phenopath.com Tue May 11 17:48:58 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Slide stainer Message-ID: Thanks to all for the replies about the linear "bicycle chain" stainer. I appreciate it. I was actually asking for a fellow researcher who is elsewhere in the city. Patti Loykasek PhenoPath Laboratories From GREYTRUNK <@t> aol.com Tue May 11 18:40:53 2004 From: GREYTRUNK <@t> aol.com (GREYTRUNK@aol.com) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Breast processing Message-ID: <50.2be29c24.2dd2be85@aol.com> Just an FYI---- All of those labs that is using the FDA approved protocol for Her2 should only be using formalin on the processors and for fixation. We use the following protocol and it works great. 10% NBF 2 hours 10% NBF 2 hours 70% ethanol 45 minutes 80% ethanol 45 minutes 95% ethanol 45 minutes 100% ethanol 45 minutes 100% ethanol 45 minutes 100% ethanol/xylene mix 45 minutes xylene 45 minutes xylene 45 minutes paraplast 30 minutes paraplast 30 minutes paraplast 30 minutes paraplast 30 minutes This protocol works great on fatty tissue as well, but again the section size must be reasonable ( no more than 4 mm) Roxanne From bhewlett <@t> cogeco.ca Tue May 11 19:51:06 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Breast processing References: <50.2be29c24.2dd2be85@aol.com> Message-ID: <000c01c437bb$35897770$6500a8c0@mainbox> Just another FYI----- Fixation of breast tissue for the IHC demonstration of HER2 requires at least 16-24 hours in NBF. 4 hours in NBF on the processor comes nowhere near the FDA approved protocol for HER2, unless your tissue has already spent at least 12 hours in NBF prior to being placed on the processor!! Bryan ----- Original Message ----- From: To: ; ; Sent: Tuesday, May 11, 2004 7:40 PM Subject: Re: [Histonet] Breast processing > Just an FYI---- > All of those labs that is using the FDA approved protocol for Her2 should > only be using formalin on the processors and for fixation. We use the following > protocol and it works great. > > 10% NBF 2 hours > 10% NBF 2 hours > 70% ethanol 45 minutes > 80% ethanol 45 minutes > 95% ethanol 45 minutes > 100% ethanol 45 minutes > 100% ethanol 45 minutes > 100% ethanol/xylene mix 45 minutes > xylene 45 minutes > xylene 45 minutes > paraplast 30 minutes > paraplast 30 minutes > paraplast 30 minutes > paraplast 30 minutes > > This protocol works great on fatty tissue as well, but again the section size > must be reasonable ( no more than 4 mm) > Roxanne > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From laurie.reilly <@t> jcu.edu.au Wed May 12 02:08:58 2004 From: laurie.reilly <@t> jcu.edu.au (Laurie Reilly) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Breast processing In-Reply-To: <27066863AD02214B81FE49477F3E71AD0D6F2C42@hinet1.hinet.org> Message-ID: <5.1.0.14.0.20040512170604.00a43610@mail.jcu.edu.au> Subject: RE: Processing Fatty Tissues To: HistoNet The major problem with processing fatty tissues, assuming that they are fixed properly, is that Ethanol is not a good solvent for fat and therefore cannot penetrate the tissues completely, so the tissues are inadequately dehydrated. We have had some success with lipomas by adding a "degreasing" step of xylene into the processing schedule. 70% ethanol 80% ethanol 90% ethanol 95% ethanol Absolute ethanol Xylene Absolute ethanol Xylene Xylene Paraffin Paraffin Paraffin The first Absolute ethanol will dehydrate the tissue to some extent. The next Xylene step will remove most of the fat and then the second Absolute ethanol can complete the dehydration. A compromise situation that we use routinely is to have Absolute ethanol 50:50 Absolute ethanol:Xylene Xylene This is not quite as effective but it is less disruptive to the normal schedule and handles moderately fatty tissues. Laurie Reilly Histopathology Phone 077 81 4468 Australian Institute of Tropical Veterinary and Animal Science Fax 077 81 5558 James Cook University Townsville. 4811 Australia. At 01:18 PM 05/10/04 -0700, Bliss, Mary E. wrote: >Hi All, > > > >How do other laboratorians prepare fatty breast specimens for >histology? Are you doing anything special? We had a case today which >our doctor needs to have re-processed. It appears unfixed, although it >sat in formalin on the processor over the weekend before it processed on >Sunday night. The sections are large (too large in my opinion) and not >adequately removed of fat. We are using toluene on our processors and >have considered going to Xylene. We have tried Penn fixx in the past, >but discontinued using it years ago. I know it is a complicated >subject, but just thought I would see if anyone has any bits of wisdom. > > > > > >Mary E. Bliss > >Lead Histologist > >Northwest Pathology, P.S. > >3614 Meridian St. Suite 100 > >Bellingham, WA 98225 > >(360)734-2800 x601 > >(360)734-3818 FAX > > > > > >_______________________________________________ >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 hmcleod <@t> chempath.uct.ac.za Wed May 12 03:39:56 2004 From: hmcleod <@t> chempath.uct.ac.za (Mcleod) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] complement Message-ID: <40A1E2DC.73349D1D@chempath.uct.ac.za> Hi All What are the pitfalls of staining complement on FFPE material ... if any? Would Histonetters who stain regularly for complement kindly share their expertise. I and a master's student will really appreciate any help. Thanks in advance, Heather From hmcleod <@t> chempath.uct.ac.za Wed May 12 04:13:41 2004 From: hmcleod <@t> chempath.uct.ac.za (Mcleod) Date: Fri Sep 16 15:22:57 2005 Subject: [Fwd: [Histonet] complement] Message-ID: <40A1EAC5.A415C5BD@chempath.uct.ac.za> I forgot to mention that we will be working on SKIN tissue. Thanks Heather From balajimr <@t> drreddys.com Wed May 12 04:24:42 2004 From: balajimr <@t> drreddys.com (balajimr@drreddys.com) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Rta Brain - processing Message-ID: Can somebody suggest me a best processing for rat brains. Recently I am having the problem of cracks and tares in the brain sections. We are collecting the sections at 40 degree c. Any suggestions are welcome. Thanks in advance. Following is my processing schedule: Station No. Solvent Time (Hr) 1 Isopropyl Alcohol 80% 1 2 Isopropyl Alcohol 90% 1 3 Isopropyl Alcohol 95% 1 4 Isopropyl Alcohol 95% 1 5 Isopropyl Alcohol 100% 1 6 Isopropyl Alcohol 100% 1 7 Isopropyl Alcohol 100% 1 8 Chloroform 1 9 Chloroform 1 10 Chloroform 1 11 Paraffin I 3 12 Paraffin II 3 From c.m.vanderloos <@t> amc.uva.nl Wed May 12 04:24:42 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] RE: Cox -2 IHC Message-ID: <790d8f793226.793226790d8f@amc.uva.nl> Dear Moumita Kundu, If the original protocol for this COX-2 antibody was set up with a TSA detection kit (and was working) there is a good chance that detection of this antibody with neither LSAB+ nor LSAB2 will show any specific positive staining. Simply because those detection systems are far less sensitive/efficient. My message is to test both detection systems and compare your results. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From "Moumita Kundu" Date Tue, 11 May 2004 10:56:04 -0500 To "Elizabeth Chlipala" , "'Histonet'" Subject [Histonet] Cox -2 IHC I am trying to do IHC with Cox -2 ,The protocol I have is a long overnight incubation with TSA kit. I am trying to reduce the time of incubation using Dako's LSAB+ system or LSAB2 . Please pass me any protocol which can be helpful. Thank you in advance. From marktarango <@t> earthlink.net Wed May 12 05:26:40 2004 From: marktarango <@t> earthlink.net (Mark Adam Tarango) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Breast processing In-Reply-To: <200405111001.1bnADp5vB3NZFjK0@condor> Message-ID: <000001c4380b$9d477c40$2702a8c0@markie> Well, I've seen many people take a piece of fatty breast tissue while embedding and squish/squash/squeeze it until all that juicy fat just drips out. After this, they let it sit in molten paraffin for a few minutes. I don't recommend this, I only mention it hoping someone will say what a horrible practice this is. It seems like squeezing breasts is the norm lol . . . maybe this really is normal, but I don't like the idea of it. . . Any comments.....? Mark Tarango -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bliss, Mary E. Sent: Monday, May 10, 2004 4:19 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Breast processing Hi All, How do other laboratorians prepare fatty breast specimens for histology? Are you doing anything special? We had a case today which our doctor needs to have re-processed. It appears unfixed, although it sat in formalin on the processor over the weekend before it processed on Sunday night. The sections are large (too large in my opinion) and not adequately removed of fat. We are using toluene on our processors and have considered going to Xylene. We have tried Penn fixx in the past, but discontinued using it years ago. I know it is a complicated subject, but just thought I would see if anyone has any bits of wisdom. Mary E. Bliss Lead Histologist Northwest Pathology, P.S. 3614 Meridian St. Suite 100 Bellingham, WA 98225 (360)734-2800 x601 (360)734-3818 FAX From sharon.willman <@t> bms.com Wed May 12 08:14:34 2004 From: sharon.willman <@t> bms.com (Sharon E Willman) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Nuclear (DNA) Stain Message-ID: <40A2233A.2BB47656@bms.com> Hi, I was wondering what is the best special stain used for staining nucleus (DNA) in tissue that has been fixed in Bouin's. Any help would be most appreciated. Thanks in advance! Sharon Willman From Janet.Bonner <@t> FLHOSP.ORG Wed May 12 09:43:11 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Alternative's to the Steiner Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB3FC1@fh2k093.fhmis.net> Speaking of the Steiner, is there a substitute solution that can be used instead of 1% Uranyl Nitrate? I would appreciate any help - Thank you. Janet -----Original Message----- From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] Sent: Tuesday, May 11, 2004 8:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Alternative's to the Steiner I have been given alternatives's to stain Helicobacter pylori, however, what about the other organisms stained by the Steiner method? -spirochetes -legionella -donavan bodies -cat scratch Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 _______________________________________________ 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 mcauliff <@t> umdnj.edu Wed May 12 12:17:27 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Nuclear (DNA) Stain In-Reply-To: <40A2233A.2BB47656@bms.com> References: <40A2233A.2BB47656@bms.com> Message-ID: <40A25C27.9020304@umdnj.edu> The Feulgen reaction (Schiff's reagent after HCl hydrolysis) comes to mind. Geoff Sharon E Willman wrote: >Hi, >I was wondering what is the best special stain used for staining >nucleus (DNA) in tissue that has been fixed in Bouin's. Any >help would be most appreciated. > >Thanks in advance! > >Sharon Willman > > >_______________________________________________ >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 juan.gutierrez <@t> christushealth.org Wed May 12 09:49:06 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Alternative's to the Steiner Message-ID: I'm not sure about the Steiner, but we used phosphomolybdic acid one time with our Dieterle stain. The concentration eludes me, but I think it was something like 5 or 10 percent. Good luck. Juan C. Gutierrez, HT(ASCP) Histology Supervisor Christus Santa Rosa Hospital 333 N. Santa Rosa Ave. San Antonio, TX 78207 -----Original Message----- From: Bonner, Janet [mailto:Janet.Bonner@FLHOSP.ORG] Sent: Wednesday, May 12, 2004 9:43 AM To: 'Mary Reeves'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Alternative's to the Steiner Speaking of the Steiner, is there a substitute solution that can be used instead of 1% Uranyl Nitrate? I would appreciate any help - Thank you. Janet -----Original Message----- From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] Sent: Tuesday, May 11, 2004 8:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Alternative's to the Steiner I have been given alternatives's to stain Helicobacter pylori, however, what about the other organisms stained by the Steiner method? -spirochetes -legionella -donavan bodies -cat scratch Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 _______________________________________________ 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 From HornHV <@t> archildrens.org Wed May 12 09:46:12 2004 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Breast processing Message-ID: Thank you Bryan for pointing this out. As a BC survivor it means a lot to me to know there are labs who are following proper protocol. I understand it's difficult to get pathologists to wait the proper time for fixation of breast tissue but techs must demand this. You can bet if it was their wife they would insist on the tissue being fixed longer. 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 Bryan Hewlett Sent: Tuesday, May 11, 2004 7:51 PM To: GREYTRUNK@aol.com; browning@HHSC.CA; mary.bliss@northwestpathology.com; histonet@pathology.swmed.edu Subject: Re: [Histonet] Breast processing Just another FYI----- Fixation of breast tissue for the IHC demonstration of HER2 requires at least 16-24 hours in NBF. 4 hours in NBF on the processor comes nowhere near the FDA approved protocol for HER2, unless your tissue has already spent at least 12 hours in NBF prior to being placed on the processor!! Bryan ----- Original Message ----- From: To: ; ; Sent: Tuesday, May 11, 2004 7:40 PM Subject: Re: [Histonet] Breast processing > Just an FYI---- > All of those labs that is using the FDA approved protocol for Her2 > should only be using formalin on the processors and for fixation. We > use the following > protocol and it works great. > > 10% NBF 2 hours > 10% NBF 2 hours > 70% ethanol 45 minutes > 80% ethanol 45 minutes > 95% ethanol 45 minutes > 100% ethanol 45 minutes > 100% ethanol 45 minutes > 100% ethanol/xylene mix 45 minutes > xylene 45 minutes > xylene 45 minutes > paraplast 30 minutes > paraplast 30 minutes > paraplast 30 minutes > paraplast 30 minutes > > This protocol works great on fatty tissue as well, but again the > section size > must be reasonable ( no more than 4 mm) > Roxanne > _______________________________________________ > 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 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 mfredrickson <@t> cohenderm.com Wed May 12 09:40:49 2004 From: mfredrickson <@t> cohenderm.com (Michael Fredrickson) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Immunoperoxidase on FFPE for Lyme disease Message-ID: <3D55809F621E7C438DA80098CDE3744D10C4CD@hub.cohenderm.com> The season is starting for Lyme disease. Is anyone performing immunostaining to demonstrate B. burgdoferi on FFPE tissue? Mike Fredrickson Technical Director Cohen Dermatopathology, PC Newton, MA 02464 T (617) 969-4100 F (617) 969-3393 mfredrickson@cohenderm.com From juan.gutierrez <@t> christushealth.org Wed May 12 09:39:09 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Ventana's Confirm CD10? Message-ID: Hello Renee, we use Ventana's CD10 on the Benchmark with the standard CC1, A/B block, and amplification with great results accross the board. Hope this helps. Juan C. Gutierrez, HT(ASCP) Histology Supervisor Christus Santa Rosa Hospital 333 N. Santa Rosa Ave. San Antonio, TX 78207 -----Original Message----- From: david moses [mailto:drmoses@erols.com] Sent: Tuesday, May 11, 2004 1:34 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Ventana's Confirm CD10? I am working up Ventana's Confirm CD10 antibody.The germinal centers of the normal tonsil are very pale, the follicular lymphoma is a little better and the kidney staining is good. I'm using Trilogy(EDTA) in a Decloaker for 4 min. at 120c, then a 10 min. cool down. Any suggestions on how I can improve the staining in my lymphoid tissue? Thanks. Renee Boston-Moses HT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SnyderW <@t> uhcwv.org Wed May 12 09:52:38 2004 From: SnyderW <@t> uhcwv.org (Snyder, Wendy) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Frozen sections Message-ID: <7B5CC54F6E30BD42A8C61E4896484D19011A9079@uhc-exchange.uhcwv.org> Hello, I was wondering about a policy for the length of time a frozen section block can safely remain in a cryostat before it is taken out and routinely processed. I have a pathologist that wants to leave the frozen tissue block in the cryostat for up to 3 days in case he wants to do immuno stains from it. I would be interested to here of other policys from other institutions regarding this issue. Thank you, Wendy Snyder United Hospital Center Clarksburg, WV From ian.montgomery <@t> bio.gla.ac.uk Wed May 12 10:33:14 2004 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Comments. Message-ID: <6.1.0.6.2.20040512162529.02cfd6a0@udcf.gla.ac.uk> Just been staining rat oesophagus with PGP9.5 for nerves, worked a treat. Interestingly a small piece of thyroid and parathyroid has been left with the oesophagus. While the Chief cells in the parathyroid have no staining the Oxyphil cells display a strong reaction product. Surprised by this result. Anyone like to comment? Ian. 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 Rcartun <@t> harthosp.org Wed May 12 10:26:33 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Immunoperoxidase on FFPE for Lyme disease Message-ID: Yes, I do IHC for Borrelia burgdorferi. Although we have been successful in detecting the spirochete in animal tissue, I have never seen a positive human tissue. Therefore, I'm not sure I would recommend doing it. Richard Cartun Director, Immunopathology Hartford Hospital Hartford, CT 06102 >>> "Michael Fredrickson" 05/12/04 10:40AM >>> The season is starting for Lyme disease. Is anyone performing immunostaining to demonstrate B. burgdoferi on FFPE tissue? Mike Fredrickson Technical Director Cohen Dermatopathology, PC Newton, MA 02464 T (617) 969-4100 F (617) 969-3393 mfredrickson@cohenderm.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Wed May 12 10:26:50 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Breast processing Message-ID: " You can bet if it was their wife they would insist on the tissue being fixed longer." I doubt it. Most pathologist and some techs from this list, think that a tissue is fixed when it's not bloody any more. 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: Horn, Hazel V [mailto:HornHV@archildrens.org] Sent: 12 May 2004 15:46 To: Bryan Hewlett; GREYTRUNK@aol.com; browning@HHSC.CA; mary.bliss@northwestpathology.com; histonet@pathology.swmed.edu Subject: RE: [Histonet] Breast processing Thank you Bryan for pointing this out. As a BC survivor it means a lot to me to know there are labs who are following proper protocol. I understand it's difficult to get pathologists to wait the proper time for fixation of breast tissue but techs must demand this. You can bet if it was their wife they would insist on the tissue being fixed longer. 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 Bryan Hewlett Sent: Tuesday, May 11, 2004 7:51 PM To: GREYTRUNK@aol.com; browning@HHSC.CA; mary.bliss@northwestpathology.com; histonet@pathology.swmed.edu Subject: Re: [Histonet] Breast processing Just another FYI----- Fixation of breast tissue for the IHC demonstration of HER2 requires at least 16-24 hours in NBF. 4 hours in NBF on the processor comes nowhere near the FDA approved protocol for HER2, unless your tissue has already spent at least 12 hours in NBF prior to being placed on the processor!! Bryan ----- Original Message ----- From: To: ; ; Sent: Tuesday, May 11, 2004 7:40 PM Subject: Re: [Histonet] Breast processing > Just an FYI---- > All of those labs that is using the FDA approved protocol for Her2 > should only be using formalin on the processors and for fixation. We > use the following > protocol and it works great. > > 10% NBF 2 hours > 10% NBF 2 hours > 70% ethanol 45 minutes > 80% ethanol 45 minutes > 95% ethanol 45 minutes > 100% ethanol 45 minutes > 100% ethanol 45 minutes > 100% ethanol/xylene mix 45 minutes > xylene 45 minutes > xylene 45 minutes > paraplast 30 minutes > paraplast 30 minutes > paraplast 30 minutes > paraplast 30 minutes > > This protocol works great on fatty tissue as well, but again the > section size > must be reasonable ( no more than 4 mm) > Roxanne > _______________________________________________ > 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 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GauchV <@t> mail.amc.edu Wed May 12 10:07:38 2004 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Frozen sections Message-ID: Wendy, We submit our frozen section blocks from the OR as soon as the frozen is diagnosed. The block is removed from the cryostat and placed in a cassette in a bin of formalin with our other specimens that are being grossed in the OR that day. Those cases are all processed that night and cut as surgicals the following morning. The exceptions to that rule are renal biopsies and skin biopsies that are specifically submitted for IMF- those are kept frozen for long periods of time after the IMF has been completed. Vicki Gauch AMCH Albany, NY >>> "Snyder, Wendy" 5/12/2004 10:52:38 AM >>> Hello, I was wondering about a policy for the length of time a frozen section block can safely remain in a cryostat before it is taken out and routinely processed. I have a pathologist that wants to leave the frozen tissue block in the cryostat for up to 3 days in case he wants to do immuno stains from it. I would be interested to here of other policys from other institutions regarding this issue. Thank you, Wendy Snyder United Hospital Center Clarksburg, WV _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jenny-Oblander <@t> omrf.ouhsc.edu Wed May 12 10:05:06 2004 From: Jenny-Oblander <@t> omrf.ouhsc.edu (Jenny Oblander) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Nuclear (DNA) Stain Message-ID: BioPath has an excellent Feulgen , they have reduced the time to 2 hours rather then 3, which helped me out. BioPath phone # 1-800-728-4462 ask to speak with Lee Angros, he will answer any questions you might have. Jenny -----Original Message----- From: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] Sent: Wednesday, May 12, 2004 12:17 PM To: Sharon E Willman Cc: histonet@pathology.swmed.edu Subject: Re: [Histonet] Nuclear (DNA) Stain The Feulgen reaction (Schiff's reagent after HCl hydrolysis) comes to mind. Geoff Sharon E Willman wrote: >Hi, >I was wondering what is the best special stain used for staining >nucleus (DNA) in tissue that has been fixed in Bouin's. Any >help would be most appreciated. > >Thanks in advance! > >Sharon Willman > > >_______________________________________________ >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 ********************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kgrobert <@t> rci.rutgers.edu Wed May 12 11:09:06 2004 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Grocott's Stain...not my day today Message-ID: <40A24C22.9000903@rci.rutgers.edu> Well, I decided to try looking into the borax first, in part because I found out that the person who made it up for me used boric acid rather than sodium tetraborate (we didn't have any of that, hence the boric acid). So I got the sodium tetraborate from Sigma (cat no. 221732) this morning, and went to make up the 5% borax solution.... it won't go into solution. :::sigh::: There is nothing in the AFIP protocol about making it up, other than "5g borax in 100 ml distilled water". Any ideas or suggestions? I tried heating it a little bit, but it didn't budge. Should I try to shift the pH? Return this stuff and get one of the decahydrates? (If you do a search for "Borax" on Sigma's web site, you get a whole slew of items.) Thanks in advance for any and all advice. :o) Feeling like I am losing my touch- Kathleen Roberts Neurotoxicology Labs Rutgers University From lcardenas <@t> utmem.edu Wed May 12 11:12:15 2004 From: lcardenas <@t> utmem.edu (Luz Cardenas) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] cytochrome oxidase Message-ID: Hi everyone, 1.Can cytochrome oxidase method stain fixed tissue (4% Paraformaldehyde)? 2. Is there any specific method for fixed tissue? Thank you very much in advance! Luz. Luz C?rdenas, MD Department of Anatomy and Neurobiology University of Tennessee Health Science Center 855 Monroe Avenue, Room 309/302 Memphis, TN 38163 Lab: (901) 448-6002 / 7318 Fax: (901) 448-7193 (Dept) lcardenas@utmem.edu From RossS <@t> BaylorHealth.edu Wed May 12 10:45:35 2004 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Frozen sections Message-ID: I suspect that if it stays frozen it will be fine. The problem with leaving it in the cryostat is the defrost cycle. So it isn't staying frozen. I think it would be best kept in a -70 freezer. I will be interested to see other opinions. 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 Snyder, Wendy Sent: Wednesday, May 12, 2004 9:53 AM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] Frozen sections Hello, I was wondering about a policy for the length of time a frozen section block can safely remain in a cryostat before it is taken out and routinely processed. I have a pathologist that wants to leave the frozen tissue block in the cryostat for up to 3 days in case he wants to do immuno stains from it. I would be interested to here of other policys from other institutions regarding this issue. Thank you, Wendy Snyder United Hospital Center Clarksburg, WV _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From gcallis <@t> montana.edu Wed May 12 11:27:11 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Sodium borate aka borax, GMS and Kathleen Roberts Message-ID: <3.0.6.32.20040512102711.00c001d8@gemini.msu.montana.edu> The 5% sodium borate is not supposed to go into solution totally, it is a SATURATED solution and when you use it, just pipette off the top. Your day is NOT ruined. 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 TJJ <@t> Stowers-Institute.org Wed May 12 11:03:21 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Re: breast processing Message-ID: Mark, although I recall seeing this published in the Journal of Histotechnology, I also don't agree with teh idea of squeezing the fat out of fatty breast tissue. Other labs have also just soaked them in molten paraffin overnight, sometimes resulting in burned looking tissue. If they are not adequately fixed and processed well in the first place, any of this type of trauma can't be good for the specimen. I do like the idea of adding a xylene step in the processing schedule between the alcohols, never thought of that. Brilliant! I do suggest that some labs that have particular difficulty with their pathologists routinely not grossing in these specimens well, use this as a Quality Improvement marker. I would talk with the Lab Director and Head Pathologist about doing this first because you will need their support, but that might help supply the pressure (from somewhere besides the techs) to improve their technique. Bottom line is, we do what is best for the patient. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From MAUGER <@t> email.chop.edu Wed May 12 11:00:15 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Frozen sections Message-ID: Wendy, Storing tissue in the cryostat is a bad idea because it has an automatic defrost cycle for prevention of ice build up. The tissue will thaw and refreeze some, leading to freezing artifact-like holes in tissue, etc. Morphology suffers. I suggest cutting extra slides and keeping them frozen until staining, or store the tissue in a freezer temporarily. Jo >>> "Vicki Gauch" 05/12/04 11:07AM >>> Wendy, We submit our frozen section blocks from the OR as soon as the frozen is diagnosed. The block is removed from the cryostat and placed in a cassette in a bin of formalin with our other specimens that are being grossed in the OR that day. Those cases are all processed that night and cut as surgicals the following morning. The exceptions to that rule are renal biopsies and skin biopsies that are specifically submitted for IMF- those are kept frozen for long periods of time after the IMF has been completed. Vicki Gauch AMCH Albany, NY >>> "Snyder, Wendy" 5/12/2004 10:52:38 AM >>> Hello, I was wondering about a policy for the length of time a frozen section block can safely remain in a cryostat before it is taken out and routinely processed. I have a pathologist that wants to leave the frozen tissue block in the cryostat for up to 3 days in case he wants to do immuno stains from it. I would be interested to here of other policys from other institutions regarding this issue. Thank you, Wendy Snyder United Hospital Center Clarksburg, WV _______________________________________________ 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 TJJ <@t> Stowers-Institute.org Wed May 12 10:53:17 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Re: Frozen sections Message-ID: Wendy, Because of the "thaw" schedule of most cryostats, storing a block of tissue in the cryostat overnight is not a good practice. If your pathologist is worried about losing orientation of the tissue (or whatever the rationale is...) keep the sample on the chuck, wrap it in foil, and store it in a -20 to -80C freezer overnight. If you use the -20C freezer, make absolutely certain it is not a frost-free model. Otherwise you can cut your slides in advance and store the remainder of unused slides in the freezer and take out what is necessary when you need them. 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 Wed May 12 11:32:07 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Rat brain processing Message-ID: <3.0.6.32.20040512103207.00c001d8@gemini.msu.montana.edu> You did not say if you are processing whole brains, bisected brains OR coronal slices of brains. If you are doing the latter, 3 mm slices, your schedule is too long and you are overprocessing the tissues which creates your sectioning problems. Shorten the processing schedule will help this problem. You will see a great deal of discussion on this recently found in Histonet Archives, do a keyword search. Too much dehydration, clearing and heat of paraffin is drying out the tissue. You can try soaking the trimmed block in ice water to help sectioning, but it is tedious. 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 Sue.Kapoor <@t> uhsi.org Wed May 12 11:32:18 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] billing question Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F55D9@khmcexch.uhsi.org> when we have a specimen for frozen section, our doc's bill for a frozen, and then also bill for routine processing...is this correct? Thank you, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From gcallis <@t> montana.edu Wed May 12 11:45:21 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Feulgen stain for DNA Message-ID: <3.0.6.32.20040512104521.00c001d8@gemini.msu.montana.edu> For Feulgens, Bouins may not be the best fixative particularly if you fix a long time in this solution. Acetic acid in Bouins the acetic acid in Bouins starts hydrolyzing the purine and pyrimidine bases from sugar phosphate groups to release aldhyde groups from the DNA deoxypentose sugar. This is the very reason acid decalcifiers are not recommended if one wants to do Feulgens. When you then do Feulgens, HCL continues this hydrolyzation, the problem lies in overhydrolyzation, and poor staining. Formalin may be a better fixation. 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 nmuvarak <@t> facstaff.wisc.edu Wed May 12 11:35:59 2004 From: nmuvarak <@t> facstaff.wisc.edu (NIDAL E MUVARAK) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Fixing frozen sections in 4% Paraformaldehyde. Message-ID: <28870a286898.28689828870a@wiscmail.wisc.edu> Is it common to fix frozen sections of unfixed tissue with 4% PFA the same way they can be fixed with acetone? I know it will definitely take longer than aceton fixation, but has that been done? Thanks. Nidal E Muvarak Associate Research Specialist Vascular Tissue Biomechanics Laboratory Department of Biomedical Engineering University of Wisconsin-Madison 1550 Engineering Dr.; Rm. 2158 Madison, WI 53706-1609 Lab: (608) 265-8921; Office: (608) 265-4205; Home: (608) 256-7934; Cell: (608) 332-6068 http://vtb.bme.wisc.edu From ALAbeyta <@t> salud.unm.edu Wed May 12 11:37:04 2004 From: ALAbeyta <@t> salud.unm.edu (Antonia Abeyta) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Frozen sections Message-ID: I agree, we keep some of our tissue at -20, which is roughly where the cryostat stays, but I would worry about the defrost, refreezing. As a rule we avoid this by cutting everything at once. We had some tissue in the past that we were shipping back and forth across the county in dry ice and some of the tissue thawed and was then put back in the freezer, when stained the morphology was horrible, we ended up having to throw out data from that tissue! Maybe think about storing it in a -20 freezer on the block so it isn't so hard to reface if your pathologist needs more sections. Antonia >>> "Stapf, Ross" 05/12/04 09:45AM >>> I suspect that if it stays frozen it will be fine. The problem with leaving it in the cryostat is the defrost cycle. So it isn't staying frozen. I think it would be best kept in a -70 freezer. I will be interested to see other opinions. 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 Snyder, Wendy Sent: Wednesday, May 12, 2004 9:53 AM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] Frozen sections Hello, I was wondering about a policy for the length of time a frozen section block can safely remain in a cryostat before it is taken out and routinely processed. I have a pathologist that wants to leave the frozen tissue block in the cryostat for up to 3 days in case he wants to do immuno stains from it. I would be interested to here of other policys from other institutions regarding this issue. Thank you, Wendy Snyder United Hospital Center Clarksburg, WV _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Wed May 12 11:58:45 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Breast processing Message-ID: You may consider using a dehydrating solution which is 7 parts absolute alcohol and 3 parts xylene. I use 6 changes on my processer then clear with xylene. I don't process anything quite as fatty as beef steaks of breast tissue, but do run a variety of rather thickly grossed tissue. Fred >>> "Mark Adam Tarango" 05/12/04 06:26AM >>> Well, I've seen many people take a piece of fatty breast tissue while embedding and squish/squash/squeeze it until all that juicy fat just drips out. After this, they let it sit in molten paraffin for a few minutes. I don't recommend this, I only mention it hoping someone will say what a horrible practice this is. It seems like squeezing breasts is the norm lol . . . maybe this really is normal, but I don't like the idea of it. . . Any comments.....? Mark Tarango -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bliss, Mary E. Sent: Monday, May 10, 2004 4:19 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Breast processing Hi All, How do other laboratorians prepare fatty breast specimens for histology? Are you doing anything special? We had a case today which our doctor needs to have re-processed. It appears unfixed, although it sat in formalin on the processor over the weekend before it processed on Sunday night. The sections are large (too large in my opinion) and not adequately removed of fat. We are using toluene on our processors and have considered going to Xylene. We have tried Penn fixx in the past, but discontinued using it years ago. I know it is a complicated subject, but just thought I would see if anyone has any bits of wisdom. Mary E. Bliss Lead Histologist Northwest Pathology, P.S. 3614 Meridian St. Suite 100 Bellingham, WA 98225 (360)734-2800 x601 (360)734-3818 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Wed May 12 12:03:59 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] billing question Message-ID: Yes, there are specific CPT codes for intraoperative consultations in addition to those used for routine surgical pathology specimens. Richard Cartun >>> "Kapoor, Sue" 05/12/04 12:32PM >>> when we have a specimen for frozen section, our doc's bill for a frozen, and then also bill for routine processing...is this correct? Thank you, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GauchV <@t> mail.amc.edu Wed May 12 12:06:40 2004 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Summary of Renal Biopsy Handling Responses Message-ID: Peggy asked if I would share the information I received on how institutions handled renal biopsies-as far as who determined adequacy of tissue, presence of glomeruli and division of tissue for IMF,EM and routine histology. This is what I received: 4 - Pathologists do the determinations AND also divide the tissue. 3 - Technical staff do the determinations AND the division of tissue. 1 - Technical staff go on the biopsy but the pathologist makes the determinations and divides the tissue. 1 - Renal Lab Tech does the determinations and division of tissue. 1 - Surgeon/Radiologist do the determination and division of tissue. 1 - Surgeon/Radiologist brings the tissue to the pathologist who makes the determinations and division of tissue. 2 - Pathologist and technical staff - the pathologist makes the determinations and the tech divides the tissue. At my institution the tech goes on the biopsy, makes the determination and also divides the tissue. Hope you find this information useful.. Have a nice day, Vicki From BlazekL <@t> childrensdayton.org Wed May 12 10:42:37 2004 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Frozen sections Message-ID: If your cryostat is set up to defrost at night it would be a very bad idea to leave the tissue in the cryostat. 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 >>> "Snyder, Wendy" 5/12/2004 10:52:38 AM >>> Hello, I was wondering about a policy for the length of time a frozen section block can safely remain in a cryostat before it is taken out and routinely processed. I have a pathologist that wants to leave the frozen tissue block in the cryostat for up to 3 days in case he wants to do immuno stains from it. I would be interested to here of other policys from other institutions regarding this issue. Thank you, Wendy Snyder United Hospital Center Clarksburg, WV _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Wed May 12 12:22:42 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Re: breast processing Message-ID: In my recent past, I worked with an excellent pathologist who would thinly (5-10 cm) "breadloaf" mastectomy and other large breast specimens, placing paper towels between each slice to wick in the formalin. These specimens would fix at least overnight in large closed 'tupperware' vats before additional grossing early in the a.m. We would then fix the cassettes in alcoholic formalin for the rest of the day (6-7 hours) before routine processing. (No need to go back into 10% NBF - started off in 70% on the processor. Specimens like this were processed for 1 hour each station (under vacuum), and I don't recall having any underprocessed/underfixed tissues. Don't tell the kiddies, but in my more distant past, when I did come across a block that was underprocessed, I'd take a good whiff to see what solution (alcohol, xylene, formalin) I smelled in the block, and back it up to that point only. If I smelled xylene, it needed more time in paraffin - plopping it back into a couple of fresh changes of paraffin for a couple of hours seemed to do the trick. If it smelled like alcohol, it was backed up to fresh xylene for a couple of hours, then brought forward again. Ahh, the good old days. Thanks Flonase. Jackie O' "Johnson, Teri" Sent by: histonet-bounces@lists.utsouthwestern.edu 05/12/2004 11:03 AM To: cc: histonet@pathology.swmed.edu Subject: [Histonet] Re: breast processing Mark, although I recall seeing this published in the Journal of Histotechnology, I also don't agree with teh idea of squeezing the fat out of fatty breast tissue. Other labs have also just soaked them in molten paraffin overnight, sometimes resulting in burned looking tissue. If they are not adequately fixed and processed well in the first place, any of this type of trauma can't be good for the specimen. I do like the idea of adding a xylene step in the processing schedule between the alcohols, never thought of that. Brilliant! I do suggest that some labs that have particular difficulty with their pathologists routinely not grossing in these specimens well, use this as a Quality Improvement marker. I would talk with the Lab Director and Head Pathologist about doing this first because you will need their support, but that might help supply the pressure (from somewhere besides the techs) to improve their technique. Bottom line is, we do what is best for the patient. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Wed May 12 15:37:55 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] cytochrome oxidase In-Reply-To: References: Message-ID: <40A28B23.6050204@umdnj.edu> Dear Luz: Luz Cardenas wrote: >Hi everyone, >1.Can cytochrome oxidase method stain fixed tissue (4% Paraformaldehyde)? > Yes, provided the fixation is short. Very short. Fix fresh frozen sections 5 min in cold buffered formalin. Whole animal perfusion for 15 seconds followed by a buffered sucrose washout in preparation for frozen sections. >2. Is there any specific method for fixed tissue? > Lots of published methods. See John Kiernan's book (which you should have in the lab) or papers by Margaret Wong-Riley. I don't know if the "standard" histopath texts would have this method, perhaps too specialized? Geoff > >Thank you very much in advance! >Luz. > >Luz C?rdenas, MD >Department of Anatomy and Neurobiology >University of Tennessee Health Science Center >855 Monroe Avenue, Room 309/302 >Memphis, TN 38163 >Lab: (901) 448-6002 / 7318 >Fax: (901) 448-7193 (Dept) >lcardenas@utmem.edu > > >_______________________________________________ >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 WesterM <@t> MedImmune.com Wed May 12 12:45:58 2004 From: WesterM <@t> MedImmune.com (Wester, Martha) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] RE: Fixing frozens in 4% PFA Message-ID: <83899F0EC7671543B305FB5694024DE6BAD302@medimmune4.medimmune.com> I frequently use an immunohistochemistry protocol which calls for BRIEF fixation of the frozen slides in PFA or 10% Neutral buffered Formalin (30sec up to 5min). This seems to help adhere the sections to the slide more so than have any additive effect on the tissue. I 've never played with the fixation times to see at what point IHC signal is affected. Good Luck! Martha Wester Associate Scientist MedImmune, Inc. One MedImmune Way Gaithersburg, MD 20878 301-398-4794 fax 301-398-9794 Message: 13 Date: Wed, 12 May 2004 11:35:59 -0500 From: NIDAL E MUVARAK Subject: [Histonet] Fixing frozen sections in 4% Paraformaldehyde. To: histonet@pathology.swmed.edu Message-ID: <28870a286898.28689828870a@wiscmail.wisc.edu> Content-Type: text/plain; charset=us-ascii Is it common to fix frozen sections of unfixed tissue with 4% PFA the same way they can be fixed with acetone? I know it will definitely take longer than aceton fixation, but has that been done? Thanks. Nidal E Muvarak Associate Research Specialist Vascular Tissue Biomechanics Laboratory Department of Biomedical Engineering University of Wisconsin-Madison 1550 Engineering Dr.; Rm. 2158 Madison, WI 53706-1609 Lab: (608) 265-8921; Office: (608) 265-4205; Home: (608) 256-7934; Cell: (608) 332-6068 http://vtb.bme.wisc.edu From convmcm <@t> cc.usu.edu Wed May 12 12:45:27 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Cool websites Message-ID: <000601c43848$e90890f0$4a737b81@Cygnus> Hi everyone! I just found two really fun websites you all might enjoy. 1. "Bizarre stuff you can make in your kitchen" For those of you who have children in your home, this site is awesome (I consider myself to be a rather large child, so age isn't important) http://home.houston.rr.com/molerat/2nddef.htm 2. Anyone familiar with Roitt's book, Immunology, will find added attractions to that book on this website: http://www.fleshandbones.com/immunology/roitt Enjoy!! Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 From JPCOLEMA <@t> sentara.com Wed May 12 13:02:43 2004 From: JPCOLEMA <@t> sentara.com (JOHN COLEMAN) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] CD10, CD103, FDA, fatty tissues, complement, frozen blocks, fixation Message-ID: For CD10 we use Novo Castra's antibody at a 1:30 dilution, Trilogy from Cellmarque (high pH, edta, tween) for 15 minutes in a 98C water bath, and biogenex's super sensitive concentrate kit for detection. 45 minute primary antibody incubation. Question- I need a reliable CD103 antibody for formalin fixed paraffin embedded material. The clone we have currently is good on coagulative fixed and air dried material, but is worthless on routinely proccessed bone marrow and lymphoid tissue. Pardon my cynical response, but FDA doesn't care if it works. If you follow the protocol you're in. If you tweak the protcol you're not in FDA land and can not claim such, even if your controls look great. For fatty tissues we have a fixative that is 4 gal formalin,1 gal absolute ETOH and 500 ml Glacial acetic acid . We place the blocks in this after the tissue is grossed in at the board. Complement- we stain for C3c, C4c, and C1q on FFPE kidney and skin biopsies using .1% protease from sigma for 30 minutes @ 40C before DIF labelling. Does your cryostat have a defrost cycle? if so the blocks are being trashed. I recommend sealing in oct, wrapping in parafilm and storing in a -70 freezer or processing as usual and doing immunos on formalin fixed tissue, where the morphology is better. My 2 cents on fixation- tissue is fixed when it's fixed.(Formalin penetrates and fixes 2mm per hour , right?) It's not fixed because the pathologist wants the tissue out early. I think the sixteen hours is overkill myself and IHC retrieval methods are in place to counteract long fixation. If it were my wife I would cut the tissue in at 4 mm, fix for 8 hours and run the tests like I do with all my patients. I've optimized my lab to give every single person the best possible result. My wife or daughter or mother is no more or less valuable than anyone else that walks through the door. If anyone would do differently for people important to them in their lab then you guys need to bring your standards up! This is medicine- someone's life litereally depends on our results. If your routine methods are not good enough for your family you should be ashamed to leave them in place. From fmonson <@t> wcupa.edu Wed May 12 13:06:55 2004 From: fmonson <@t> wcupa.edu (Monson, Frederick ) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Comments. Message-ID: Comments are inexpensive, but invited comments may be of even less value/cost. Interesting that the cell (Oxyphil) about which we know little is loaded with mitochondria, replicates at a reasonable rate ( http://jcem.endojournals.org/cgi/reprint/82/8/2681.pdf ) and demonstrates, on Ian's slide, and in contrast to its sister cell, the "Chief cell," an immunohistochemical link to the diffuse neuroendocrine system. Perhaps the poor little Oxyphil is playing an esoteric part in the local regulatory processes. The story of 'Ca' regulation as it is explained these days is complicated but still likely to be an oversimplification of the reality. Perhaps one day the Oxyphil will change its name to the "Super-chief cell." http://www.qu.edu.qa/myqu/nasserizk/downloadfiles/463.pdf An old story tells of PTH-stimulated Ca release from a piece of bone taken from an 'old bone drawer' in an anatomy lab. The story tells of a concurrently run control bone fragment that didn't leak, so it can't be disparaged on experimental grounds on that point. Cheers to all, P.S. My first look at rodent Sertoli cells came while I perused a section of turtle ovary and paused for a minute mammalian (RBC's) aside. Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone/FAX: 610-738-0437 eMail: fmonson@wcupa.edu CASI Page and Scheduling http://darwin.wcupa.edu/CASI/ -----Original Message----- From: Ian Montgomery [mailto:ian.montgomery@bio.gla.ac.uk] Sent: Wednesday, May 12, 2004 11:33 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Comments. Just been staining rat oesophagus with PGP9.5 for nerves, worked a treat. Interestingly a small piece of thyroid and parathyroid has been left with the oesophagus. While the Chief cells in the parathyroid have no staining the Oxyphil cells display a strong reaction product. Surprised by this result. Anyone like to comment? Ian. 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 gcallis <@t> montana.edu Wed May 12 13:23:17 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Fixing frozen sections in 4% Paraformaldehyde. In-Reply-To: <28870a286898.28689828870a@wiscmail.wisc.edu> Message-ID: <3.0.6.32.20040512122317.00bfd728@gemini.msu.montana.edu> We prefer to do a time study with 4% paraformaldehyde - cold at 4C. 2,4,6,8,10 min. and have done the same with cold 4C acetone. Some antigens can withstand longer fixation in PFA and/or acetone, while picky antigens require shorter time. Frozen sections are air dried at RT, then immersed all at the same time, with a section pulled at specific time point, rinsed with Dulbeccos PBS x 3. It is work, but in the long run has paid off rather than go back and adjust fixation time. At 11:35 AM 5/12/2004 -0500, you wrote: >Is it common to fix frozen sections of unfixed tissue with 4% PFA the same way they can be fixed with acetone? I know it will definitely take longer than aceton fixation, but has that been done? Thanks. > >Nidal E Muvarak >Associate Research Specialist >Vascular Tissue Biomechanics Laboratory >Department of Biomedical Engineering >University of Wisconsin-Madison >1550 Engineering Dr.; Rm. 2158 >Madison, WI 53706-1609 >Lab: (608) 265-8921; Office: (608) 265-4205; >Home: (608) 256-7934; Cell: (608) 332-6068 >http://vtb.bme.wisc.edu > > >_______________________________________________ >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 info <@t> instrumedics.com Wed May 12 13:35:57 2004 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] frozen block in cryostat Message-ID: <020301c4384f$f747b4a0$6401a8c0@INSTRUMEDICS22> Wendy, I think that in three days the tissue and the embedding medium will begin to dehydrate. Instrumedics Protective Oil is designed to coat the block without melting the surface of the block and will prevent dehydration. Please visit our web site for the details www.instrumedics.com If you have any questions please contact me. Bernice ----- Original Message ----- From: "Snyder, Wendy" To: Sent: Wednesday, May 12, 2004 10:52 AM Subject: [Histonet] Frozen sections > Hello, > I was wondering about a policy for the length of time a frozen section > block can safely remain in a cryostat before it is taken out and routinely > processed. I have a pathologist that wants to leave the frozen tissue block > in the cryostat for up to 3 days in case he wants to do immuno stains from > it. I would be interested to here of other policys from other institutions > regarding this issue. > > Thank you, > Wendy Snyder > United Hospital Center > Clarksburg, WV > From dlcowie <@t> prodigy.net Wed May 12 13:41:35 2004 From: dlcowie <@t> prodigy.net (Dawn Cowie) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] microwave processing Message-ID: <20040512184135.14038.qmail@web81003.mail.yahoo.com> hi histonetters, Is anyone using the RHS-1 microwave tissue processor with vacuum? If so, would you care to share with me the processing schedule you use for tiny bx's such as gastric, liver cores, etc. We have not been having great success so far. Any help would be appreciated. Thanks, Dawn Cowie HT From kwittle <@t> jhmi.edu Wed May 12 13:49:35 2004 From: kwittle <@t> jhmi.edu (Karen Wittler) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Borax,GMS Message-ID: I think you want Borax decahydrate Na2B4O7-10H2O named "Sodium tetraborate decahydrate", in the Sigma catalog. Also, the borax does not totally dissolve, it's a saturated solution. Karen Wittler, HT(ASCP) Johns Hopkins Hospital Baltimore, MD From marytedo <@t> hotmail.com Wed May 12 14:10:26 2004 From: marytedo <@t> hotmail.com (María Teresa Domínguez) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Breast processing Message-ID: Well, here I use this for the breast processing in the Tissue's processor: 10% NBF x 2 jar,2 hours in each 70% Ethanol x1 j, 1 hour and a half 96% Alcohol x 3 jar ,1 hour and a half in each Acetone x 1 j,1 hour and a half Methil Alcohol x 1 jar,1 hour and a half N-Buthylic Acetate x 1 jar, 1 hour and a half Xileno x 1 jar,1 hour and a half Parafine 56ºC- 58ºC x 2 jar,2 hours and a half in each Embedding, and cut the pieces... That's all for me. [i.p.emwink.gif] Ht. Maria Teresa Dominguez Anatomic Pathology Service Hospital Regional Río Grande, Tierra del Fuego, Argentina. From: "Connie McManus" >To: "'Bliss, Mary E.'" , >Subject: RE: [Histonet] Breast processing >Date: Mon, 10 May 2004 15:31:07 -0600 > >Mary, my heart goes out to you. I believe every histotech in the world >has gone through this same nightmare --- tissues that are too big and >not properly processed, and the pathologist expects YOU, the histotech, >to produce perfect results. > > >From your story, fixation may not be the only problem. 48 hours is >generally plenty of time for tissues to fix. But if there is a lot of >fatty tissue and not enough room for the fixative to work through the >tissue (as in a tissue smushed up inside a processing cassette), you may >have a fixation problem compounded with a processing problem. The only >solution is for the person trimming in the tissues to keep them between >2 and 3 mm in thickness and to try to keep the length and width >dimensions to something less than that of the cassette. There should >absolutely not be any tissue sticking out of the cassette nor should >there be imprints of the cassette on the tissue. > >As for the fat in your tissues, if there was adequate processing, it >would be gone. Alcohols, toluene, xylene and xyl substitutes do a good >job of that. > >There's nothing quite like good reagent flow in and around the tissue >while processing ... > >Connie McManus >Utah Veterinary Diagnostics Laboratory >Utah State University >Logan, UT >Phone: 435/797-1891 >fax: 435/797-2805 > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bliss, >Mary E. >Sent: Monday, May 10, 2004 1:19 PM >To: histonet@pathology.swmed.edu >Subject: [Histonet] Breast processing > >Hi All, > > > >How do other laboratorians prepare fatty breast specimens for >histology? Are you doing anything special? We had a case today which >our doctor needs to have re-processed. It appears unfixed, although it >sat in formalin on the processor over the weekend before it processed on >Sunday night. The sections are large (too large in my opinion) and not >adequately removed of fat. We are using toluene on our processors and >have considered going to Xylene. We have tried Penn fixx in the past, >but discontinued using it years ago. I know it is a complicated >subject, but just thought I would see if anyone has any bits of wisdom. > > > > > >Mary E. Bliss > >Lead Histologist > >Northwest Pathology, P.S. > >3614 Meridian St. Suite 100 > >Bellingham, WA 98225 > >(360)734-2800 x601 > >(360)734-3818 FAX > > > > > >_______________________________________________ >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 _________________________________________________________________ [1]Best Restaurant Giveaway Ever! Vote for your favorites for a chance to win $1 million! References 1. http://g.msn.com/8HMBENUS/2746??PS=47575 From REEVEML <@t> shands.ufl.edu Wed May 12 14:17:41 2004 From: REEVEML <@t> shands.ufl.edu (Mary Reeves) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] microwave processing Message-ID: We use the RHS-2 for our small biopsies (no vacuum) and have had wonderful success. I am pretty sure you can run your RHS-1 without the vacuum. It is a computer programming change a service tech from Hacker would need to make. Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 >>> Dawn Cowie 05/12/04 02:41PM >>> hi histonetters, Is anyone using the RHS-1 microwave tissue processor with vacuum? If so, would you care to share with me the processing schedule you use for tiny bx's such as gastric, liver cores, etc. We have not been having great success so far. Any help would be appreciated. Thanks, Dawn Cowie HT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Angel.Enniss <@t> pfizer.com Wed May 12 14:23:41 2004 From: Angel.Enniss <@t> pfizer.com (Enniss, Angel) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] CAB Stain Message-ID: <4E2F83D5DEE32C41B4816FE9D63462420B3CC4C0@anagrdexm01.research.aa.wl.com> I am new to the CAB stain and have been experimenting with it. I have found bits and pieces of literature on the stain and its origination. If I could get any information that could be helpful, it would be gratefully appreciated!! Kindest Regards, Angel C. Enniss Angel.Enniss@Pfizer.com _____ Upgrade Your Email - Click here! 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 tbourm <@t> olympicmedical.org Wed May 12 14:59:32 2004 From: tbourm <@t> olympicmedical.org (Tasha Bourm) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] NAIL CLIPPINGS Message-ID: CAN SOMEONE SHARE WITH ME THE BEST WAY TO PROCESS NAIL CLIPPINGS. I WILL BE DOING A PAS STAIN FOR FUNGI THANKS TASHA BOURM From convmcm <@t> cc.usu.edu Wed May 12 14:59:03 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Cool websites In-Reply-To: <000601c43848$e90890f0$4a737b81@Cygnus> Message-ID: <000b01c4385b$93090dc0$4a737b81@Cygnus> Folks, I don't know what the deal is with the Bizarre kitchen stuff website. It worked just fine when I tried it at home last night and again this AM when I had to sneak a peak at it again. It doesn't work any more. Did I break it????? =:0 Seriously, I appologize to anyone who tried it and didn't get any of the links to work. Try again later -- it's well worth it. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Connie McManus Sent: Wednesday, May 12, 2004 10:45 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Cool websites Hi everyone! I just found two really fun websites you all might enjoy. 1. "Bizarre stuff you can make in your kitchen" For those of you who have children in your home, this site is awesome (I consider myself to be a rather large child, so age isn't important) http://home.houston.rr.com/molerat/2nddef.htm 2. Anyone familiar with Roitt's book, Immunology, will find added attractions to that book on this website: http://www.fleshandbones.com/immunology/roitt Enjoy!! Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Wed May 12 15:15:30 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Feulgen stain for DNA Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3B30@exchsrv01.barrynet.barry.edu> Bouin's certainly does attack nucleotides, often making the Feulgen reaction impossible (although sometimes one can get a Feulgen reaction on Bouin-fixed tissue by just plopping it into Schiff's reagent). I think it is the picric acid, pK 0.4, rather than the acetic acid, pK 4.8, that hydrolyzes the nucleotides. Carnoy's fluid, with twice the acetic acid concentration of Bouin's, is a good fixative for the Feulgen reaction. 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 Gayle Callis Sent: Wednesday, May 12, 2004 12:45 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Feulgen stain for DNA For Feulgens, Bouins may not be the best fixative particularly if you fix a long time in this solution. Acetic acid in Bouins the acetic acid in Bouins starts hydrolyzing the purine and pyrimidine bases from sugar phosphate groups to release aldhyde groups from the DNA deoxypentose sugar. This is the very reason acid decalcifiers are not recommended if one wants to do Feulgens. When you then do Feulgens, HCL continues this hydrolyzation, the problem lies in overhydrolyzation, and poor staining. Formalin may be a better fixation. 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 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 Dixon.Leslie <@t> mayo.edu Wed May 12 16:00:20 2004 From: Dixon.Leslie <@t> mayo.edu (Dixon, Leslie E.) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Cool websites Message-ID: <438DAC78076DD611A3FD0002B39F7F683DBA51@excsrv21.mayo.edu> It works if you just use this address http://home.houston.rr.com/molerat/ -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Wednesday, May 12, 2004 12:59 PM To: 'Connie McManus'; histonet@pathology.swmed.edu Subject: RE: [Histonet] Cool websites Folks, I don't know what the deal is with the Bizarre kitchen stuff website. It worked just fine when I tried it at home last night and again this AM when I had to sneak a peak at it again. It doesn't work any more. Did I break it????? =:0 Seriously, I appologize to anyone who tried it and didn't get any of the links to work. Try again later -- it's well worth it. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Connie McManus Sent: Wednesday, May 12, 2004 10:45 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Cool websites Hi everyone! I just found two really fun websites you all might enjoy. 1. "Bizarre stuff you can make in your kitchen" For those of you who have children in your home, this site is awesome (I consider myself to be a rather large child, so age isn't important) http://home.houston.rr.com/molerat/2nddef.htm 2. Anyone familiar with Roitt's book, Immunology, will find added attractions to that book on this website: http://www.fleshandbones.com/immunology/roitt Enjoy!! Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 _______________________________________________ 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 asmith <@t> mail.barry.edu Wed May 12 16:25:42 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Nuclear (DNA) Stain Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3B31@exchsrv01.barrynet.barry.edu> Methyl green does not work after Bouin fixation. Sometimes the Feulgen reaction does. If you have enough tissue, try several different hydrolysis times, e.g., 0 min., 2 min., 4 min., 6 min. 0.1% toluidine blue at pH 4.2 will usually stain RNA violet and DNA indigo, even after Bouin fixation. Kernechtrot, alum-brazilin, or Ehrlich's hematoxylin stain nuclei very well after Bouin fixation, but they stain nucleoproteins rather than DNA as such. 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 Sharon E Willman Sent: Wednesday, May 12, 2004 9:15 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Nuclear (DNA) Stain Hi, I was wondering what is the best special stain used for staining nucleus (DNA) in tissue that has been fixed in Bouin's. Any help would be most appreciated. Thanks in advance! Sharon Willman _______________________________________________ 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 bhewlett <@t> cogeco.ca Wed May 12 19:32:28 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] CD10, CD103, FDA, fatty tissues, complement, frozen blocks, fixation References: <20040512180929.C45582519@fep4.cogeco.net> Message-ID: <000401c43881$c5c56080$6500a8c0@mainbox> John wrote: >>My 2 cents on fixation- tissue is fixed when it's fixed.(Formalin penetrates and fixes 2mm per hour , right?) It's not fixed because the pathologist wants the tissue out early. I think the sixteen hours is overkill myself and IHC retrieval methods are in place to counteract long fixation. If it were my wife I would cut the tissue in at 4 mm, fix for 8 hours and run the tests like I do with all my patients. I've optimized my lab to give every single person the best possible result. >> The penetration rate of formaldehyde is largely irrelevant to tissue no more than 1.5 cm thick. However, it's NOT 2mm per hour and fixation takes much longer than this! It's variable. d =K x the square root of time. K = the coefficient of diffusibility. Baker determined K=3.6, probably correct and a figure most people accept. Helander's 1994 data suggests it is at least 2.0 and most likely close to 3.6. Using K=3.6 in the above formula, the first layer of cells is penetrated at a rate of over 90 mm per hour. This slows to 3.6 mm at 1 hour. It then takes 4 hours to double the depth penetrated to 7.2 mm, a rate of 1.8mm per hour. At 16 hours, the depth is again doubled to 14.4 mm, a rate of 0.9 mm/hr. i.e. to double the depth of penetration you must allow 4X the time and so doing halves the rate! Your 4mm tissue block would be fully penetrated in less than 1 hour (3.6 mm from each surface) What the issue really is, relates to the binding time necessary for formaldehyde. A 90% threshold binding, i.e. MINIMAL binding time is 24 hours at 22C and 18 hours at 37C. It doesn't matter if your tissue is only a few cells thick or 4mm thick! 16 hours is not long fixation, 3 weeks may be, and is certainly NOT overkill. The weak initial binding of formaldehyde is freely reversed, 50% in about 12 hours, by the lower alcohols on the processor and the tissue becomes fixed by alcohol. Any thing less than 8 hours formaldehyde fixation reduces the IHC for ER demonstrably and HER2 markedly. Bryan ----- Original Message ----- From: "JOHN COLEMAN" To: Sent: Wednesday, May 12, 2004 2:02 PM Subject: [Histonet] CD10, CD103, FDA, fatty tissues, complement,frozen blocks, fixation For CD10 we use Novo Castra's antibody at a 1:30 dilution, Trilogy from Cellmarque (high pH, edta, tween) for 15 minutes in a 98C water bath, and biogenex's super sensitive concentrate kit for detection. 45 minute primary antibody incubation. Question- I need a reliable CD103 antibody for formalin fixed paraffin embedded material. The clone we have currently is good on coagulative fixed and air dried material, but is worthless on routinely proccessed bone marrow and lymphoid tissue. Pardon my cynical response, but FDA doesn't care if it works. If you follow the protocol you're in. If you tweak the protcol you're not in FDA land and can not claim such, even if your controls look great. For fatty tissues we have a fixative that is 4 gal formalin,1 gal absolute ETOH and 500 ml Glacial acetic acid . We place the blocks in this after the tissue is grossed in at the board. Complement- we stain for C3c, C4c, and C1q on FFPE kidney and skin biopsies using .1% protease from sigma for 30 minutes @ 40C before DIF labelling. Does your cryostat have a defrost cycle? if so the blocks are being trashed. I recommend sealing in oct, wrapping in parafilm and storing in a -70 freezer or processing as usual and doing immunos on formalin fixed tissue, where the morphology is better. My 2 cents on fixation- tissue is fixed when it's fixed.(Formalin penetrates and fixes 2mm per hour , right?) It's not fixed because the pathologist wants the tissue out early. I think the sixteen hours is overkill myself and IHC retrieval methods are in place to counteract long fixation. If it were my wife I would cut the tissue in at 4 mm, fix for 8 hours and run the tests like I do with all my patients. I've optimized my lab to give every single person the best possible result. My wife or daughter or mother is no more or less valuable than anyone else that walks through the door. If anyone would do differently for people important to them in their lab then you guys need to bring your standards up! This is medicine- someone's life litereally depends on our results. If your routine methods are not good enough for your family you should be ashamed to leave them in place. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.m.vanderloos <@t> amc.uva.nl Thu May 13 01:52:31 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] RE: Frozen sections Message-ID: <66ab30671092.67109266ab30@amc.uva.nl> Dear Wendy, Apart from the damage to the tissue morphology caused by the defrost cycle of the cryostat that has been mentioned a couple of times, there is another risk. Water crystals tend to re-crystallize from amorphous to needles between -1 and -30C. Depending on the temperature this process takes a couple of days and causes holes in the tissue. The lower the temperature, the slower this process will be. And this is exactly the reason why we are storing our frozen tissue at -70 and -80C. Don't leave your tissue in the cryostat! Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From "Snyder, Wendy" Date Wed, 12 May 2004 10:52:38 -0400 To "'histonet@pathology.swmed.edu'" Subject [Histonet] Frozen sections Hello, I was wondering about a policy for the length of time a frozen section block can safely remain in a cryostat before it is taken out and routinely processed. I have a pathologist that wants to leave the frozen tissue block in the cryostat for up to 3 days in case he wants to do immuno stains from it. I would be interested to here of other policys from other institutions regarding this issue. Thank you, Wendy Snyder United Hospital Center Clarksburg, WV From ian.montgomery <@t> bio.gla.ac.uk Thu May 13 04:42:10 2004 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:22:57 2005 Subject: Fwd: RE: [Histonet] Comments. Message-ID: <6.1.0.6.2.20040513103624.02d17eb0@udcf.gla.ac.uk> Richard, Novocastra. Looking at the section again, comparing with adjacent H&E's and beginning to change my mind about Oxyphils. Nerves so deep into the gland, mmm, not sure yet. Will have a word with our Regis Professor of Physiology, who knows everything, well he was appointed by the Queen so he makes no mistakes. Will keep you informed. Ian. >Subject: RE: [Histonet] Comments. >Date: Thu, 13 May 2004 10:02:18 +0100 >X-MS-Has-Attach: >X-MS-TNEF-Correlator: >Thread-Topic: [Histonet] Comments. >Thread-Index: AcQ4NvDt6b+U12cKQCCKllSxu4sYIQAkeG8A >From: "Edwards, R.E." >To: "Ian Montgomery" >X-GLA-Spam-Scan: R >X-GLA-Spam-Score: 0.0 (/) >X-GLA-Spam-Report: > >Unable to answer your ????? but where do you get the PGP >9.5 from????.................thanks > >Richard Edwards > >MRC TOX UNIT > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Ian >Montgomery >Sent: 12 May 2004 16:33 >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Comments. > > > Just been staining rat oesophagus with PGP9.5 for nerves, worked a >treat. Interestingly a small piece of thyroid and parathyroid has been left >with the oesophagus. While the Chief cells in the parathyroid have no >staining the Oxyphil cells display a strong reaction product. Surprised by >this result. Anyone like to comment? >Ian. > >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 Rcartun <@t> harthosp.org Thu May 13 08:37:59 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] VEGF IHC & Bevacizumab Message-ID: Is anyone doing IHC for "vascular endothelial growth factor" (VEGF) to qualify patients with metastatic colorectal CA for treatment with Avastin (bevacizumab) or is it not needed? Thank you. Richard Cartun From Gldnsthisto <@t> aol.com Thu May 13 08:09:05 2004 From: Gldnsthisto <@t> aol.com (Gldnsthisto@aol.com) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Safety regs/Pathologists Message-ID: <1f1.20786cca.2dd4cd71@aol.com> Can anyone out there help me with what to do when the pathologists and their staff (not paid by the hospital) will not follow safety regulations. Thx From JWEEMS <@t> sjha.org Thu May 13 08:54:30 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Safety regs/Pathologists Message-ID: Let hospital administration know so that it can be addressed at contract time. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gldnsthisto@aol.com Sent: Thursday, May 13, 2004 9:09 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Safety regs/Pathologists Can anyone out there help me with what to do when the pathologists and their staff (not paid by the hospital) will not follow safety regulations. Thx _______________________________________________ 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 Joseph?s Health System, Inc. From Diane.Gladney <@t> se.amedd.army.mil Thu May 13 08:57:59 2004 From: Diane.Gladney <@t> se.amedd.army.mil (Gladney, Diane C Ms MACH) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Safety regs/Pathologists Message-ID: <9D41AB7C56F8304F98537ABD87B249F66261FC@DASMTHGBZ001> If you are a CAP Inspected lab, there is a requirement that all staff (even contract staff) are required to follow proper safety regulations/requirements. I'm sure that your hospital must have some safety regulations in place, also. If we do not follow the safety procedures, we are subject to dismissal. I strongly suggest that your lab manager get involved in this and get guidance from the hospital safety officer (or appropriate title). I'm sure that there are OSHA regulations also. I work in a military hospital so we have a lot of safety rules that are imposed on us by the Army, but I would think that any hospital would have mandatory safety procedures/rules in place and if they are not followed, then the individual is subject to discipline and/or dismissal. Thanks, Diane Diane C. Gladney, HT (ASCP) Histology /Cytology Supervisor Moncrief Army Community Hospital P.O. BOX 484 4500 Stuart Ave. FT. Jackson, SC 29207 (803) 751-2530 DSN 734-2530 EMAIL: diane.gladney@se.amedd.army.mil OR dcgx1@aol.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gldnsthisto@aol.com Sent: Thursday, May 13, 2004 9:09 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Safety regs/Pathologists Can anyone out there help me with what to do when the pathologists and their staff (not paid by the hospital) will not follow safety regulations. Thx _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pmarcum <@t> polysciences.com Thu May 13 09:05:20 2004 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Safety regs/Pathologists In-Reply-To: Message-ID: <002201c438f3$5390e670$4000a8c0@PMARCUM2K> Has your hospital management addressed this the group as possible reason for the laboratory having an issue if you are CAP inspected? OR the possibility that if they do not follow the rules the group could be replaced or censured at contract renewal time? Depending on exactly which rules they ignore they could be placing the hospital and staff in position for a lawsuit if someone is infected due to the negligence. Good Luck!! Thanks, Pam Marcum? > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Weems, > Joyce > Sent: Thursday, May 13, 2004 9:55 AM > To: 'Gldnsthisto@aol.com'; histonet@pathology.swmed.edu > Subject: RE: [Histonet] Safety regs/Pathologists > > > Let hospital administration know so that it can be addressed at contract > time. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of > Gldnsthisto@aol.com > Sent: Thursday, May 13, 2004 9:09 AM > To: histonet@pathology.swmed.edu > Subject: [Histonet] Safety regs/Pathologists > > > Can anyone out there help me with what to do when the pathologists and > their > > staff (not paid by the hospital) will not follow safety regulations. > Thx > _______________________________________________ > 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 Joseph?s Health System, Inc. > > From Jackie.O'Connor <@t> abbott.com Thu May 13 09:11:36 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Safety regs/Pathologists Message-ID: My husband is a corporate safety guru for Motorola, OSHA certified and all that junk. He tells me that the organization (in your case the hospital) is still liable for ensuring that contractors (the pathologists and their staff) follow appropriate safety rules and regs. It all depends on their contract - which we can only assume states that they WILL agree to follow established safety guidelines (to protect the hospital from liability lawsuits). In most cases, if they are not abiding by the conditions of their contract, the contract can be terminated. If I don't wear my safety glasses in the lab, MY boss's butt is on the line if one of his superiors sees me violating established rules. If you are in charge of this lab and enforcing safety guidelines, you are probably responsible for making sure everyone in your lab follows the rules. If this is the case, you should document their refusal, and go up the line until you find someone who has more clout, (such as the hospital safety officer or risk management) and document everyone you speak to regarding this issue. What kind of safety infractions are we talking about? Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Gldnsthisto@aol.com Sent by: histonet-bounces@lists.utsouthwestern.edu 05/13/2004 08:09 AM To: histonet@pathology.swmed.edu cc: Subject: [Histonet] Safety regs/Pathologists Can anyone out there help me with what to do when the pathologists and their staff (not paid by the hospital) will not follow safety regulations. Thx _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bill501 <@t> mindspring.com Thu May 13 09:19:18 2004 From: bill501 <@t> mindspring.com (Bill) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Safety regs/Pathologists In-Reply-To: <1f1.20786cca.2dd4cd71@aol.com> References: <1f1.20786cca.2dd4cd71@aol.com> Message-ID: At 9:09 AM -0400 5/13/04, Gldnsthisto@aol.com wrote: >Can anyone out there help me with what to do when the pathologists and their >staff (not paid by the hospital) will not follow safety regulations As a Pathologist and Physician, I try to never let regulations get in the way of my caring for patients. With 12 years of intensive education after high school and several decades of experience I claim, nay, demand the right to decide how to behave in a clinical setting for myself without interference of bureaucrats or rule-mongers. -- _____________________________ Bill http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board From Liam.Brennan <@t> bll.n-i.nhs.uk Thu May 13 09:45:50 2004 From: Liam.Brennan <@t> bll.n-i.nhs.uk (Brennan, Liam) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Safety regs/Pathologists Message-ID: "Work our way or work elsewhere!" -----Original Message----- From: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: 13 May 2004 14:55 To: 'Gldnsthisto@aol.com'; histonet@pathology.swmed.edu Subject: RE: [Histonet] Safety regs/Pathologists Let hospital administration know so that it can be addressed at contract time. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gldnsthisto@aol.com Sent: Thursday, May 13, 2004 9:09 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Safety regs/Pathologists Can anyone out there help me with what to do when the pathologists and their staff (not paid by the hospital) will not follow safety regulations. Thx _______________________________________________ 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 Joseph?s Health System, Inc. From kristenhinkle <@t> yahoo.com Thu May 13 10:13:11 2004 From: kristenhinkle <@t> yahoo.com (Kristen Reynolds) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Looking for job opportunities in Austin Message-ID: <20040513151311.3336.qmail@web41503.mail.yahoo.com> I am moving to Austin this August and am looking for a research position and/or histology position. I am HTL certified and do IHC on a daily basis. If you know of opportunities please let me know. Kristen Reynolds kristenhinkle@yahoo.com __________________________________ Do you Yahoo!? Yahoo! Movies - Buy advance tickets for 'Shrek 2' http://movies.yahoo.com/showtimes/movie?mid=1808405861 From tpmorken <@t> labvision.com Thu May 13 10:31:18 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Thick frozen section IHC Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA206EAD4@usca0082k08.labvision.apogent.com> Hi all, If you do IHC on thick (20+um) frozen sections, would mind sharing your procedures with me, escpecially concerning whether you do them mounted on slides, and if you use an automated IHC stainer. If you have a publication, or can direct me to one I would much appreciate it. Thanks! Tim Morken Lab Vision - Neomarkers www.labvision.com From Jackie.O'Connor <@t> abbott.com Thu May 13 10:11:53 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Histology bench ergonomics Message-ID: Does anyone have any data or information on bench height for microtomy? I know a lot of people cut at routine lab bench height (36" I think) with no trouble, and some cut at desk high benches (30"). Is there any safety/ergonomic guidelines or opinions as to which is better for microtomy? I actually prefer desk height, so I can keep my feet on the floor - I'm just wondering if there is anything official out there - I've looked at a couple of sites, but can't find a lot to support my idea that desk height is better. Thanks - Jackie O' From Stephen.J.Scholz <@t> osfhealthcare.org Thu May 13 10:56:26 2004 From: Stephen.J.Scholz <@t> osfhealthcare.org (Scholz, Stephen J.) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Looking for Elizabeth Wenig Message-ID: <7F1312711CA7474A89B3DF8BA0BA54D0F5F111@pmc-rfd-mx01.intranet.osfnet.org> Elizabeth Wenig, if you get this message please contact me. Stephen J. Scholz HT(ASCP) Histology Coordinator OSF St. Anthony Medical Center Rockford IL Phone: 815-395-5410 Fax: 815-395-5364 e-mail: sjscholz@osfhealthcare.org From Terry.Marshall <@t> rothgen.nhs.uk Thu May 13 10:55:21 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Safety regs/Pathologists Message-ID: Although Bill's views may sound rather prima donna-ish to some, I must support him. Safety procedures and practices have become rather petty, tedious, and in my view, sometimes more dangerous than the perceived danger designed for. Goggles prevent you seeing well - dangerous. Feather blade holders prevent you sensing where the blade is - dangerous. Safety hoods/cabinets prevent you seeing or doing anything in other than a clumsy way - dangerous, and so on. Let sense rule over nonsense. NB - this is NOT in any way anti-safety. 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: Bill [mailto:bill501@mindspring.com] Sent: 13 May 2004 15:19 To: histonet@pathology.swmed.edu Subject: Re: [Histonet] Safety regs/Pathologists At 9:09 AM -0400 5/13/04, Gldnsthisto@aol.com wrote: >Can anyone out there help me with what to do when the pathologists and their >staff (not paid by the hospital) will not follow safety regulations As a Pathologist and Physician, I try to never let regulations get in the way of my caring for patients. With 12 years of intensive education after high school and several decades of experience I claim, nay, demand the right to decide how to behave in a clinical setting for myself without interference of bureaucrats or rule-mongers. -- _____________________________ Bill http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From fabienfuente <@t> yahoo.fr Thu May 13 11:13:44 2004 From: fabienfuente <@t> yahoo.fr (Fabien FUENTE) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Phosphoprotein antibody? Message-ID: Hi, Does somebody know a human dentin phosphoprotein antibody for FACS application. If somebody have any information concerning such product, I?m very interested. Thank you very much for your help Sylvie Liabeuf From JWEEMS <@t> sjha.org Thu May 13 11:03:09 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Safety regs/Pathologists Message-ID: All well and true - unless one is caught, at which time the thousands of dollars that one may be fined should then be paid by the person not following the rules? Perhaps you don't have these strict measures with fines? It's also a real nightmare for managers who must make their staff follow the rules. j -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr, Consultant Histopathologist Sent: Thursday, May 13, 2004 11:55 AM To: Bill; histonet@pathology.swmed.edu Subject: RE: [Histonet] Safety regs/Pathologists Although Bill's views may sound rather prima donna-ish to some, I must support him. Safety procedures and practices have become rather petty, tedious, and in my view, sometimes more dangerous than the perceived danger designed for. Goggles prevent you seeing well - dangerous. Feather blade holders prevent you sensing where the blade is - dangerous. Safety hoods/cabinets prevent you seeing or doing anything in other than a clumsy way - dangerous, and so on. Let sense rule over nonsense. NB - this is NOT in any way anti-safety. 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: Bill [mailto:bill501@mindspring.com] Sent: 13 May 2004 15:19 To: histonet@pathology.swmed.edu Subject: Re: [Histonet] Safety regs/Pathologists At 9:09 AM -0400 5/13/04, Gldnsthisto@aol.com wrote: >Can anyone out there help me with what to do when the pathologists and their >staff (not paid by the hospital) will not follow safety regulations As a Pathologist and Physician, I try to never let regulations get in the way of my caring for patients. With 12 years of intensive education after high school and several decades of experience I claim, nay, demand the right to decide how to behave in a clinical setting for myself without interference of bureaucrats or rule-mongers. -- _____________________________ Bill http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ 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 thackett <@t> ils-inc.com Thu May 13 11:42:00 2004 From: thackett <@t> ils-inc.com (Theleria Hackett) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] PTAH Fixative Message-ID: Has anyone had good results on a PTAH using a fixative other than Zenker's? Thanks! Theleria R. Hackett, B.S., HT (ASCP) Histology Manager ILS, Inc. P.O. Box 13501 RTP, NC 27709 (919) 281-1110 ext. 730 From funderwood <@t> mcohio.org Thu May 13 11:43:49 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Safety regs/Pathologists Message-ID: Unfortunately, lawsuits and fines feed on nonsense and not sense. >>> "Marshall Terry Dr,Consultant Histopathologist" 05/13/04 11:55AM >>> Although Bill's views may sound rather prima donna-ish to some, I must support him. Safety procedures and practices have become rather petty, tedious, and in my view, sometimes more dangerous than the perceived danger designed for. Goggles prevent you seeing well - dangerous. Feather blade holders prevent you sensing where the blade is - dangerous. Safety hoods/cabinets prevent you seeing or doing anything in other than a clumsy way - dangerous, and so on. Let sense rule over nonsense. NB - this is NOT in any way anti-safety. 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: Bill [mailto:bill501@mindspring.com] Sent: 13 May 2004 15:19 To: histonet@pathology.swmed.edu Subject: Re: [Histonet] Safety regs/Pathologists At 9:09 AM -0400 5/13/04, Gldnsthisto@aol.com wrote: >Can anyone out there help me with what to do when the pathologists and their >staff (not paid by the hospital) will not follow safety regulations As a Pathologist and Physician, I try to never let regulations get in the way of my caring for patients. With 12 years of intensive education after high school and several decades of experience I claim, nay, demand the right to decide how to behave in a clinical setting for myself without interference of bureaucrats or rule-mongers. -- _____________________________ Bill http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ 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 pruegg <@t> colobio.com Thu May 13 12:15:10 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Phosphoprotein antibody? In-Reply-To: Message-ID: check out www.phosphosolutions.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Fabien FUENTE Sent: Thursday, May 13, 2004 10:14 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Phosphoprotein antibody? Hi, Does somebody know a human dentin phosphoprotein antibody for FACS application. If somebody have any information concerning such product, I?m very interested. Thank you very much for your help Sylvie Liabeuf _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gudrun.lang <@t> aon.at Thu May 13 12:34:07 2004 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] where's the eosin? Message-ID: <008b01c43910$8fe84380$eeeea8c0@SERVER> Dear netters We process the prostatatissue in makro-cassettes in double time of the normal schedule in a VIP. NBF-70-80-96-100-Shell.sol-Paraffin total for 24 hours. The large sections are stained H&E by hand. (self-made Mayer's haematox. 20 min; selfmade 2% Eosin with a few drops of glacial acetic acid 8 min ). Recently it happend for two times, that the tissue doesn't accept the Eosin. The sections remained blue and had only a shade of red. Normally our HE's are almost purple. Funny is, that in one case, the first slides were stained a few days before and looked good. The following slides of the same blocks remained blue without red. Could anyone give me a hint? Thanks in advance. Gudrun Lang From LESCHUKC <@t> trinity-health.org Thu May 13 12:34:21 2004 From: LESCHUKC <@t> trinity-health.org (Carmen Leschuk) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] Michigan Job Opening Message-ID: St. Joseph Mercy Oakland, in Pontiac, Michigan has a histology job opening for a full-time employee or 2 part-time employees. Please contact Carmen Leschuk at (248)858-3190 for further inquiries. Thank you! From rjr6 <@t> psu.edu Thu May 13 12:56:12 2004 From: rjr6 <@t> psu.edu (Roberta Horner) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] where's the eosin? In-Reply-To: <008b01c43910$8fe84380$eeeea8c0@SERVER> Message-ID: <003b01c43913$954dc400$8861ba92@padlspsu.psu.edu> Whenever this happens here it is usually the pH of the eosin that causes the change of staining. I keep my eosin between 4.80 and 4.90. I just had this happen about a month ago and it took me almost a week until we were no longer blue. Thank goodness the pathologists could still read the slides. Roberta Horner HT/HTL Animal Diagnostic Lab Penn State University -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Thursday, May 13, 2004 1:34 PM To: Histonetliste Subject: [Histonet] where's the eosin? Dear netters We process the prostatatissue in makro-cassettes in double time of the normal schedule in a VIP. NBF-70-80-96-100-Shell.sol-Paraffin total for 24 hours. The large sections are stained H&E by hand. (self-made Mayer's haematox. 20 min; selfmade 2% Eosin with a few drops of glacial acetic acid 8 min ). Recently it happend for two times, that the tissue doesn't accept the Eosin. The sections remained blue and had only a shade of red. Normally our HE's are almost purple. Funny is, that in one case, the first slides were stained a few days before and looked good. The following slides of the same blocks remained blue without red. Could anyone give me a hint? Thanks in advance. Gudrun Lang _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Ronnie_Houston <@t> bshsi.com Thu May 13 14:04:18 2004 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:22:57 2005 Subject: [Histonet] cpt codes for ISH Message-ID: <530361BF03351B4CAE5270A05D3037B503673279@bsrexms01.BSHSIR.COM> What cpt code(s) do you use for ISH for EBER, both technical and professional? Thanks Ronnie Ronnie Houston Director of Anatomic Pathology Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 (804) 287 7906 - fax ronnie_houston@bshsi.com ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone at 410-442-3250, and permanently delete the original e-mail, attachment(s), and any copies. From Cathy.Stevens <@t> HealthONEcares.com Thu May 13 13:49:02 2004 From: Cathy.Stevens <@t> HealthONEcares.com (Stevens Cathy) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] How do you clean your waterbaths? Message-ID: Just looking to see what various people use to clean the debris off their water baths while cutting. We use kimwipes. Anything better? Thanks Cathy Stevens H.T.(ASCP) BS Pathology Coordinator The Medical Center of Aurora P-303-695-2636 F-303-873-5660 cathy.stevens@healthonecares.com This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have recieved this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have recieve this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed. From Stacy_McLaughlin <@t> cooley-dickinson.org Thu May 13 14:32:15 2004 From: Stacy_McLaughlin <@t> cooley-dickinson.org (Stacy McLaughlin) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] How do you clean your waterbaths? Message-ID: <3D502BBF5356D31184650090275B750D0346C783@mail.cooley-dickinson.org> We use Kimwipes too, except for one tech. She prefers the yellow pages. -----Original Message----- From: Stevens Cathy [mailto:Cathy.Stevens@HealthONEcares.com] Sent: Thursday, May 13, 2004 1:49 PM To: 'HISTONET@PATHOLOGY.SWMED.EDU' Subject: [Histonet] How do you clean your waterbaths? Just looking to see what various people use to clean the debris off their water baths while cutting. We use kimwipes. Anything better? Thanks Cathy Stevens H.T.(ASCP) BS Pathology Coordinator The Medical Center of Aurora P-303-695-2636 F-303-873-5660 cathy.stevens@healthonecares.com This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have recieved this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have recieve this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet THIS E-MAIL MESSAGE 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 the reader of this e-mail message is not the intended recipient, or the employee or agent responsible for delivering the message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this e-mail message is strictly prohibited. If you have received this e-mail message in error, please immediately notify Cooley Dickinson Healthcare at 413-582-2000 and delete or shred the original message and all copies thereof. Thank you. From dlcowie <@t> prodigy.net Thu May 13 14:58:57 2004 From: dlcowie <@t> prodigy.net (Dawn Cowie) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Position open in Pensacola, Fl Message-ID: <20040513195857.82495.qmail@web81003.mail.yahoo.com> hi histonetters, I have a position open for a histotech. Minimum requirements, ASCP HT. Prefer experience with all aspects including IHC. We are a private lab located on campus of a large hospital. Approx volume, 40,000 cases per year. Located in Pensacola, Florida. If interested please call me or fax me your resume. Thanks, Dawn Cowie, Histology Supv. Pensacola Pathologists, PA 850-416-7251 phone, 850-416-4471 fax, dlcowie@prodigy.net e.mail From ALAbeyta <@t> salud.unm.edu Thu May 13 15:01:09 2004 From: ALAbeyta <@t> salud.unm.edu (Antonia Abeyta) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] microglia Message-ID: Hi all, Has anyone ever stained for microglia on frozen tissue? Our brain tissue is perfused with saline and I have tried fixing with 4% paraformaldehyde and PLP. My results for both were similar, I am seeing some microglia, but the morphology is not like it should be, and contrast isn't very good. I'm also having to use a very concentrated diltution (1:250) of an antibody that has worked at 1:1000 for some of my collegues. The difference is they are perfusing with PLP and using paraffin rather than frozen sections. Since we are using the sections for other experiments we are stuck perfusing w/saline and frozen sections...I'm about out of ideas and could use any suggestions! I am trying to get this to work for both fluorescence and ABC/DAB. Thanks, Antonia Antonia L. Abeyta Health Sciences Tech. III Community Environmental Health Program University of New Mexico Surge Bldg. Room 140 Albuquerque, NM 87131 (505) 272-4028 From cgfields <@t> lexhealth.org Thu May 13 15:08:49 2004 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] How do you clean your waterbaths? Message-ID: I have used 1/2 paper towels, phonebook pages, and kimwipes. It was a matter of cost more than anything. CFields Lex Med Ctn W.Columbia, SC > -----Original Message----- > From: Stevens Cathy [SMTP:Cathy.Stevens@HealthONEcares.com] > Sent: Thursday, May 13, 2004 2:49 PM > To: 'HISTONET@PATHOLOGY.SWMED.EDU' > Subject: [Histonet] How do you clean your waterbaths? > > Just looking to see what various people use to clean the debris off their > water baths while cutting. We use kimwipes. Anything better? > Thanks > > Cathy Stevens H.T.(ASCP) BS > Pathology Coordinator > The Medical Center of Aurora > P-303-695-2636 > F-303-873-5660 > cathy.stevens@healthonecares.com > > This email and any files transmitted with it may contain PRIVILEGED or > CONFIDENTIAL information and may be read or used only by the intended > recipient. If you are not the intended recipient of the email or any of > its > attachments, please be advised that you have recieved this email in error > and that any use, dissemination, distribution, forwarding, printing, or > copying of this email or any attached files is strictly prohibited. If you > have recieve this email in error, please immediately purge it and all > attachments and notify the sender by reply email or contact the sender at > the number listed. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From richard.rodriguez <@t> spcorp.com Thu May 13 15:17:58 2004 From: richard.rodriguez <@t> spcorp.com (Rodriguez, Richard) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Mega-cassette colors Message-ID: <4508920F80C0D411B90200508BF9A9F4049324AC@LAFMSG30.us.schp.com> I'm looking to find vendors that manufacture mega-cassettes in colors other than white. I've had some samples in the past but do not know where they came from. Please let me know if anyone knows of any out there. Thanks Richard Rodriguez Scientist 1 Schering-Plough Research Institute 144 Rt. 94 P.O. Box 32 Lafayette, NJ 07848 973-940-4282 fax: 973-940-4120 ********************************************************************* 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 naje1972 <@t> yahoo.com Thu May 13 15:30:55 2004 From: naje1972 <@t> yahoo.com (cynthia haynes) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] mega cassettes Message-ID: <20040513203055.56980.qmail@web41709.mail.yahoo.com> If anyone finds out what company is selling the colored mega cassettes please let me know also. I can be reached at naje1972@yahoo.com Thanks in advance. Cynthia Haynes H.T. __________________________________ Do you Yahoo!? Yahoo! Movies - Buy advance tickets for 'Shrek 2' http://movies.yahoo.com/showtimes/movie?mid=1808405861 From shive003 <@t> umn.edu Thu May 13 15:45:44 2004 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Fw: IHC Vet Stain Message-ID: <011001c4392b$42d63cd0$78065486@vdl220FAC> A request from a colleague who's not a subscriber. Please respond directly to LAURA at her marshfield email address below: ----- Original Message ----- From: To: Sent: Thursday, May 13, 2004 1:16 PM Subject: IHC Vet Stain > > Hey Jan, > > Do you do (or know of anyone who does) Avian Herpes testing by IHC on paraffin sections? They specifically are asking for Avian. > > Thanks much, > Laura Bliven > Marshfield Laboratories > bliven.laura@marshfieldclinic.org > > From mcauliff <@t> umdnj.edu Thu May 13 18:50:05 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] microglia In-Reply-To: References: Message-ID: <40A409AD.5060501@umdnj.edu> Hi Antonia: What animal and what antibody are you using? Mac1 or F4/80 work fine on frozen sections of mouse CNS. For rat CNS, OX-42 works on frozen sections. Lots of literature on these. Can you get an animal to fix by perfusion so you could compare your results on saline-fresh frozen versus perfused? Geoff Antonia Abeyta wrote: >Hi all, > >Has anyone ever stained for microglia on frozen tissue? Our brain >tissue is perfused with saline and I have tried fixing with 4% >paraformaldehyde and PLP. My results for both were similar, I am seeing >some microglia, but the morphology is not like it should be, and >contrast isn't very good. I'm also having to use a very concentrated >diltution (1:250) of an antibody that has worked at 1:1000 for some of >my collegues. The difference is they are perfusing with PLP and using >paraffin rather than frozen sections. Since we are using the sections >for other experiments we are stuck perfusing w/saline and frozen >sections...I'm about out of ideas and could use any suggestions! I am >trying to get this to work for both fluorescence and ABC/DAB. > >Thanks, >Antonia > >Antonia L. Abeyta >Health Sciences Tech. III >Community Environmental Health Program >University of New Mexico >Surge Bldg. Room 140 >Albuquerque, NM 87131 >(505) 272-4028 > >_______________________________________________ >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 peptolab <@t> hamptons.com Thu May 13 15:43:20 2004 From: peptolab <@t> hamptons.com (peptolab) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Automated stainer Message-ID: <002101c4392a$ee2b95e0$687bbd18@JEFF> My old Leica workhorse just broke down. Who likes or dislikes the Sakura or Leica XL autostainers. Jeff Silverman From christinmyhart <@t> yahoo.com Thu May 13 16:50:07 2004 From: christinmyhart <@t> yahoo.com (Venita Capaldo) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] (no subject) Message-ID: <20040513215007.49912.qmail@web12605.mail.yahoo.com> Hi there all. My lab is looking into purchasing a new processor and I would love some input. We are considering the Shandon Excelsior, the Tissue-Tek VIP5 and the Leica TP1050. If anyone has any input on these processors It would be greatly appreciated. We are a small dermatopathology lab and we routinly process 200-300 specimens per week. All of the specimens are skin or soft tissue and most are only 1mm to 5mm in size, but we do do some larger specimens as well. Thanks in advance for your thoughts! Sincerely, Venita Capaldo Christinmyhart@yahoo.com Allergy and Dermatology Specialists, Inc. Phoenix, Arizona (602)277-7686 --------------------------------- Do you Yahoo!? Yahoo! Movies - Buy advance tickets for 'Shrek 2' From scoop <@t> mail.nih.gov Thu May 13 17:08:01 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] cox and sdh staining Message-ID: Dear Histonetters, I would like to stain for cytochrome oxidase and succinate dehydrogenase in the same or serial sections from the same animal per the protocols in John Kiernan's book. The COX protocol says to briefly perfuse to get rid of blood and fix in formaldehyde for 15 secs and then sucrose. The SDH protocol says to use fresh frozen tissue. Can I use fresh frozen tissue for the COX protocol and quickly immersion fix in formaldehyde? Also, could people direct me to a good source of high quality formaldehyde or can I use paraformaldehyde to fix and at what concentration? I currently use paraformaldehyde from EMS. Thanks in advance for answering my (probably very ignorant, I'm not a histologist) questions. Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From convmcm <@t> cc.usu.edu Thu May 13 17:05:32 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] where's the eosin? In-Reply-To: <008b01c43910$8fe84380$eeeea8c0@SERVER> Message-ID: <002d01c43936$69a5d770$4a737b81@Cygnus> This reminds me of a funny thing that happened here some time ago. We had a graduate student (studied toxicology) working for a summer while he awaited Fall semester to begin. I trained him to do all the solutions preps and embedding, etc. He did extremely well at everything I taught him. One day, the H&Es were all H and no E. I thought something was wrong with the autostainer, so I took them back to 95%, back to eosin for 2 minutes and still no E. I smelled the eosin and guess what... no acetic acid. I should have noticed the color, but it just didn't register at the time. I asked the grad student if he added acetic acid to the eosin. He said no because it was such a small amount he didn't think it mattered (I think the acetic acid bottle was empty ==> some effort to go fill it up). I nearly died. I bit my lips till they bled that day. Anyway, acetic acid is the first thing I check with missing eosin. Hope this helps. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Thursday, May 13, 2004 10:34 AM To: Histonetliste Subject: [Histonet] where's the eosin? Dear netters We process the prostatatissue in makro-cassettes in double time of the normal schedule in a VIP. NBF-70-80-96-100-Shell.sol-Paraffin total for 24 hours. The large sections are stained H&E by hand. (self-made Mayer's haematox. 20 min; selfmade 2% Eosin with a few drops of glacial acetic acid 8 min ). Recently it happend for two times, that the tissue doesn't accept the Eosin. The sections remained blue and had only a shade of red. Normally our HE's are almost purple. Funny is, that in one case, the first slides were stained a few days before and looked good. The following slides of the same blocks remained blue without red. Could anyone give me a hint? Thanks in advance. Gudrun Lang _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Thu May 13 18:08:42 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] microwave processing Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E1BE@simba.kids> Dawn. For small or thin biopsies, you probably don't need the vacuum. You are probably over processing the tissues. Try shortening the time/dropping the temp a few degrees. The vacuum attachment is brilliant for thicker blocks. Could you post some pics to the Histonet Pics site so we could have a look? "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." Hope this helps, 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 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html -----Original Message----- From: Dawn Cowie [mailto:dlcowie@prodigy.net] Sent: Thursday, 13 May 2004 4:42 AM To: histonet Subject: [Histonet] microwave processing hi histonetters, Is anyone using the RHS-1 microwave tissue processor with vacuum? If so, would you care to share with me the processing schedule you use for tiny bx's such as gastric, liver cores, etc. We have not been having great success so far. Any help would be appreciated. Thanks, Dawn Cowie HT _______________________________________________ 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 AnthonyH <@t> chw.edu.au Thu May 13 18:21:14 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Safety regs/Pathologists Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E1BF@simba.kids> Bill, I assume you also have the welfare of your staff at heart when you embark on your clinical duties. Otherwise if people around you are AT RISK then be ready to be sued and if a death occurs you could face a jail term. Sobering? We must be forever vigilant. 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 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html -----Original Message----- From: Bill [mailto:bill501@mindspring.com] Sent: Friday, 14 May 2004 12:19 AM To: histonet@pathology.swmed.edu Subject: Re: [Histonet] Safety regs/Pathologists At 9:09 AM -0400 5/13/04, Gldnsthisto@aol.com wrote: >Can anyone out there help me with what to do when the pathologists and their >staff (not paid by the hospital) will not follow safety regulations As a Pathologist and Physician, I try to never let regulations get in the way of my caring for patients. With 12 years of intensive education after high school and several decades of experience I claim, nay, demand the right to decide how to behave in a clinical setting for myself without interference of bureaucrats or rule-mongers. -- _____________________________ Bill http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ 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 AnthonyH <@t> chw.edu.au Thu May 13 18:33:34 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Safety regs/Pathologists Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E1C0@simba.kids> Terry, I must respond! Goggles prevent you seeing well - dangerous. - A splash of formalin in the eye hurts like hell - get better goggles Feather blade holders prevent you sensing where the blade is - dangerous. - I can't see how you can't sense where the blade is - It right before your eyes! - what does this mean? Safety hoods/cabinets prevent you seeing or doing anything in other than a clumsy way - dangerous, and so on - I have never found this to be the case Let sense rule over nonsense - So far the above sited examples dont fall into the nonsense category - You will have to do better! NB - this is NOT in any way anti-safety. 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 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 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html ********************************************************************** 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 bill501 <@t> mindspring.com Thu May 13 19:10:58 2004 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Safety regs/Pathologists In-Reply-To: <1CF2E2E5BB36D5119E7A0008C791F3740800E1BF@simba.kids> References: <1CF2E2E5BB36D5119E7A0008C791F3740800E1BF@simba.kids> Message-ID: At 9:21 AM +1000 5/14/04, Tony Henwood wrote: >I assume you also have the welfare of your staff at heart when you embark on >your clinical duties. Otherwise if people around you are AT RISK then be >ready to be sued and if a death occurs you could face a jail term. Sobering? Oh Please. Common sense rules. Not regulations. I've received about 10 threats about fines, loss of privilages, jail, etc. I thinks some specifics are in order this make this discussion useful. Give me some examples of regulation violations by Pathologists that have put your lives at risk. -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board From AnthonyH <@t> chw.edu.au Thu May 13 20:13:38 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Safety regs/Pathologists Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E1C1@simba.kids> Bill, I can't give you some examples of reg violations because: 1 My pathologists (and scientific & Technical staff) aren't allowed to break Regs. 2. I can't remember any specific cases in the past. They probably were there but possibly before Regs came into force. If we did find something that was unsafe, we introduced Safe Working Practices to ensure staff safety. Can you give examples where the regs are ridiculous and are counter to common sense? Otherwise, follow the Regs and chances are you will not be injured! 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: Bill Blank [mailto:bill501@mindspring.com] Sent: Friday, 14 May 2004 10:11 AM To: histonet@pathology.swmed.edu Subject: RE: [Histonet] Safety regs/Pathologists At 9:21 AM +1000 5/14/04, Tony Henwood wrote: >I assume you also have the welfare of your staff at heart when you embark on >your clinical duties. Otherwise if people around you are AT RISK then be >ready to be sued and if a death occurs you could face a jail term. Sobering? Oh Please. Common sense rules. Not regulations. I've received about 10 threats about fines, loss of privilages, jail, etc. I thinks some specifics are in order this make this discussion useful. Give me some examples of regulation violations by Pathologists that have put your lives at risk. -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ 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 bill501 <@t> mindspring.com Thu May 13 21:07:22 2004 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Safety regs/Pathologists In-Reply-To: <1CF2E2E5BB36D5119E7A0008C791F3740800E1C1@simba.kids> References: <1CF2E2E5BB36D5119E7A0008C791F3740800E1C1@simba.kids> Message-ID: At 11:13 AM +1000 5/14/04, Tony Henwood wrote: >I can't give you some examples of reg violations because: > >1 My pathologists (and scientific & Technical staff) aren't allowed to >break Regs. This sounds like a power trip to me. At 11:13 AM +1000 5/14/04, Tony Henwood wrote: >Can you give examples where the regs are ridiculous and are counter to >common sense? Wearing floppy gloves while trying to draw blood or start an IV. They provide NO protection and make the likelihood of a fingerstick greater. Refusing to give mouth to mouth cause one has not a mouthmask handy Docs and hospitals refusing to give me patient info necessary to interpret a surgical or cytology because of HIPPA craziness. Telling me to throw away my Coke when I walk into the tab on my way to the supervisor's office to review QC. (that'll get you a 1 finger ciao) Telling me my shoes or dress is inappropriate. Quoting some rule when I open a smell a specimen container to see if it is really in fixative PAP smear retrospective review on myself. Telling me I cant carpet or put a rug in my office Telling me to put one way valves on sinks which are physically impossible to draw grey water into water lines. Posting OSHA regs, hand washing rules... It makes more sense to wash ones hands BEFORE going to the john in a surg path lab. Being forced to sign out a medicaid placenta within 72 hours even if it means I have to gross it raw on Thursday. -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board From AnthonyH <@t> chw.edu.au Thu May 13 21:51:10 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Safety regs/Pathologists Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E1C3@simba.kids> Bill, At 11:13 AM +1000 5/14/04, Tony Henwood wrote: >I can't give you some examples of reg violations because: > >1 My pathologists (and scientific & Technical staff) aren't allowed to >break Regs. This sounds like a power trip to me. Is it? Or is it just that we take pride in protecting ourselves and our co-workers! At 11:13 AM +1000 5/14/04, Tony Henwood wrote: >Can you give examples where the regs are ridiculous and are counter to >common sense? Wearing floppy gloves while trying to draw blood or start an IV. They provide NO protection and make the likelihood of a fingerstick greater. Easy, wear fitted gloves - arn't floppy gloves against the Regs? Refusing to give mouth to mouth cause one has not a mouthmask handy I hope you are referring to CPR, otherwise Sexual Harrassment is more the problem! A hard call. A person might dies if you do nothing. I would leave it up to the individual. Docs and hospitals refusing to give me patient info necessary to interpret a surgical or cytology because of HIPPA craziness. Your report is therefore vague because of lack of information - we do not have this problem in Australia where a consultant is given the information in order to help the Clinician get a worthwhile report - otherwise the report states that it limited by the lack of clinical info. Telling me to throw away my Coke when I walk into the tab on my way to the supervisor's office to review QC. (that'll get you a 1 finger ciao) No food in the Lab, "I was only passing through" - keep it black and white and "I only left it on the bench while I washed my hands" will not occur. Telling me my shoes or dress is inappropriate. Why do we wear safety shoes? Why do we wear lab gowns - aren't the answers obvious? Quoting some rule when I open a smell a specimen container to see if it is really in fixative Yes, I have done this in the past - gee it hurts the nose - There are quick tests you can do - add a drop of Schiff's reagent to a sample of the fluid and watch the colour change to pink if formalin is present. PAP smear retrospective review on myself. I've never had a PAP smear. If you are referring to QA reviews, then why should it be a problem. Telling me I cant carpet or put a rug in my office. Carpet dust is bad for microscopes. Granted this reg is inappropriate for an office. Telling me to put one way valves on sinks which are physically impossible to draw grey water into water lines. Is it? - I'm not a plumber, so I will accept what the expert's tell me. Posting OSHA regs, hand washing rules... It makes more sense to wash ones hands BEFORE going to the john in a surg path lab. One of our Regs states that Hands must be washed prior to leaving the Lab - so this should not be a problem. Being forced to sign out a medicaid placenta within 72 hours even if it means I have to gross it raw on Thursday. In Australia, we would not do this, our recommendations recommend that 80% of reports on such specimens be available within 3 days. I would like to see this changed to working days but there you are. 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 _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board ********************************************************************** 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 mab70 <@t> medschl.cam.ac.uk Fri May 14 02:29:09 2004 From: mab70 <@t> medschl.cam.ac.uk (Margaret Blount) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Histology bench ergonomics Message-ID: <6C21D947F7189448AB447C57BF2992B4025F16B9@mius.medlan.cam.ac.uk> For what it is worth, I have worked at both heights and prefer desk (30") height - it's more comfortable. After all comfort is part of ergonomics. Margaret Margaret Blount Chief Technician Clinical Biochemistry University of Cambridge Addenbrooke's Hospital Hills Road Cambridge CB2 2QR -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jackie.O'Connor@abbott.com Sent: Thursday, May 13, 2004 4:12 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Histology bench ergonomics Does anyone have any data or information on bench height for microtomy? I know a lot of people cut at routine lab bench height (36" I think) with no trouble, and some cut at desk high benches (30"). Is there any safety/ergonomic guidelines or opinions as to which is better for microtomy? I actually prefer desk height, so I can keep my feet on the floor - I'm just wondering if there is anything official out there - I've looked at a couple of sites, but can't find a lot to support my idea that desk height is better. Thanks - Jackie O' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mab70 <@t> medschl.cam.ac.uk Fri May 14 02:36:50 2004 From: mab70 <@t> medschl.cam.ac.uk (Margaret Blount) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] (no subject) Message-ID: <6C21D947F7189448AB447C57BF2992B4025F16BA@mius.medlan.cam.ac.uk> I have a Leica TP1020, it is fine but I would suggest you get a third wax bath with it or a separate vacuum infiltration system if they are still on the market. Margaret Margaret Blount Chief Technician Clinical Biochemistry University of Cambridge Addenbrooke's Hospital Hills Road Cambridge CB2 2QR -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Venita Capaldo Sent: Thursday, May 13, 2004 10:50 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Hi there all. My lab is looking into purchasing a new processor and I would love some input. We are considering the Shandon Excelsior, the Tissue-Tek VIP5 and the Leica TP1050. If anyone has any input on these processors It would be greatly appreciated. We are a small dermatopathology lab and we routinly process 200-300 specimens per week. All of the specimens are skin or soft tissue and most are only 1mm to 5mm in size, but we do do some larger specimens as well. Thanks in advance for your thoughts! Sincerely, Venita Capaldo Christinmyhart@yahoo.com Allergy and Dermatology Specialists, Inc. Phoenix, Arizona (602)277-7686 --------------------------------- Do you Yahoo!? Yahoo! Movies - Buy advance tickets for 'Shrek 2' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hcha <@t> epitomics.com Fri May 14 03:40:05 2004 From: hcha <@t> epitomics.com (Helen Cha) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Phosphoprotein antibody? Message-ID: <2E75ACE4B7DE864AA05C94B5FB6032981BA50F@epidom.epitomicsoffice.com> Hi Sylvie, We have the only rabbit monoclonal antibody technology available and we've been very successful at generating phospho-specific antibodies. Unfortunately we only offer custom service at this time, but I will let our in-house development team know so we might offer it in the near future. Thanks, Helen Helen H. Cha, Ph.D. Epitomics, Inc. www.epitomics.com Tel: (650) 583-6688 Ext. 265 E-mail: hcha@epitomics.com Subject: [Histonet] Phosphoprotein antibody? Hi, Does somebody know a human dentin phosphoprotein antibody for FACS application. If somebody have any information concerning such product, I'm very interested. Thank you very much for your help Sylvie Liabeuf From ree3 <@t> leicester.ac.uk Fri May 14 04:37:14 2004 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Safety regs/Pathologists Message-ID: The trouble with the arrogant old fart(AOFs) approach to Health and Safety matters is not that the AOFs put anyone or themselves in any danger as they have that most valuable of commodities, experience, but this is just what their impressionable juniors(IRs) do not have and it is these IRs that will go on and make the mistakes. Richard Edwards(aged 56 and a quarter) MRC TOX UNIT LEICESTER..U.K.. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 13 May 2004 16:55 To: Bill; histonet@pathology.swmed.edu Subject: RE: [Histonet] Safety regs/Pathologists Although Bill's views may sound rather prima donna-ish to some, I must support him. Safety procedures and practices have become rather petty, tedious, and in my view, sometimes more dangerous than the perceived danger designed for. Goggles prevent you seeing well - dangerous. Feather blade holders prevent you sensing where the blade is - dangerous. Safety hoods/cabinets prevent you seeing or doing anything in other than a clumsy way - dangerous, and so on. Let sense rule over nonsense. NB - this is NOT in any way anti-safety. 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: Bill [mailto:bill501@mindspring.com] Sent: 13 May 2004 15:19 To: histonet@pathology.swmed.edu Subject: Re: [Histonet] Safety regs/Pathologists At 9:09 AM -0400 5/13/04, Gldnsthisto@aol.com wrote: >Can anyone out there help me with what to do when the pathologists and their >staff (not paid by the hospital) will not follow safety regulations As a Pathologist and Physician, I try to never let regulations get in the way of my caring for patients. With 12 years of intensive education after high school and several decades of experience I claim, nay, demand the right to decide how to behave in a clinical setting for myself without interference of bureaucrats or rule-mongers. -- _____________________________ Bill http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ 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 t-sherman <@t> comcast.net Fri May 14 05:13:17 2004 From: t-sherman <@t> comcast.net (Todd Sherman) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Re: cox and sdh staining Message-ID: <40A49BBD.30506@comcast.net> -----BEGIN PGP SIGNED MESSAGE----- Hash: SHA1 Hi Sharon, Just to clear up any confusion for others, COX can also stand for CycloOXegenase instead of cytochrome oxidase. I've seen cytochrome oxidase referred to as CO and CcO. Sorry I cannot address your specific issues. Todd Todd Sherman President HistoSoft Corporation "Biology in a new form..." Home: www.histosoft.com Member Services: www.myhistosoft.com histonet-request@lists.utsouthwestern.edu wrote: | Today's Topics: | | 17. cox and sdh staining (Sharon Cooperman) | ---------------------------------------------------------------------- Message: 17 Date: Thu, 13 May 2004 18:08:01 -0400 From: Sharon Cooperman Subject: [Histonet] cox and sdh staining To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" Dear Histonetters, I would like to stain for cytochrome oxidase and succinate dehydrogenase in the same or serial sections from the same animal per the protocols in John Kiernan's book. The COX protocol says to briefly perfuse to get rid of blood and fix in formaldehyde for 15 secs and then sucrose. The SDH protocol says to use fresh frozen tissue. Can I use fresh frozen tissue for the COX protocol and quickly immersion fix in formaldehyde? Also, could people direct me to a good source of high quality formaldehyde or can I use paraformaldehyde to fix and at what concentration? I currently use paraformaldehyde from EMS. Thanks in advance for answering my (probably very ignorant, I'm not a histologist) questions. Sharon - -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 | ---------------------------------------------------------------------- -----BEGIN PGP SIGNATURE----- Version: GnuPG v1.2.3 (MingW32) Comment: Using GnuPG with MultiZilla - http://enigmail.mozdev.org iD8DBQFApJu1EmHXdrslGRcRAogMAKCOQAkjl3keAraA/RGJnWxfc4ftGACfXovv CYWKW+EkNBOf9SP22VhvdC0= =/DIu -----END PGP SIGNATURE----- From ESerrano <@t> imim.es Fri May 14 05:24:12 2004 From: ESerrano <@t> imim.es (Serrano del Pozo, Erika) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] DAB and beta-galactosidase Message-ID: <10708933FD0CD511BCFE000102BE5F72025FAD1F@hpserv.imim.es> Hello Anne, hi histonet list, I have a problem similar the one you had a couple of years ago in reference to the DAB and the beta-galactosidase. I am using in rosa26 mice tissue (that express beta-galactosidase) the policlonal antibody of Cortex Biochem, and the negative controls, as it happened to you gives me a positive stain when revealing with DAB... , without primary antibody in a possitive tissue and with antibody in a negative tissue. Were you able to solve your problem? And does somebody know if this antibody makes positive stain in nucleus or in cytoplasm (the DAB stain I was talking about gives me possitive in nucleous, but I have seen articles where common X-gal staining is positive in cytoplasm...) Can anybody help me, please?!?!? Thanks in advance. Erika Serrano Centre de Regulaci? Gen?mica Differenciation and Cancer Programme Barcelona -SPAIN- From t-sherman <@t> comcast.net Fri May 14 05:55:11 2004 From: t-sherman <@t> comcast.net (Todd Sherman) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Re: Safety regs/Pathologists - brief rant Message-ID: <40A4A58F.5010603@comcast.net> -----BEGIN PGP SIGNED MESSAGE----- Hash: SHA1 Bill, I'm sorry... DR. Blank. I'm sure you're a very knowledeable physician, but I'm wondering who is really on the "power trip" here? The regulations are posted for everyone in the hopes of creating the safest environment possible, again, for everyone. You seem to think that the rules shouldn't apply to you because of your vast experience and "demand the right to decide...without interference..." If you are dissatisfied with the industry regulations or hospital laboratory policy, is there not a committee or forum to which you can air your complaints? Maybe you could get together with colleagues that share your views and form your own advisory panel. Gosh, with all of your experience, I'm sure it would be a humdinger. I think you owe Mr. Henwood an apology. Todd Sherman - -----Original Message----- From: Bill [mailto:bill501@mindspring.com] Sent: Friday, 14 May 2004 12:19 AM To: histonet@pathology.swmed.edu Subject: Re: [Histonet] Safety regs/Pathologists At 9:09 AM -0400 5/13/04, Gldnsthisto@aol.com wrote: |>Can anyone out there help me with what to do when the pathologists and their |>staff (not paid by the hospital) will not follow safety regulations As a Pathologist and Physician, I try to never let regulations get in the way of my caring for patients. With 12 years of intensive education after high school and several decades of experience I claim, nay, demand the right to decide how to behave in a clinical setting for myself without interference of bureaucrats or rule-mongers. Bill Message: 24 Date: Thu, 13 May 2004 21:07:22 -0500 From: Bill Blank Subject: RE: [Histonet] Safety regs/Pathologists To: histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" At 11:13 AM +1000 5/14/04, Tony Henwood wrote: |>I can't give you some examples of reg violations because: |> |>1 My pathologists (and scientific & Technical staff) aren't allowed to |>break Regs. This sounds like a power trip to me. histonet-request@lists.utsouthwestern.edu wrote: | Today's Topics: | | 24. RE: Safety regs/Pathologists (Bill Blank) | | ---------------------------------------------------------------------- -----BEGIN PGP SIGNATURE----- Version: GnuPG v1.2.3 (MingW32) Comment: Using GnuPG with MultiZilla - http://enigmail.mozdev.org iD8DBQFApKWLEmHXdrslGRcRAl1gAJ4oHQiDnFwQs0ZBdu1XitTCwdhtUQCgnsK7 hytE4WMpvfg7L9qubrgDhK8= =jIL5 -----END PGP SIGNATURE----- From MDiCarlo <@t> KaleidaHealth.Org Fri May 14 06:29:40 2004 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] mega cassettes Message-ID: I received a sample mega cassette from Surgipath but it is white. Maybe they have colored ones too. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 -----Original Message----- From: cynthia haynes [mailto:naje1972@yahoo.com] Sent: Thursday, May 13, 2004 16:31 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] mega cassettes If anyone finds out what company is selling the colored mega cassettes please let me know also. I can be reached at naje1972@yahoo.com Thanks in advance. Cynthia Haynes H.T. __________________________________ Do you Yahoo!? Yahoo! Movies - Buy advance tickets for 'Shrek 2' http://movies.yahoo.com/showtimes/movie?mid=1808405861 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to 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, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. From funderwood <@t> mcohio.org Fri May 14 07:09:44 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] mega cassettes Message-ID: Surgipath does have colored super cassettes(blue, orange and gray) and they have the base molds. It looks like you could bake a cake in them. Thermo-Shandon has green mega cassettes. Fred >>> "DiCarlo, Margaret" 05/14/04 07:29AM >>> I received a sample mega cassette from Surgipath but it is white. Maybe they have colored ones too. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 -----Original Message----- From: cynthia haynes [mailto:naje1972@yahoo.com] Sent: Thursday, May 13, 2004 16:31 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] mega cassettes If anyone finds out what company is selling the colored mega cassettes please let me know also. I can be reached at naje1972@yahoo.com Thanks in advance. Cynthia Haynes H.T. __________________________________ Do you Yahoo!? Yahoo! Movies - Buy advance tickets for 'Shrek 2' http://movies.yahoo.com/showtimes/movie?mid=1808405861 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to 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, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JSCHUMA1 <@t> FAIRVIEW.ORG Fri May 14 07:50:39 2004 From: JSCHUMA1 <@t> FAIRVIEW.ORG (JENNIFER SCHUMACHER) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Embedding centers Message-ID: We need to buy a new embedding center (to replace very old Tissue Tek units). We are being asked to make a decision without the luxury of any demo units. I would welcome any suggestions/ideas . . . also WHY you feel a particular unit is the best. Thanks in advance. Happy Friday!! Jennifer 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 brett_connolly <@t> merck.com Fri May 14 08:37:18 2004 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Mouse on mouse kits Message-ID: Has anyone evaluated the various MOM kits side by side? I'm looking for recommendations for those that provide the cleanest background. Thx, Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP26A-3000 PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-652-2075 e-mail. brett_connolly@merck.com ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ From Terry.Marshall <@t> rothgen.nhs.uk Fri May 14 08:07:15 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Safety regs/Pathologists (males only) Message-ID: Bill says: "It makes more sense to wash ones hands BEFORE going to the john in a surg path lab." Absolutely. The "Now wash your hands" sign in a urinal has always seemed plain wrong to me. I get up and shower. I put on clean underpants. My penis is spotless. I go to work. After a couple of hours, I go get a pee and put my filthy hands over my nice clean penis. Now I'm told to wash my hands. Crazy - I should wash my penis. 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: Bill Blank [mailto:bill501@mindspring.com] Sent: 14 May 2004 03:07 To: histonet@pathology.swmed.edu Subject: RE: [Histonet] Safety regs/Pathologists At 11:13 AM +1000 5/14/04, Tony Henwood wrote: >I can't give you some examples of reg violations because: > >1 My pathologists (and scientific & Technical staff) aren't allowed to >break Regs. This sounds like a power trip to me. At 11:13 AM +1000 5/14/04, Tony Henwood wrote: >Can you give examples where the regs are ridiculous and are counter to >common sense? Wearing floppy gloves while trying to draw blood or start an IV. They provide NO protection and make the likelihood of a fingerstick greater. Refusing to give mouth to mouth cause one has not a mouthmask handy Docs and hospitals refusing to give me patient info necessary to interpret a surgical or cytology because of HIPPA craziness. Telling me to throw away my Coke when I walk into the tab on my way to the supervisor's office to review QC. (that'll get you a 1 finger ciao) Telling me my shoes or dress is inappropriate. Quoting some rule when I open a smell a specimen container to see if it is really in fixative PAP smear retrospective review on myself. Telling me I cant carpet or put a rug in my office Telling me to put one way valves on sinks which are physically impossible to draw grey water into water lines. Posting OSHA regs, hand washing rules... It makes more sense to wash ones hands BEFORE going to the john in a surg path lab. Being forced to sign out a medicaid placenta within 72 hours even if it means I have to gross it raw on Thursday. -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Fri May 14 08:38:33 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Automated stainer Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB3FC6@fh2k093.fhmis.net> We use the Leica1050 processor and the XL stainers - both excellent workhorses. our VIP gets plugged allot and the selenoids need replacing under the same conditions. The Leicas have been here longer than I have (6 years) and the VIPs are newer than I am. -----Original Message----- From: peptolab [mailto:peptolab@hamptons.com] Sent: Thursday, May 13, 2004 4:43 PM To: HistoNet Server Subject: [Histonet] Automated stainer My old Leica workhorse just broke down. Who likes or dislikes the Sakura or Leica XL autostainers. Jeff Silverman _______________________________________________ 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 LESCHUKC <@t> trinity-health.org Fri May 14 09:07:09 2004 From: LESCHUKC <@t> trinity-health.org (Carmen Leschuk) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Automated stainer Message-ID: I would definitely recommend the Tissue-Tek VIP. It is the best processor out there by far. >>> "Bonner, Janet" 05/14/04 09:38AM >>> We use the Leica1050 processor and the XL stainers - both excellent workhorses. our VIP gets plugged allot and the selenoids need replacing under the same conditions. The Leicas have been here longer than I have (6 years) and the VIPs are newer than I am. -----Original Message----- From: peptolab [mailto:peptolab@hamptons.com] Sent: Thursday, May 13, 2004 4:43 PM To: HistoNet Server Subject: [Histonet] Automated stainer My old Leica workhorse just broke down. Who likes or dislikes the Sakura or Leica XL autostainers. Jeff Silverman _______________________________________________ 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 From adelsyscarol <@t> yahoo.com Fri May 14 09:31:04 2004 From: adelsyscarol <@t> yahoo.com (Carol Wilson) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Looking to buy tissue-tek dispensing console Message-ID: <20040514143104.17267.qmail@web50709.mail.yahoo.com> Looking to purchase a tissue-tek dispensing console in working condition. Please contact me directly at Adelsyscarol@yahoo.com Thank you. Carol Wilson Adelysis, Inc (216)932-7500 This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. --------------------------------- Do you Yahoo!? SBC Yahoo! - Internet access at a great low price. From ian.montgomery <@t> bio.gla.ac.uk Fri May 14 09:34:10 2004 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:22:58 2005 Subject: Fwd: [Histonet] Mouse on mouse kits Message-ID: <6.1.0.6.2.20040514153234.02d122b0@udcf.gla.ac.uk> Brett, My only experience is with Dako Ark and so far it has been superb. Ian >Has anyone evaluated the various MOM kits side by side? I'm looking for >recommendations for those that provide the cleanest background. > >Thx, >Brett > >Brett M. Connolly, Ph.D. >Merck & Co., Inc. >MRL, Imaging Research >WP26A-3000 >PO Box 4 >West Point, PA 19486 >PH 215-652-2501 >fax. 215-652-2075 >e-mail. brett_connolly@merck.com 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 mbryhan <@t> NORTHERNHEALTH.ORG Fri May 14 10:18:36 2004 From: mbryhan <@t> NORTHERNHEALTH.ORG (Mary Bryhan) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] How do you clean your waterbaths? Message-ID: Gauze around the edges and kimwipes for the surface. Mary Bryhan >>> Stevens Cathy 05/13/04 02:49PM >>> Just looking to see what various people use to clean the debris off their water baths while cutting. We use kimwipes. Anything better? Thanks Cathy Stevens H.T.(ASCP) BS Pathology Coordinator The Medical Center of Aurora P-303-695-2636 F-303-873-5660 cathy.stevens@healthonecares.com This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have recieved this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have receive this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Fri May 14 09:54:03 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Automated stainer Message-ID: I love my sakura. Quite user friendly, versatile, snd trouble free (insert sound of knocking on wood). I've used two different models, currently have the DRS 2000. Fred >>> "peptolab" 05/13/04 04:43PM >>> My old Leica workhorse just broke down. Who likes or dislikes the Sakura or Leica XL autostainers. Jeff Silverman _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bill501 <@t> mindspring.com Fri May 14 09:58:42 2004 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Safety regs/Pathologists In-Reply-To: References: Message-ID: At 9:22 AM -0400 5/14/04, Fran Lemons wrote: >Forcing a gross assistant to stand by while gross dissection is >performed without proper ventilation can cause death over time. >Don't mean to be abrasive, but you sound like you are just plain stubborn. I absolutely agree with this. Proper ventilation for formalin and xylene, among others, is essential for my and personnel safety. Proper ventilation does not interfere with patient care and is a reasonable precaution. In my lab we use industrial strength exhaust fans and wear formalin and xylene badges at the prescribed intervals. No one has ever come up over-exposed. If a grossing assistant were bothered by such fumes or even a leg with wet gangrene I would tell them to leave if necessary. The main point I have tried to make in this brouhaha is that when one has to choose between a regulation and good health care for a patient, on should always choose the latter, IMO. No rule or reg is black and white and there are always exceptions. I consider myself and my technical supervisors and staff to have enough common sense to ignore a rule or reg if necessary or if it makes no sense in a particular circumstance. I also see some cultural differences in opinions about this. Not only UK, USA and Oz, but rural vs urban USA. There is also a generational gap apparent. I know many people live by rules and regs, because they don't have to think for themselves and make sometimes difficult decisions. They allow the rules and regs to be the axe that drops. I would rather have my staff be more independent thinking and an abundance of common sense. -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board From kemlo <@t> tiscali.co.uk Fri May 14 11:12:55 2004 From: kemlo <@t> tiscali.co.uk (Kemlo) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Safety regs/Pathologists (males only) In-Reply-To: Message-ID: <40800C1000067316@mk-cpfrontend-4.mail.uk.tiscali.com> That's you all over Terry, making a mountain out of a molehill. I'm surprised your male appendage is still attached, you must have been very skillful! __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From hymclab <@t> hyhc.com Fri May 14 12:03:08 2004 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] (no subject) Message-ID: We bought a VIP5 last fall and absolutely love it. It is very user friendly and produces quality tissue blocks. I just attended the VIP advanced users training school in California and it was awesome. They really teach you a lot of good things and trouble shooting techniques that we probably will not need. We have had the old VIP for 14 years with hardly any problems. I would highly recommend the VIP5. Dawn Schneider Lead Tech Howard Young Medical Center Woodruf, WI -----Original Message----- From: Venita Capaldo [mailto:christinmyhart@yahoo.com] Sent: Thursday, May 13, 2004 4:50 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Hi there all. My lab is looking into purchasing a new processor and I would love some input. We are considering the Shandon Excelsior, the Tissue-Tek VIP5 and the Leica TP1050. If anyone has any input on these processors It would be greatly appreciated. We are a small dermatopathology lab and we routinly process 200-300 specimens per week. All of the specimens are skin or soft tissue and most are only 1mm to 5mm in size, but we do do some larger specimens as well. Thanks in advance for your thoughts! Sincerely, Venita Capaldo Christinmyhart@yahoo.com Allergy and Dermatology Specialists, Inc. Phoenix, Arizona (602)277-7686 --------------------------------- Do you Yahoo!? Yahoo! Movies - Buy advance tickets for 'Shrek 2' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Fri May 14 11:50:06 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] How do you clean your waterbaths? Message-ID: Toilet paper. Ocassionaly you may have a problem with pieces breaking off. But hey, it's cheap and fits nicely over a wooden dowel for a handy dispenser. >>> Stevens Cathy 05/13/04 02:49PM >>> Just looking to see what various people use to clean the debris off their water baths while cutting. We use kimwipes. Anything better? Thanks Cathy Stevens H.T.(ASCP) BS Pathology Coordinator The Medical Center of Aurora P-303-695-2636 F-303-873-5660 cathy.stevens@healthonecares.com This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have recieved this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have recieve this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Fri May 14 11:44:45 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Histology bench ergonomics Message-ID: The only thing I don't recommend is standing while cutting. I did this for years, sometimes 8 hours or more at a time. Talk about spaghetti legs. >>> 05/13/04 11:11AM >>> Does anyone have any data or information on bench height for microtomy? I know a lot of people cut at routine lab bench height (36" I think) with no trouble, and some cut at desk high benches (30"). Is there any safety/ergonomic guidelines or opinions as to which is better for microtomy? I actually prefer desk height, so I can keep my feet on the floor - I'm just wondering if there is anything official out there - I've looked at a couple of sites, but can't find a lot to support my idea that desk height is better. Thanks - Jackie O' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Fri May 14 12:08:29 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Histology bench ergonomics Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA206EAE2@usca0082k08.labvision.apogent.com> Fred wrote "The only thing I don't recommend is standing while cutting" Now you're in for it Fred, The Aussie's and Kiwis are going to grill you for sure! Tim Morken -----Original Message----- From: Fred Underwood [mailto:funderwood@mcohio.org] Sent: Friday, May 14, 2004 9:45 AM To: histonet@pathology.swmed.edu Subject: Re: [Histonet] Histology bench ergonomics The only thing I don't recommend is standing while cutting. I did this for years, sometimes 8 hours or more at a time. Talk about spaghetti legs. >>> 05/13/04 11:11AM >>> Does anyone have any data or information on bench height for microtomy? I know a lot of people cut at routine lab bench height (36" I think) with no trouble, and some cut at desk high benches (30"). Is there any safety/ergonomic guidelines or opinions as to which is better for microtomy? I actually prefer desk height, so I can keep my feet on the floor - I'm just wondering if there is anything official out there - I've looked at a couple of sites, but can't find a lot to support my idea that desk height is better. Thanks - Jackie O' _______________________________________________ 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 Fri May 14 12:10:45 2004 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] How do you clean your waterbaths? Message-ID: Don't you have a lint problem? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred Underwood Sent: Friday, May 14, 2004 12:50 PM To: HISTONET@PATHOLOGY.SWMED.EDU Subject: Re: [Histonet] How do you clean your waterbaths? Toilet paper. Ocassionaly you may have a problem with pieces breaking off. But hey, it's cheap and fits nicely over a wooden dowel for a handy dispenser. >>> Stevens Cathy 05/13/04 02:49PM >>> Just looking to see what various people use to clean the debris off their water baths while cutting. We use kimwipes. Anything better? Thanks Cathy Stevens H.T.(ASCP) BS Pathology Coordinator The Medical Center of Aurora P-303-695-2636 F-303-873-5660 cathy.stevens@healthonecares.com This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have recieved this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have recieve this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed. _______________________________________________ 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 Fri May 14 12:31:23 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] How do you clean your waterbaths? Message-ID: I don't find it to be a big problem. It may not be as clean as kimwipes but the cost difference makes it worthwhile. Of course, I don't deal with biopsies, so in the event I may have to recut a block, cutting thru the specimen is not a problem. >>> "Bartlett, Jeanine" 05/14/04 01:10PM >>> Don't you have a lint problem? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred Underwood Sent: Friday, May 14, 2004 12:50 PM To: HISTONET@PATHOLOGY.SWMED.EDU Subject: Re: [Histonet] How do you clean your waterbaths? Toilet paper. Ocassionaly you may have a problem with pieces breaking off. But hey, it's cheap and fits nicely over a wooden dowel for a handy dispenser. >>> Stevens Cathy 05/13/04 02:49PM >>> Just looking to see what various people use to clean the debris off their water baths while cutting. We use kimwipes. Anything better? Thanks Cathy Stevens H.T.(ASCP) BS Pathology Coordinator The Medical Center of Aurora P-303-695-2636 F-303-873-5660 cathy.stevens@healthonecares.com This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have recieved this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have recieve this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed. _______________________________________________ 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 Fri May 14 12:21:28 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Histology bench ergonomics Message-ID: Uhhh.......Did I mention that I love a rousing match of cricket and aussie rules futbol has it all over that sissy NFL stuff. Plus Victoria Bitter is the best beer I've drank this side of Guiness. G'day Mates Fred >>> "Morken, Tim - Labvision" 05/14/04 01:08PM >>> Fred wrote "The only thing I don't recommend is standing while cutting" Now you're in for it Fred, The Aussie's and Kiwis are going to grill you for sure! Tim Morken -----Original Message----- From: Fred Underwood [mailto:funderwood@mcohio.org] Sent: Friday, May 14, 2004 9:45 AM To: histonet@pathology.swmed.edu Subject: Re: [Histonet] Histology bench ergonomics The only thing I don't recommend is standing while cutting. I did this for years, sometimes 8 hours or more at a time. Talk about spaghetti legs. >>> 05/13/04 11:11AM >>> Does anyone have any data or information on bench height for microtomy? I know a lot of people cut at routine lab bench height (36" I think) with no trouble, and some cut at desk high benches (30"). Is there any safety/ergonomic guidelines or opinions as to which is better for microtomy? I actually prefer desk height, so I can keep my feet on the floor - I'm just wondering if there is anything official out there - I've looked at a couple of sites, but can't find a lot to support my idea that desk height is better. Thanks - Jackie O' _______________________________________________ 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 lance.erickson <@t> ihc.com Fri May 14 12:41:37 2004 From: lance.erickson <@t> ihc.com (Lance Erickson) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Retic nomenclature Message-ID: OK Here is a little question I have run across while revising my Retic procedure. I had named the procedure like the original paper written by Gordon & Sweets (Gordon, H. & Sweets, H. H. A simple method for the silver impregnation of reticulum American Journal of Pathology 1936). The pathologist here that reviews the procedures said she had never heard it called reticulum and wanted me to change it to reticulin. I still am in a grey area with what it should be called. The medical dictionary says that "reticulin" is the name given to the chemical substance of reticular fibers. It also says that "reticular" means relating to a reticulum. "Reticulum" means a fine network formed by cells or formed by certain structures within cells. From this I would say that naming the procedure "Gordon & Sweets Reticulum Stain" would be correct. Further could you say "This stain demonstates reticulin" or should you say "The reticular fibers in this stain demonstate the reticulin of this portion of this tissue's reticulum" This is my Friday question for you histonetters. Can anyone clear this up for me? P.S. The dictionary also said that reticular fibers are now associated with collagen and are not as distinct as once thought and are now regarded as type III collagen. Should I just change the name to "Gordon & Sweets Type III Collagen Stain"????? I realized this has hints of sarcasm but I really do want to know the correct nomenclature. Lance Erickson Anatomic Pathology Supervisor Primary Children's Medical Center 100 N Medical Dr Salt Lake City, UT 84113-1100 (801) 588-3110 fax (801) 588-3169 From Sandra.Etheridge <@t> gems8.gov.bc.ca Fri May 14 12:55:51 2004 From: Sandra.Etheridge <@t> gems8.gov.bc.ca (Etheridge, Sandra AGF:EX) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Embedding Centers Feedback Message-ID: <424FAC08A8CDFE4BADD6497AE75B5E2F0E0A8694@atlas.gov.bc.ca> Hi, Jennifer, I have worked in five different Histology labs in the past, most of which were using Tissue Tek embedding centers probably older than myself. That says something about Tissue Tek's reliability, etc. I have also worked on the new Leica model, but we didn't like the layout as much. It also only had one block warming tray, but some of the techs liked the foot pedal option. Some techs also thought the programing was confusing. I have worked on the new Tissue Tek model and think it is more versatile and ergonomic. Just my opinion. I have ordered one for my current lab as we need a second unit. Hope this is helpful. Sandra Etheridge BC Ministry of Agriculture, Food and Fisheries Animal Health Center, Histology Abbotsford, BC Canada From LonS <@t> vetmed.wsu.edu Fri May 14 13:07:21 2004 From: LonS <@t> vetmed.wsu.edu (Lon Smith) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] job opening Message-ID: <95111571D1C6B5498C64F68DD0769BB579CA70@cvm36.vetmed.wsu.edu> The Histology department at Washington State University Animal Disease Diagnostic Laboratory has a position open for HISTOLOGIC TECHNICIAN I or HISTOLOGIC TECHNICIAN II. Applications may be requested by contacting Human Resource Services, 139 French Administration Bldg, Pullman,WA 99164-1014, Ph (509) 335-4521, or by accessing website www.hrs.wsu.edu. For additional job information, contact Marlene Nunes at (509) 335-0140. From DonnaWillis <@t> texashealth.org Fri May 14 13:50:58 2004 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] HT Practical Grading Message-ID: <5C6CBCCEB04B894C8BD15B312487F7B201A330FE@ftwex01.txhealth.org> Anyone know when the pass/fail notices will be sent to those that have recently taken their practical. I have 4 applicants that are eagerly waiting for notification. Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx 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 MinHan.Tan <@t> vai.org Fri May 14 14:40:46 2004 From: MinHan.Tan <@t> vai.org (Tan, MinHan) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] DAB (DAKO and Vector) Message-ID: <74D0F0AB07F2E647A02D839ED79520F9B48745@VAIEXCH02.vai.org> Hi, Thanks all who replied to my query on pressure cookers. I have a question for those who have used the Envision + kit. Has anyone used it with the Vector DAB kit, rather than the DAB+ from DAKO? Thanks! Min-han Tan 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 Lizbeth_Kelly <@t> hgsi.com Fri May 14 15:11:42 2004 From: Lizbeth_Kelly <@t> hgsi.com (Lizbeth_Kelly@hgsi.com) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] unsuscribe Message-ID: Please unsuscribe for now. Thanks From Charlene.Henry <@t> STJUDE.ORG Fri May 14 15:34:06 2004 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Fixation and Decalcification of Bone Marrow Bxs. Message-ID: <5CB39BCA5724F349BCB748675C6CA1A231582D@SJMEMXMB02.stjude.sjcrh.local> Fellow Histotechs, I would like to get some feedback on fixatives and decalcification protocols used in your laboratories. Our pathologists, like many, want perfection; so here is my problem. We were fixing our bone marrow bxs. in 10% NBF and then decalcification for 1 hour prior to processing. Because of the facility I work at, we perform IHC and ISH on a large percentage of our bxs. With the method we were using, some of our antibodies would not work on tissue that had been decalcified and the decalcification completely destroyed the RNA making it impossible to perform ISH. About 6 months ago, we switched to fixing our bone marrows in 10% NBF saturated with EDTA (this fixes and decals at the same time) with beautiful results with our IHC and also ISH. Even though our pathologists are elated that the IHC and ISH are great, they are not completely happy with the morphology of the H&E slides. I was just wondering if anyone has any suggestions so we can accomplish both great H&E morphology and great stains with IHC and ISH. Thanks, Charlene Henry, HT QIHC Histology/IHC Section Head Department of Pathology St. Jude Children's Research Hospital 332 North Lauderdale Memphis, TN 38105 Phone: 901-495-3349 FAX: 901-495-3100 From kwittle <@t> jhmi.edu Fri May 14 15:48:35 2004 From: kwittle <@t> jhmi.edu (Karen Wittler) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Safety Regs(males only) Message-ID: Poppy cock! There are 101 Faculty pathologists, 30 House Staff, 17 Fellows here. None of them or the 900 employees of the Pathology Department have a choice in the matter. In this teaching institution our Resident's are expected to comply with Safety Regulations and learn safe laboratory practices. All of us have to follow the same set of standards. Maybe that's one of the reasons our hospital has been named the #1 Hospital in the country for the past twelve years. (rub-rub) Karen Wittler, HT(ASCP) Johns Hopkins Hospital Baltimore, MD From funderwood <@t> mcohio.org Fri May 14 14:10:31 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Histology bench ergonomics Message-ID: Whew! Is this what it's like being a presidential canidate? Waffling and sucking up to different groups. >>> "Kyle-Byrne, Carrie - Labvision" 05/14/04 02:58PM >>> Fred, The Irish are going to pummel you for referring to Guiness as 'beer'..... I made that mistake once, and only once. It's stout. Happy Friday! Ta, Carrie -----Original Message----- From: Fred Underwood [mailto:funderwood@mcohio.org] Sent: Friday, May 14, 2004 10:21 AM To: histonet@pathology.swmed.edu Subject: RE: [Histonet] Histology bench ergonomics Uhhh.......Did I mention that I love a rousing match of cricket and aussie rules futbol has it all over that sissy NFL stuff. Plus Victoria Bitter is the best beer I've drank this side of Guiness. G'day Mates Fred >>> "Morken, Tim - Labvision" 05/14/04 01:08PM >>> Fred wrote "The only thing I don't recommend is standing while cutting" Now you're in for it Fred, The Aussie's and Kiwis are going to grill you for sure! Tim Morken -----Original Message----- From: Fred Underwood [mailto:funderwood@mcohio.org] Sent: Friday, May 14, 2004 9:45 AM To: histonet@pathology.swmed.edu Subject: Re: [Histonet] Histology bench ergonomics The only thing I don't recommend is standing while cutting. I did this for years, sometimes 8 hours or more at a time. Talk about spaghetti legs. >>> 05/13/04 11:11AM >>> Does anyone have any data or information on bench height for microtomy? I know a lot of people cut at routine lab bench height (36" I think) with no trouble, and some cut at desk high benches (30"). Is there any safety/ergonomic guidelines or opinions as to which is better for microtomy? I actually prefer desk height, so I can keep my feet on the floor - I'm just wondering if there is anything official out there - I've looked at a couple of sites, but can't find a lot to support my idea that desk height is better. Thanks - Jackie O' _______________________________________________ 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 carl.hobbs <@t> kcl.ac.uk Fri May 14 17:06:37 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] re : mouse on mouse Message-ID: <001201c439ff$ba86d9e0$5e019b51@home> Have you tried std methods and found that they give you "dirty" b'grounds? If they have not ....don't waste your money. Omit primary....if you get obscuring, non- specific positivity...fair enough. Do you want to do it on frozen or pwax sections?? Always try first...( assuming you have not already...however, no info given) --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.684 / Virus Database: 446 - Release Date: 13/05/2004 From carl.hobbs <@t> kcl.ac.uk Fri May 14 17:15:15 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] re rules?regs Message-ID: <001d01c43a00$eecdeb20$5e019b51@home> "I know many people live by rules and regs, because they don't have to think for themselves and make sometimes difficult decisions. They allow the rules and regs to be the axe that drops. I would rather have my staff be more independent thinking and an abundance of common sense." I assume you have not got a qualification that absolutely conforms to your governing body's rules and regulations? Because, without those absolute rules and regulations....sorry, you wouldn't be a practising Pathologist --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.684 / Virus Database: 446 - Release Date: 13/05/2004 From meligroc <@t> zgi.com Fri May 14 16:54:31 2004 From: meligroc <@t> zgi.com (CRME (Criss Meligro)) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Holland's Fixative for IHC Message-ID: Wondering if anyone has experience with IHC on tissue fixed in Holland's Fixative. If so what type of retrieval worked best? The tissue was received from an outside source and we have no experience with this....any help would be appreciated. THANKS! From marktarango <@t> earthlink.net Fri May 14 18:03:47 2004 From: marktarango <@t> earthlink.net (Mark Adam Tarango) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] VIP Tissue Processor malfunctioning..... In-Reply-To: <200405141001.1boG491DD3NZFjX0@robin> Message-ID: <000001c43a07$b6b0d200$2702a8c0@markie> Has anyone ever had a problem w/ their VIP processor pumping water into the last paraffin bath or retort chamber? This has happened four or five times in the last three months. It seems to always happen after I change the reagents. Am I to blame? P.S. What is the water for? Is its purpose to minimize fumes? I've heard someone say it's used to pump reagents in and out....something related to the vacuum and pressure, but this doesn't make sense to me... Thanks Mark Tarango From Gervaip <@t> aol.com Fri May 14 20:06:49 2004 From: Gervaip <@t> aol.com (Gervaip@aol.com) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Safety regs/Pathologists (males only) Message-ID: <7d.4e0fd126.2dd6c729@aol.com> As the Pathology Department Safety Officer, I give yearly safety inservice. And that is exactly what I have told them, "Wash your hands before going to the bathroom!" Makes sense to me. Pearl in New Orleans From jizv66 <@t> hotmail.com Fri May 14 20:33:34 2004 From: jizv66 <@t> hotmail.com (Jorge Ivan Zapata Valencia) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Transportation of frozen sections Message-ID: Hello everybody I want to ask how can I transport frozen sections between two distant places? I am now in Japan but I have to go back to Colombia soon and finish there the staining protocols I have to do. How I can transport the slides in such a way that my samples arrive to Colombia in good condition to be stained? Must i fix them and keep them at room temperature? Any suggestion is really appreciated Thank You very much Regards, JORGE IVAN ZAPATA _________________________________________________________________ MSN 8 helps eliminate e-mail viruses. Get 2 months FREE*. http://join.msn.com/?page=features/virus From convmcm <@t> cc.usu.edu Sat May 15 00:45:39 2004 From: convmcm <@t> cc.usu.edu (convmcm) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Safety regs/Pathologists (males only) Message-ID: <40A5B0E4@webster.usu.edu> I can't reist. THIS post is PRICELESSS!!! Woooo-hoooo Terry!!! thanks for the chuckle! I know this is a very serious discussion, but the truth is... I've been laughing my head off at all of this. Now I'm going to jump into the fray and spout of my piece. OF COURSE we need to be safe. Anyone who has worked in the pre-safety era, like me, remembers how it was to work in a small, hot, unventilated room filled with open tubs of formalin sitting on heating pads. Dipping our bare fingers into xylene while coverslipping our slides and many, many, many other things. My husband and I both have some reallly wild tales to tell about how breaches in common sense & lack of written procedures caused mayhem and disaster. When I first learned histology, it was my understanding that common sense was a necessary attribute for working in a lab of any kind. But I also believe the bureaucrats who come up with these safetly rules have NO IDEA about the dynamics of living beings. What is perfectly sensible in one lab is a disaster in another and yet we're all expected to live and work by standarized rules and regs. ALL of us want to be safe. All of us want to do the very best for our clients --- whether they're human or the animals humans care for. Common sense is TRULY the heart and soul of safety. And Common Sense is a product of a good mind coupled with knowledge. Anyone -- doctor or tech -- who likes to live by the blanket rules and regs without being aware of the reality of the moment, is the most dangerous thing in the lab. I am now donning all of the appropriate personal safety apparel in preparation for the incinerator blast that is sure to come my way ... Connie McManus Utah Veterinary Diagnostics Laboratory Utah State Universtiy 950 East 1400 North Logan, UT phone 435/797-1891 fax: 465/797-2805 >===== Original Message From "Marshall Terry Dr, Consultant Histopathologist" < Terry.Marshall@rothgen.nhs.uk> ===== >Bill says: > >"It makes more sense to wash ones hands BEFORE going to the john in a surg path lab." > >Absolutely. The "Now wash your hands" sign in a urinal has always seemed plain wrong to me. > >I get up and shower. I put on clean underpants. My penis is spotless. >I go to work. >After a couple of hours, I go get a pee and put my filthy hands over my nice clean penis. >Now I'm told to wash my hands. >Crazy - I should wash my penis. > > >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: Bill Blank [mailto:bill501@mindspring.com] >Sent: 14 May 2004 03:07 >To: histonet@pathology.swmed.edu >Subject: RE: [Histonet] Safety regs/Pathologists > > >At 11:13 AM +1000 5/14/04, Tony Henwood wrote: >>I can't give you some examples of reg violations because: >> >>1 My pathologists (and scientific & Technical staff) aren't allowed to >>break Regs. > >This sounds like a power trip to me. > >At 11:13 AM +1000 5/14/04, Tony Henwood wrote: >>Can you give examples where the regs are ridiculous and are counter to >>common sense? > >Wearing floppy gloves while trying to draw blood or start an IV. They >provide NO protection and make the likelihood of a fingerstick >greater. > >Refusing to give mouth to mouth cause one has not a mouthmask handy > >Docs and hospitals refusing to give me patient info necessary to >interpret a surgical or cytology because of HIPPA craziness. > >Telling me to throw away my Coke when I walk into the tab on my way >to the supervisor's office to review QC. (that'll get you a 1 finger >ciao) > >Telling me my shoes or dress is inappropriate. > >Quoting some rule when I open a smell a specimen container to see if >it is really in fixative > >PAP smear retrospective review on myself. > >Telling me I cant carpet or put a rug in my office > >Telling me to put one way valves on sinks which are physically >impossible to draw grey water into water lines. > >Posting OSHA regs, hand washing rules... It makes more sense to wash >ones hands BEFORE going to the john in a surg path lab. > >Being forced to sign out a medicaid placenta within 72 hours even if >it means I have to gross it raw on Thursday. > >-- >_____________________________ >Bill Blank >http://kernunnos.com (Celtic studies and numismatics) >OBOD's Message board: http://www.druidry.org/board > > > > >_______________________________________________ >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 convmcm <@t> cc.usu.edu Sat May 15 00:52:12 2004 From: convmcm <@t> cc.usu.edu (convmcm) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] How do you clean your waterbaths? Message-ID: <40A5B24B@webster.usu.edu> Hey,, that's clever! putting TP on a dowel. And it IS cheaper. One lab I used to work in used facial tissues to wipe the bath with, but that was a long time ago. I still prefer the kimwipes. Connie McManus Utah Veterinary Diagnostics Laboratory >===== Original Message From Fred Underwood ===== >I don't find it to be a big problem. It may not be as clean as >kimwipes but the cost difference makes it worthwhile. Of course, I >don't deal with biopsies, so in the event I may have to recut a block, >cutting thru the specimen is not a problem. > >>> "Bartlett, Jeanine" 05/14/04 01:10PM >>> >Don't you have a lint problem? > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred >Underwood >Sent: Friday, May 14, 2004 12:50 PM >To: HISTONET@PATHOLOGY.SWMED.EDU >Subject: Re: [Histonet] How do you clean your waterbaths? > > >Toilet paper. Ocassionaly you may have a problem with pieces breaking >off. But hey, it's cheap and fits nicely over a wooden dowel for a >handy dispenser. > > > >>> Stevens Cathy 05/13/04 02:49PM >>> >Just looking to see what various people use to clean the debris off >their water baths while cutting. We use kimwipes. Anything better? >Thanks > >Cathy Stevens H.T.(ASCP) BS >Pathology Coordinator >The Medical Center of Aurora >P-303-695-2636 >F-303-873-5660 >cathy.stevens@healthonecares.com > >This email and any files transmitted with it may contain PRIVILEGED or >CONFIDENTIAL information and may be read or used only by the intended >recipient. If you are not the intended recipient of the email or any >of >its attachments, please be advised that you have recieved this email >in >error and that any use, dissemination, distribution, forwarding, >printing, or copying of this email or any attached files is strictly >prohibited. If you have recieve this email in error, please >immediately >purge it and all attachments and notify the sender by reply email or >contact the sender at the number listed. > > >_______________________________________________ >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 jizv66 <@t> hotmail.com Sat May 15 02:46:00 2004 From: jizv66 <@t> hotmail.com (Jorge Ivan Zapata Valencia) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Fixative and H2O2 for frozen Message-ID: Hello everybody a technical question: may I use the same acetone:etanol mix to fix samples different days, or do I have to prepare it feshly every day?? The same with the H2O2 PBS solution for endogenous peroxidase blocking. Is it mandatory to use fresh one? Thank You very much jorge ivan zapata _________________________________________________________________ The new MSN 8: advanced junk mail protection and 2 months FREE* http://join.msn.com/?page=features/junkmail From RFail <@t> Charleston.net Sat May 15 05:32:09 2004 From: RFail <@t> Charleston.net (Rena Fail) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Fixation and Decalcification of Bone Marrow Bxs. In-Reply-To: <5CB39BCA5724F349BCB748675C6CA1A231582D@SJMEMXMB02.stjude.sjcrh.local> Message-ID: <000901c43a67$f2da1ec0$8a11a6a5@rena> We use formic acid for decalcification. It's slow, but the morphology is preserved and the results with IHC are excellent. Rena Fail HT(ASCP)QIHC Lead Tech IHC/SS MUSC -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Henry Charlene Sent: Friday, May 14, 2004 4:34 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Fixation and Decalcification of Bone Marrow Bxs. Fellow Histotechs, I would like to get some feedback on fixatives and decalcification protocols used in your laboratories. Our pathologists, like many, want perfection; so here is my problem. We were fixing our bone marrow bxs. in 10% NBF and then decalcification for 1 hour prior to processing. Because of the facility I work at, we perform IHC and ISH on a large percentage of our bxs. With the method we were using, some of our antibodies would not work on tissue that had been decalcified and the decalcification completely destroyed the RNA making it impossible to perform ISH. About 6 months ago, we switched to fixing our bone marrows in 10% NBF saturated with EDTA (this fixes and decals at the same time) with beautiful results with our IHC and also ISH. Even though our pathologists are elated that the IHC and ISH are great, they are not completely happy with the morphology of the H&E slides. I was just wondering if anyone has any suggestions so we can accomplish both great H&E morphology and great stains with IHC and ISH. Thanks, Charlene Henry, HT QIHC Histology/IHC Section Head Department of Pathology St. Jude Children's Research Hospital 332 North Lauderdale Memphis, TN 38105 Phone: 901-495-3349 FAX: 901-495-3100 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From georgecole <@t> ev1.net Sat May 15 10:16:53 2004 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] ALL OF THOSE VIRUS WARNINGS Message-ID: <000101c43a8f$a85c1d80$064dbad0@hppav> H EY!!! VIRUS PERSON WHO EVER YOU ARE----HHEEEYY YOU!!!---VIRUS ANDY!!!---COUNT YOUR DAYS---SOONER OR LATER YOU ARE GOING TO COME TO THIS HOSPITAL OR ANOTHER LIKE IT IN DESPAIR OF YOUR LIFE, AND IF THERE IS JUSTICE IN THE UNIVERSE---YOUR TREATMENT WILL BE CURTAILED-HELD UP---GARBLED!!--- BY A VIRUS-BLASTED COMPUTER SYSTEM. MAY YOU LIVE TO SEE THE DAY!!! georgecole@ev1.net From RSRICHMOND <@t> aol.com Sat May 15 13:24:04 2004 From: RSRICHMOND <@t> aol.com (RSRICHMOND@aol.com) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Retic nomenclature Message-ID: <134.2f167da4.2dd7ba44@aol.com> Lance Erickson at Primary Children's Medical Center in Salt Lake City Utah asks about the appropriate name for the reticulum stain, Gordon and Sweets or any other. The historically sanctioned name is reticulum stain, and there is no reason to change it. Thirty or forty years ago "reticulin" was sometimes considered to be a specific tissue component that the stain demonstrated, but since this is no longer thought to be the case the word "reticulin" should be abandoned. - Supposedly the PAS stain has similar specificity. Reticulum stains go back to the Chandler Foot era of surgical pathology (1930's-early 1940's), when it was thought that special stains would revolutionize surgical pathology in the way that immunohistochemistry actually did in the 1980's. By the late 1960's the stain was out of use in many large centers; I don't think I ever saw one when I was at Johns Hopkins between 1964 and 1972. I always thought of it as an old codger's stain when I was young. Now that I AM an old codger, I'm surprised it's still around. Bob Richmond Samurai Pathologist Gastonia NC From jstaruk <@t> masshistology.com Sat May 15 15:47:32 2004 From: jstaruk <@t> masshistology.com (Mass Histology Service) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Tissue processor repair in New England In-Reply-To: <134.2f167da4.2dd7ba44@aol.com> Message-ID: I searched the archives for the answer to this question but nothing showed up. Anyone know who to call in New England to repair a VIP? Thanks in advance. (Looking for free advice only. Those who plan to invoice me for answering this question need not reply.) Jim ____________________________ James E. Staruk, HT(ASCP) Mass Histology Service www.masshistology.com From katri <@t> cogeco.ca Sat May 15 17:43:07 2004 From: katri <@t> cogeco.ca ( Katri Tuomala) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Fixation and Decalcification of Bone Marrow Bxs. References: <5CB39BCA5724F349BCB748675C6CA1A231582D@SJMEMXMB02.stjude.sjcrh.local> Message-ID: <013601c43acd$fe88c760$ce989618@hala4.on.cogeco.ca> Hi Charlene, I think, if you fixed first with the 10% NBF properly and THEN decalcified with EDTA, your morphology would be great and EDTA decalcification does not affect staining or imuunohistochemistry. The reason people do these things together is to cut down the time, but you always pay for it. In this case with poor morphology. Cheers! Katri Katti Tuomala Dept. of Pathology St.Josep's Health Care Hamilton, Ontario, Canada ----- Original Message ----- From: "Henry, Charlene" To: Sent: Friday, May 14, 2004 4:34 PM Subject: [Histonet] Fixation and Decalcification of Bone Marrow Bxs. Fellow Histotechs, I would like to get some feedback on fixatives and decalcification protocols used in your laboratories. Our pathologists, like many, want perfection; so here is my problem. We were fixing our bone marrow bxs. in 10% NBF and then decalcification for 1 hour prior to processing. Because of the facility I work at, we perform IHC and ISH on a large percentage of our bxs. With the method we were using, some of our antibodies would not work on tissue that had been decalcified and the decalcification completely destroyed the RNA making it impossible to perform ISH. About 6 months ago, we switched to fixing our bone marrows in 10% NBF saturated with EDTA (this fixes and decals at the same time) with beautiful results with our IHC and also ISH. Even though our pathologists are elated that the IHC and ISH are great, they are not completely happy with the morphology of the H&E slides. I was just wondering if anyone has any suggestions so we can accomplish both great H&E morphology and great stains with IHC and ISH. Thanks, Charlene Henry, HT QIHC Histology/IHC Section Head Department of Pathology St. Jude Children's Research Hospital 332 North Lauderdale Memphis, TN 38105 Phone: 901-495-3349 FAX: 901-495-3100 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From madaryhtl <@t> juno.com Sat May 15 20:24:54 2004 From: madaryhtl <@t> juno.com (madaryhtl@juno.com) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] (no subject) Message-ID: <20040515.182537.29215.256284@webmail12.lax.untd.com> I use Kimwipes or cut up telephone book pages to clean of my wawa bath. Does anyone know what would be the best place to go to find out the value of older equipment for resale, or purchase? Specifically I have some average microtome and a decent embedder and an older buyt working processor that I cannot seem to find any onfo on regardiong value. The copmany is in the midst of downsizing, we lost a couple of histotechs, and are now getting rid of the equipment. I didn;t realize histotechs got laid off! How naive am I? From marktarango <@t> earthlink.net Sat May 15 21:13:08 2004 From: marktarango <@t> earthlink.net (Mark Adam Tarango) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Re: How do you clean your waterbaths? In-Reply-To: <200405151001.1bp2xF6Ml3NZFji0@eagle> Message-ID: <000001c43aeb$57e1d500$2702a8c0@markie> Hi Cathy, If you're looking to save some money, I've heard that some techs use cut up phone book pages. I've never done it, but I hear it works great. I've also seen more than one person use that plastic, cellophane wrap that you find when you open a new box of slides. They don't use it just once, it is kept on the edge of their waterbath and used many times over before being discarded and starting with a new piece. I've done this once or twice, it seem like the paraffin loves sticking to the stuff...like static cling or something..... I hope you find something that works well. Mark Tarango, HT(ASCP) AA Histologist Long Beach Memorial Medical Center PH-562-933-0761 marktarango@earthlink.net >>> Stevens Cathy 05/13/04 02:49PM >>> Just looking to see what various people use to clean the debris off their water baths while cutting. We use kimwipes. Anything better? Thanks Cathy Stevens H.T.(ASCP) BS Pathology Coordinator The Medical Center of Aurora P-303-695-2636 F-303-873-5660 cathy.stevens@healthonecares.com From t-sherman <@t> comcast.net Sat May 15 21:18:53 2004 From: t-sherman <@t> comcast.net (Todd Sherman) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Re: Fixative and H2O2 for frozen Message-ID: <40A6CF8D.2060100@comcast.net> -----BEGIN PGP SIGNED MESSAGE----- Hash: SHA1 Hello Jorge, Any H2O2 solution should be prepped daily - actually immediately before use. The peroxide oxidizes so quickly that it will lose its efficacy in short order; after all, you need lots of readily available substrate for the peroxidases to get optimally "quenched". Also, as far as solutions go, it's cheap and easy to make. Store the PBS. Don't store PBS/H2O2 solutions. The acetone:ethanol is probably more stable and could likely stand for a while; my question, why take the chance? It, too, is very easy, because they (X-CH2-COH-CH3 and CH3-CH2OH are polar molecules) are so soluble, and inexpensive to make. Further, fresh solution allows consistency because one eliminates another factor of variability from the day's "experimentation". If you are doing research, that's all the reason you need to make things fresh whenever possible. Other experts may disagree. Hope this helps, Todd Todd Sherman President HistoSoft Corporation "Biology in a new form..." Home: www.histosoft.com Member Services: www.myhistosoft.com histonet-request@lists.utsouthwestern.edu wrote: | | Today's Topics: | | 25. Fixative and H2O2 for frozen (Jorge Ivan Zapata Valencia) | | | ---------------------------------------------------------------------- | | Message: 25 | Date: Sat, 15 May 2004 07:46:00 +0000 | From: "Jorge Ivan Zapata Valencia" | Subject: [Histonet] Fixative and H2O2 for frozen | To: histonet@lists.utsouthwestern.edu | Message-ID: | Content-Type: text/plain; format=flowed | | Hello everybody | a technical question: may I use the same acetone:etanol mix to fix samples | different days, or do I have to prepare it feshly every day?? The same with | the H2O2 PBS solution for endogenous peroxidase blocking. Is it mandatory to | use fresh one? | Thank You very much | | jorge ivan zapata | | ------------------------------ -----BEGIN PGP SIGNATURE----- Version: GnuPG v1.2.3 (MingW32) Comment: Using GnuPG with MultiZilla - http://enigmail.mozdev.org iD8DBQFAps+LEmHXdrslGRcRAsRLAJ9aox6Pn9TRnZdvx/EaZNvqWz3TYwCg0N45 wQphw09emMfZt4mNVGCK8hM= =gQ1J -----END PGP SIGNATURE----- From carl.hobbs <@t> kcl.ac.uk Sun May 16 11:31:19 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] re rules?regs Message-ID: <000c01c43b63$37f12c50$95412cd9@home> Just read my post...comes across as quite rude....my apologies. Not intended that way at all. Different subject altogether. I agree that many quidelines/recommendadions are there for just that , so may require interpretation in their execution. Some people don't see it that way, tho and attempt to stick to "the letter". --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.684 / Virus Database: 446 - Release Date: 13/05/2004 From peptolab <@t> hamptons.com Sun May 16 12:25:39 2004 From: peptolab <@t> hamptons.com (peptolab) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Getting rid of used stuff Message-ID: <001f01c43b6a$cf9028c0$687bbd18@JEFF> Mary- try Belair Instrument Co., Inc. dist compound & stereo microscopes, tool makers Belair Instrument Co., Inc. 36 Commerce St. Springfield, NJ 7081 USA Phone: 973-912-8900 Fax: 973-374-2779 they always buy my used stuff and pick it up too. Jeff Silverman From Lizbeth_Kelly <@t> hgsi.com Sun May 16 15:17:51 2004 From: Lizbeth_Kelly <@t> hgsi.com (Lizbeth_Kelly@hgsi.com) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] NSH Region II Symposium (June 3,4, & 5th) Message-ID: ----- Forwarded by Lizbeth Kelly/Hgsi on 05/14/04 04:46 PM ----- Lizbeth Kelly 03/17/04 03:08 PM To: "'histonetters'" cc: TerDec@northarundel.org Subject: NSH Region II Symposium (June 3,4, & 5th) The Maryland Society of Histotechnologists (MSH) is sponsoring the Region II Symposium June 3, 4, and 1/2 day on the 5th at the Holiday Inn Select, North-Baltimore, Maryland. For information, you may contact Terri DeCarli at 410-787-4546 or Renate Jaacks at 410-879-9012. Lizbeth Kelly, HT (ASCP), QIHC Board Member, MSH From ThisisAnn <@t> aol.com Sun May 16 18:03:37 2004 From: ThisisAnn <@t> aol.com (ThisisAnn@aol.com) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] F/T Position in NJ Message-ID: <55.57a60a59.2dd94d49@aol.com> Labcorp, Inc., located in Raritan, NJ, is looking for an HT (ASCP), minimum of 2 years experience. Contact Ann Angelo, 908-526-2400 x2912 and/or fax your resume to 908-927-1129, attn: Ann Angelo From Megan.Kear <@t> hunter.health.nsw.gov.au Sun May 16 18:08:12 2004 From: Megan.Kear <@t> hunter.health.nsw.gov.au (Megan Kear) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] H&E stain problems Message-ID: We are currently having major problems with our H&E stain. We use a regressive hx and a non-alc eosin. I'd be greatful for any hints. We stain about 600+ slides /day and i'm finding we need to change the acid/alc, scot's blue, eosin & alcohols 3time/day. The automatic stainer is a Medite (Hacker). If anyone has a great methods for processing large quantities of slides or names of hx and eosin that are easy to work with i'd be greatful. Our lab is in Australia, NSW. Thanks, Ros. From kennedya <@t> email.cs.nsw.gov.au Sun May 16 19:29:17 2004 From: kennedya <@t> email.cs.nsw.gov.au (Andrew Kennedy) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Aussies and Kiwis standing up at microtomes Message-ID: <200405171027886.SM01316@crgcsls813> Hi Tim and Fred (and the rest of Histonetters too!!) In my experience, we Aussies don't like to stand up at the microtome if we can help it. Standing and cutting sections makes no sense at all - we might spill our beer and not be able to see the television that sits on the bench next to us so we can watch Aussie Rules (I prefer Rugby League, by the way) :-) But seriously folks, you would have to have high benches and even then you would probably hunch over the microtome - I remember seeing a histotech cutting standing up years ago when I was a lab assistant. She turned the microtome sideways, stood at the bench (which was desk height) and stooped over the microtome. That can't be good for your back! It was the most uncomfortable looking cutting position I've ever seen. Overlapping with the safety thread that is currently quite "hot", we really owe it to ourselves and to those we work with to be safe all the time! In Australia, Occupational Health and Safety is an important issue that affects everything we do in the workplace. However, any safety procedure that has been implemented in our labs is a result of consultation with the parties at stake. Safe Operating Procedures are designed by the people who will use them, not by some beaurocrat sitting in an office telling others what to do! Take control of your safety and use common sense at the same time! Makes sense to me!! Andrew Kennedy Senior Science Officer Anatomical Pathology Concord Repatriation General Hospital Hospital Road Concord NSW Australia 2139 ph: +612 9767 6115 Fax +612 9767 8427 "oderint dum metuant" From lpwenk <@t> covad.net Sun May 16 19:32:44 2004 From: lpwenk <@t> covad.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] HT Practical Grading References: <5C6CBCCEB04B894C8BD15B312487F7B201A330FE@ftwex01.txhealth.org> Message-ID: <008f01c43ba6$7fc2cfa0$41a7ff44@domainnotset.invalid> About 3-4 months after the deadline for turning in the practicals. This is what I tell my students. The practical deadline for this cycle was the end of March. So your students should expect to get the results of the practical anywhere from the end of June to the end of July. Much of it depends on how many sets were turned in from across the US, and how long it takes ASCP to validate the grading of the number of sets. The more sets, the longer it takes. (Grading normally occurs 6-8 weeks after the deadline, and then it is another 6-8 weeks for the validation of the grading and the letters to go out.) Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Willis, Donna" To: "'HistoNet'" Sent: Friday, May 14, 2004 2:50 PM Subject: [Histonet] HT Practical Grading > Anyone know when the pass/fail notices will be sent to those that have > recently taken their practical. I have 4 applicants that are eagerly > waiting for notification. > > Donna Willis > Histology Lab Manager > Harris Methodist Fort Worth, Tx > > > > The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From JNocito <@t> Pathreflab.com Mon May 17 05:34:52 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] VIP Tissue Processor malfunctioning..... In-Reply-To: <000001c43a07$b6b0d200$2702a8c0@markie> Message-ID: Mark, If you have a VIP 5, I just went through that. A hose in the back was plugged by paraffin. According to the guy who repaired it, this seems to be normal for the VIP 5. Joe Nocito -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Mark Adam Tarango Sent: Friday, May 14, 2004 6:04 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] VIP Tissue Processor malfunctioning..... Has anyone ever had a problem w/ their VIP processor pumping water into the last paraffin bath or retort chamber? This has happened four or five times in the last three months. It seems to always happen after I change the reagents. Am I to blame? P.S. What is the water for? Is its purpose to minimize fumes? I've heard someone say it's used to pump reagents in and out....something related to the vacuum and pressure, but this doesn't make sense to me... Thanks Mark Tarango _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cgfields <@t> lexhealth.org Mon May 17 06:33:00 2004 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Tissue processor repair in New England Message-ID: Southeast Pathology Instrument Service (1-843-588-2559) goes all the way to Puerto Rico ...maybe he goes to New England. He is in the Charleston area. His name is Michael Dietrich and he is great. Trained on Tissue tek products, Microm, microtomes, cryostats, etc. Give him a call and check it out. Carole Fields Lex Med Ctn W.Columbia, SC > -----Original Message----- > From: Mass Histology Service [SMTP:jstaruk@masshistology.com] > Sent: Saturday, May 15, 2004 4:48 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Tissue processor repair in New England > > I searched the archives for the answer to this question but nothing showed > up. Anyone know who to call in New England to repair a VIP? > > Thanks in advance. > > (Looking for free advice only. Those who plan to invoice me for answering > this question need not reply.) > > Jim > > ____________________________ > James E. Staruk, HT(ASCP) > Mass Histology Service > www.masshistology.com > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From mbecker <@t> pathlabinc.com Mon May 17 06:56:00 2004 From: mbecker <@t> pathlabinc.com (Michelle Becker) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Tissue processor repair in New England In-Reply-To: Message-ID: Have you tried New England Biomedical Services? Brian Hurley has serviced our VIP, microtomes, stainer, coverslipper etc. for many years. He is local and does a very thorough job. (781) 331-8642 NEBS@comcast.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Mass Histology Service Sent: Saturday, May 15, 2004 3:48 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tissue processor repair in New England I searched the archives for the answer to this question but nothing showed up. Anyone know who to call in New England to repair a VIP? Thanks in advance. (Looking for free advice only. Those who plan to invoice me for answering this question need not reply.) Jim ____________________________ James E. Staruk, HT(ASCP) Mass Histology Service www.masshistology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JMacDonald <@t> mtsac.edu Mon May 17 02:05:11 2004 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] VIP Tissue Processor malfunctioning..... Message-ID: The rotary valve goes out on some tissue processors. This allows the formalin into the paraffin bath. Jennifer From taylor.robin.e <@t> gene.com Mon May 17 07:52:10 2004 From: taylor.robin.e <@t> gene.com (Robin E. Taylor (Pathology)) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] EM Contract Labs? Message-ID: <40A8B57A.F912304@gene.com> Does anyone know of any contract labs on the west coast that do EM (TEM & SEM)? Thanks. Robin From ESerrano <@t> imim.es Mon May 17 08:24:37 2004 From: ESerrano <@t> imim.es (Serrano del Pozo, Erika) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Transportation of frozen sections Message-ID: <10708933FD0CD511BCFE000102BE5F72025FAD21@hpserv.imim.es> Hi Jorge Ivan, If you want to perform some immunohistochemistry or tunnel assay, sections must keep frozen, so you should make a shippment to Colombia with dry ice. If they defrost, proteins will degradate and it woudn?t be able to perform a good antigen detection (IHC). No idea about ordinary stainings... depending what you want to detect, I think better not defrost. Erika Serrano Research Assistant Centre de Regulaci? Gen?mica (CRG) Barcelona -SPAIN- > ---------- > From: Jorge Ivan Zapata Valencia > Sent: Saturday, May 15, 2004 3:33 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Transportation of frozen sections > > Hello everybody > I want to ask how can I transport frozen sections between two distant > places? I am now in Japan but I have to go back to Colombia soon and > finish > there the staining protocols I have to do. How I can transport the slides > in > such a way that my samples arrive to Colombia in good condition to be > stained? Must i fix them and keep them at room temperature? > Any suggestion is really appreciated > Thank You very much > Regards, > > JORGE IVAN ZAPATA > > _________________________________________________________________ > MSN 8 helps eliminate e-mail viruses. Get 2 months FREE*. > http://join.msn.com/?page=features/virus > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From pstefanova <@t> sten.sunnybrook.utoronto.ca Mon May 17 08:44:40 2004 From: pstefanova <@t> sten.sunnybrook.utoronto.ca (Petia P Stefanova) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] H&E stain problems References: Message-ID: <007001c43c15$1f750900$9d194c8e@WS21203> Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia From Carolyn.Earley <@t> leica-microsystems.com Mon May 17 09:24:44 2004 From: Carolyn.Earley <@t> leica-microsystems.com (Carolyn.Earley@leica-microsystems.com) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] (no subject) Message-ID: Hi Venita, I wanted to clarify to you that the current enclosed Leica Tissue Processor is the ASP300. The TP1050 is our last generation processor. I will be happy to forward information to you upon your request. Good luck in your search! Regards, Carolyn Earley Marketing Manager Leica Microsystems 847-405-7014 Carolyn.Earley@Leica-Microsystems.com Venita Capaldo To: histonet@lists.utsouthwestern.edu Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] (no subject) western.edu 05/13/2004 04:50 PM Hi there all. My lab is looking into purchasing a new processor and I would love some input. We are considering the Shandon Excelsior, the Tissue-Tek VIP5 and the Leica TP1050. If anyone has any input on these processors It would be greatly appreciated. We are a small dermatopathology lab and we routinly process 200-300 specimens per week. All of the specimens are skin or soft tissue and most are only 1mm to 5mm in size, but we do do some larger specimens as well. Thanks in advance for your thoughts! Sincerely, Venita Capaldo Christinmyhart@yahoo.com Allergy and Dermatology Specialists, Inc. Phoenix, Arizona (602)277-7686 --------------------------------- Do you Yahoo!? Yahoo! Movies - Buy advance tickets for 'Shrek 2' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JSCHUMA1 <@t> FAIRVIEW.ORG Mon May 17 12:23:36 2004 From: JSCHUMA1 <@t> FAIRVIEW.ORG (JENNIFER SCHUMACHER) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] Embedding centers--thanks Message-ID: Thank you to everyone who replied to my request about embedding centers. Jennifer 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 gudrun.lang <@t> aon.at Mon May 17 12:51:35 2004 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] eosin Message-ID: <003501c43c37$9900c990$eeeea8c0@SERVER> Hi netters (by the way "nett" in German means "friendly and nice", that fits) Can anyone tell me what pH a 2% Eosinsolution in Aqua dest. without any addition should have? Last time we measured ours with pH-meter it was around pH=8, what amazed me. An anionic dye in water is basic? If you prepare an eosinsolution, do you immidiatly add the acetic acid or just before use? thanks Gudrun Lang From ghoye <@t> iupui.edu Mon May 17 12:36:54 2004 From: ghoye <@t> iupui.edu (Hoye, Glenda F. (Fka Hood)) Date: Fri Sep 16 15:22:58 2005 Subject: [Histonet] contact Message-ID: <8FB28C0B73BA7E44B183083581C16092C94DE1@iu-mssg-mbx02.exchange.iu.edu> Hello, Could you please forward the contact information for Teresa Schuldt to me? She's in Norman, OK. I need a phone number or email if you have them available. gh Glenda Hoye, B.S., HT(ASCP) Histotechnology Program Director Indiana University School of Medicine Dept of Pathology & Laboratory Medicine 317-278-1599 From TJJ <@t> Stowers-Institute.org Mon May 17 13:11:13 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] VIP 5 malfunctions Message-ID: Nancy Klemme from Sakura? Can you address this please? I too have had a problem with water contaminating my solutions. First time it happened we were told the water filter bottle was too full and it siphoned up into the back of the instrument. We've made sure since then the water filter bottle was only about 1/4 full, and have had it empty out on another occasion. One of our bottles of alcohol was full, and we had water into the spill tray. There seems to be no reason for this that I can find. We are keeping a very close watch on our solutions to ensure we don't have contamination because I don't fully trust it not to malfunction. Joe Nocito says they had a paraffin plug in the back which caused their problems, and Mark Tarango had issues after he changed solutions. I can imagine that Joe's problem resulted from a bad rotary valve. Is this all operator error, or is something else going on here with the water filter? The rotary valve shouldn't have any involvement with the water filter, correct? To answer your question, Mark, the water filter just is an additional method of filtering the alcohol and xylene fumes and activates every time there is a pump in or out of these solutions. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From Rcartun <@t> harthosp.org Mon May 17 13:22:21 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Slides & blocks - Biohazard? Message-ID: Are glass microscopic slides and paraffin tissue blocks considered "biohazards"? We occasionally receive either unstained slides or a paraffin tissue block in a "Biohazard" bag from another institution for special testing. Technically speaking, the only personnel allowed to handle those materials would be those individuals wearing gloves. Richard Cartun From Jenny-Oblander <@t> omrf.ouhsc.edu Mon May 17 15:13:34 2004 From: Jenny-Oblander <@t> omrf.ouhsc.edu (Jenny Oblander) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] contact Message-ID: You can email her at tschuldt@rose.edu. Jenny -----Original Message----- From: Hoye, Glenda F. (Fka Hood) [mailto:ghoye@iupui.edu] Sent: Monday, May 17, 2004 12:37 PM To: Histonet@pathology.swmed.edu Subject: [Histonet] contact Hello, Could you please forward the contact information for Teresa Schuldt to me? She's in Norman, OK. I need a phone number or email if you have them available. gh Glenda Hoye, B.S., HT(ASCP) Histotechnology Program Director Indiana University School of Medicine Dept of Pathology & Laboratory Medicine 317-278-1599 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kwittle <@t> jhmi.edu Mon May 17 15:29:52 2004 From: kwittle <@t> jhmi.edu (Karen Wittler) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] VIP Processors Message-ID: Paraffin plug in the tubing? More often than not that was caused by over filling the paraffin containers. The vacuum pump will pump for three minutes and does not sense when the retort is full or empty completely. The tubing fills with paraffin and hardens. I believe the heating element is between retort chamber and the rotary valve, so the tubing is at room temperature. We have found that a little less solution is better than more. Stay beneath the lines, the lines are our friends. Water is part of the fume control system. Do we like our VIP's? We have two, year old VIP5s, one VIP 2000 and five VIP 3000s. We just had to learn how to care for them. Karen Wittler, HT(ASCP) Johns Hopkins Hospital Baltimore, MD From nena.dimaano <@t> stryker.com Mon May 17 15:47:43 2004 From: nena.dimaano <@t> stryker.com (Dimaano, Nena) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Acetabular components Message-ID: <20BE9059B6FC2D4B9E8FF2A9E2C4EEBD13F29D@HOS2KEXCHCL.howost.strykercorp.com> Hi everyone, I am looking to find protocols or references for sectioning and trimming acetabular components. thanks, Nena Dimaano Advanced Technology Stryker Orthopaedics 325 Corporate Drive Mahwah, NJ 07430 tel: 201-831-5338 fax: 201-831-6224 email: Nena.Dimaano@stryker.com From John.Garlits <@t> STJUDE.ORG Mon May 17 16:04:28 2004 From: John.Garlits <@t> STJUDE.ORG (Garlits, John) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] recommended biotinylated secondary for mouse monoclonal on human tissue? Message-ID: <1E0CC447E59C974CA5C7160D2A2854EC238AF7@SJMEMXMB04.stjude.sjcrh.local> Hello Histonetters, I was wondering if anyone could recommend a good detection system for a monoclonal mouse antibody to be used on human tissues (a new combination for me). This is for a PTH receptor antibody, and I have breast cancer slides as positive controls. I am thinking to use a biotinylated goat anti-mouse, since I have had good success using a biotinylated goat anti-rabbit for our GFP staining. Thank you! From jizv66 <@t> hotmail.com Mon May 17 18:35:38 2004 From: jizv66 <@t> hotmail.com (Jorge Ivan Zapata Valencia) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Transportation of frozen sections Message-ID: Apreciada Erika Mil gracias por su amable respuesta. Tengo que averiguar con la aerolinea si puedo usar Hielo seco, pues algunas no permiten transportar materiales con hielo seco. Mis mejores deseos, JORGE IVAN ZAPATA _________________________________________________________________ Add photos to your e-mail with MSN 8. Get 2 months FREE*. http://join.msn.com/?page=features/featuredemail From jizv66 <@t> hotmail.com Mon May 17 19:05:55 2004 From: jizv66 <@t> hotmail.com (Jorge Ivan Zapata Valencia) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Eosinophils with H&E Message-ID: Hello everybody I need to count eosinophils in eosinophilic granulomas from mouse infected with Toxocara canis. Does any body has a nice pic of mouse eosinophils stained with H&E? How can I distiguish them from Neutrophils? Thank You very much. JORGE IVAN ZAPATA _________________________________________________________________ Help STOP SPAM with the new MSN 8 and get 2 months FREE* http://join.msn.com/?page=features/junkmail From caroline.stott <@t> anatomy.otago.ac.nz Mon May 17 21:05:24 2004 From: caroline.stott <@t> anatomy.otago.ac.nz (Caroline Stott) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Aussies and Kiwis standing up at microtomes Message-ID: <5.2.1.1.0.20040518140054.00a8ed90@anatomy.otago.ac.nz> Hi Im in New Zealand and we sit down at the microtome. So Im not sure what this is about???? My boss only stands when his sciatica is playing up. But I have to say microtomes are an ergonomic nightmare. Caroline Caroline Stott Histology Service Unit Medical School University of Otago Dunedin (03) 479 7152 From lpwenk <@t> covad.net Mon May 17 20:46:16 2004 From: lpwenk <@t> covad.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Histology bench ergonomics References: Message-ID: <005001c43c79$ea0015c0$1337d445@domainnotset.invalid> An ergonomics seminar I went to said the workstation should be at elbow height. (See #2 below for caveat.) If you are going to be standing, then that is in the range of 33-45 inches If you are going to be sitting, then the height should be in the range of 26-34 inches. Why the range? 1. Line up a room of people, and have them stand with their upper arms hanging down at their sides, and the lower arms making an "L" in front of them, palms up. The elbow is at a 90 degree angle, and the forearms are parallel to the floor. Look at the height from the floor of everyone's arms.Wide range of distance from the floor. The same would apply to someone sitting down. 2. The "workstation" is not just the top of the counter. The height of the "thing" you are working on also has to be included in the height of the workstation. In other words, right now, I'm typing on a key board, and the extra 1 inch depth of the keyboard has to be added onto the height of the countertop. The height that people grab the coplin jar must be included in the overall height of the staining counter. The height that the microtome wheel handle is grabbed must be included in the overall height of the microtomy table. If these additional heights are not included, then if a 30 inch workbench is "perfect" for someone to sit and have their arms rest on the top of the counter at the 90 degree/parallel to the floor position, then when the microtome handle is added, the person has to "reach up", thereby being forced to raise their shoulders. This is not good ergonomically. So, how does a lab construct a working laboratory with people ranging in height from 4'10" to 6'2"? Well, either construct for 95% of the people (get a happy median), or buy the counters that can be adjusted up and down in height. We have those for our grossing stations, because different residents and pathologists are using the same stations, and we needed that ability to adjust the heights. And the PA's, who are there 8 hours a day, and adjust their grossing stations just for themselves, and can alter it when they decide to sit instead of stand. Hope this helps. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Thursday, May 13, 2004 11:11 AM Subject: [Histonet] Histology bench ergonomics > Does anyone have any data or information on bench height for microtomy? I > know a lot of people cut at routine lab bench height (36" I think) with no > trouble, and some cut at desk high benches (30"). Is there any > safety/ergonomic guidelines or opinions as to which is better for > microtomy? I actually prefer desk height, so I can keep my feet on the > floor - I'm just wondering if there is anything official out there - I've > looked at a couple of sites, but can't find a lot to support my idea that > desk height is better. > > Thanks - > Jackie O' > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From alexander.nader <@t> wgkk.sozvers.at Mon May 17 01:52:32 2004 From: alexander.nader <@t> wgkk.sozvers.at (Nader, Alexander) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Fixation and Decalcification of Bone Marrow Bxs. Message-ID: <4FE843CBBAC1D211A8150008C70952DAAE7E01@hk01nt05.hkh.wgkk.sozvers.at> > About 6 months ago, we switched to fixing our bone marrows > in 10% NBF saturated with EDTA (this fixes and decals at the > same time) > with beautiful results with our IHC and also ISH. Even though our > pathologists are elated that the IHC and ISH are great, they are not > completely happy with the morphology of the H&E slides. I was just > wondering if anyone has any suggestions so we can accomplish > both great H&E morphology and great stains with IHC and ISH. What is your formula for the NBF-EDTA mixture? pH? Alexander Nader MD Vienna, Austria From research <@t> nhls.ac.za Tue May 18 03:26:00 2004 From: research <@t> nhls.ac.za (Research) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Mismatch repair protein IHC Message-ID: Can anyone provide help / protocol with immunohistochemistry for MLH1, MSH2, MSH6 and PMS2? Research Laboratory Department of Anatomical Pathology South African Institute for Medical Research Johannesburg South Africa From Jamie.Dukes <@t> se.amedd.army.mil Tue May 18 05:53:00 2004 From: Jamie.Dukes <@t> se.amedd.army.mil (Dukes, Jamie Mr EAMC) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] pathology tech Message-ID: <4FB3076916FCD311AF7900805FA7A678077B4A8E@dasmthfdz001.amedd.army.mil> hello everyone, I have a problem with one of the pathologist at DDEAMC(Fort Gordon). She insist on having the path tech stand beside her to close the cassette for her when there is a ton of other work to do like keepin the morgue up to par. Can anyone help me explain to my department head that is right or safe because of too many hands handling small biopsies jamie dukes From celebrej <@t> HHSC.CA Tue May 18 06:21:34 2004 From: celebrej <@t> HHSC.CA (Celebre Julia) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Congo Red and Amyloid Message-ID: <3AADFB88753AD31189C100902786B91C0E41CA1D@hch_nt_exchange.hhsc.ca> Morning Histonetters..... Our lab is currently having a 'discussion' about our Congo Red stain for amyloid in regards to when Hematoxylin should be used, before or after. Is there anyone out there that can explain the reason it should be used prior or post the Congo Red step? Any information will be greatly appreciated. Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext46145 email: celebrej@hhsc.ca This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From MAUGER <@t> email.chop.edu Tue May 18 06:41:40 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] recommended biotinylated secondary for mousemonoclonal on human tissue? Message-ID: To John Garlits, Any biotinylated anti- mouse will do. I use horse anti ms from vector labs(biotinylated,with their ABC detection. Works beautifully. Jo >>> "Garlits, John" 05/17/04 05:04PM >>> Hello Histonetters, I was wondering if anyone could recommend a good detection system for a monoclonal mouse antibody to be used on human tissues (a new combination for me). This is for a PTH receptor antibody, and I have breast cancer slides as positive controls. I am thinking to use a biotinylated goat anti-mouse, since I have had good success using a biotinylated goat anti-rabbit for our GFP staining. Thank you! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jon.Roberts <@t> stjohn.org Tue May 18 07:57:52 2004 From: Jon.Roberts <@t> stjohn.org (Roberts, Jon) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] MIcrotome alignment Message-ID: <5B6EF454E075D94EB9DA2EB364476233010A52E2@hssjex11.SJHS.DS.sjhs.com> I have recently purchased a Leica RM2255 with the new precise specimen orientation system that adjust to an exact zero position or x-y axis standard. But I also have old leicas without this system. Is there any quick and easy way to align my old leicas to the new one???? Just wondering? I though I've heard of such a device, but wonder if anyone else has heard of it or actually used it? CONFIDENTIALITY NOTICE: This email message and any accompanying data are confidential, and intended only for the named recipient(s). If you are not the intended recipient(s), you are hereby notified that the dissemination, distribution, and or copying of this message is strictly prohibited. If you receive this message in error, or are not the named recipient(s), please notify the sender at the email address above, delete this email from your computer, and destroy any copies in any form immediately. From froyer <@t> bitstream.net Tue May 18 09:00:13 2004 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] MIcrotome alignment In-Reply-To: <5B6EF454E075D94EB9DA2EB364476233010A52E2@hssjex11.SJHS.DS.sjhs.com> References: <5B6EF454E075D94EB9DA2EB364476233010A52E2@hssjex11.SJHS.DS.sjhs.com> Message-ID: <40AA16ED.5050702@bitstream.net> Newcomer Supply, Madison, WI sells a microtome aligner. Product Code 6255A Web URL Check it out. ~ Ford Ford M. Royer, MT(ASCP) Analytical Instruments, llc Minneapolis, MN 55427 (800) 565-1895, Ext. 17 Roberts, Jon wrote: >I have recently purchased a Leica RM2255 with the new precise specimen orientation system that adjust to an exact zero position or x-y axis standard. But I also have old leicas without this system. Is there any quick and easy way to align my old leicas to the new one???? Just wondering? I though I've heard of such a device, but wonder if anyone else has heard of it or actually used it? > > >CONFIDENTIALITY NOTICE: This email message and any accompanying data are confidential, and intended only for the named recipient(s). If you are not the intended recipient(s), you are hereby notified that the dissemination, distribution, and or copying of this message is strictly prohibited. If you receive this message in error, or are not the named recipient(s), please notify the sender at the email address above, delete this email from your computer, and destroy any copies in any form immediately. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From jqb7 <@t> cdc.gov Tue May 18 08:58:27 2004 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:59 2005 Subject: FW: [Histonet] MIcrotome alignment Message-ID: From: Bartlett, Jeanine Sent: Tuesday, May 18, 2004 9:01 AM To: 'Roberts, Jon' Subject: RE: [Histonet] MIcrotome alignment Check out www.newcomersupply.com It's called a microtome aligner. Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Roberts, Jon Sent: Tuesday, May 18, 2004 8:58 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] MIcrotome alignment I have recently purchased a Leica RM2255 with the new precise specimen orientation system that adjust to an exact zero position or x-y axis standard. But I also have old leicas without this system. Is there any quick and easy way to align my old leicas to the new one???? Just wondering? I though I've heard of such a device, but wonder if anyone else has heard of it or actually used it? CONFIDENTIALITY NOTICE: This email message and any accompanying data are confidential, and intended only for the named recipient(s). If you are not the intended recipient(s), you are hereby notified that the dissemination, distribution, and or copying of this message is strictly prohibited. If you receive this message in error, or are not the named recipient(s), please notify the sender at the email address above, delete this email from your computer, and destroy any copies in any form immediately. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From convmcm <@t> cc.usu.edu Tue May 18 09:09:17 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] H&E stain problems In-Reply-To: <007001c43c15$1f750900$9d194c8e@WS21203> Message-ID: <000701c43ce1$b5f00250$4a737b81@Cygnus> We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From garygill <@t> dcla.com Tue May 18 09:34:49 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] H&E stain problems Message-ID: Other details being equal, hand staining will beat machine staining when slides are dipped in rinses repeatedly. Sakura DRS-2000 has dip ("agitation") feature. Other machines? Gary Gill -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Tuesday, May 18, 2004 9:09 AM To: 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ 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 Terry.Marshall <@t> rothgen.nhs.uk Tue May 18 09:34:49 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] H&E stain problems Message-ID: Connie remarks: "In truth, I prefer my hand stained sections better than when they're stained automatically." When I first saw what I call x-y stainers, I thought that we had in this, something that reproduced hand staining. Not so. What I think is lacking is the brisk agitation necessary to break down the interface between old and new solution. I'm not so sure about the rinsing either. 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 18 May 2004 15:09 To: 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ 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 akwilliams75 <@t> hotmail.com Tue May 18 09:31:19 2004 From: akwilliams75 <@t> hotmail.com (kevin williams) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Slides & blocks - Biohazard? Message-ID: Dear Richard: I have been told that shavings are not Biohazard, because they have been processed, so how can a single "shaving" on a slide be considered biohazard? Yours faithfully, A. Kevin Williams >From: "Richard Cartun" >To: >Subject: [Histonet] Slides & blocks - Biohazard? >Date: Mon, 17 May 2004 14:22:21 -0400 > >Are glass microscopic slides and paraffin tissue blocks considered >"biohazards"? We occasionally receive either unstained slides or a >paraffin tissue block in a "Biohazard" bag from another institution for >special testing. Technically speaking, the only personnel allowed to >handle those materials would be those individuals wearing gloves. > >Richard Cartun > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ [1]Stop worrying about overloading your inbox - get MSN Hotmail Extra Storage! References 1. http://g.msn.com/8HMBENUS/2737??PS=47575 From pstefanova <@t> sten.sunnybrook.utoronto.ca Tue May 18 09:59:27 2004 From: pstefanova <@t> sten.sunnybrook.utoronto.ca (Petia P Stefanova) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] H&E stain problems References: <000701c43ce1$b5f00250$4a737b81@Cygnus> Message-ID: <002701c43ce8$bbf7b970$9d194c8e@WS21203> Hi Connie, I use Leica autostainer XL and they offer even longer / 20 sec./ differentiation with acid alcohol in their manual. Have a good day! Petia ----- Original Message ----- From: "Connie McManus" To: "'Petia P Stefanova'" ; "'Megan Kear'" ; Sent: Tuesday, May 18, 2004 10:09 AM Subject: RE: [Histonet] H&E stain problems > We use almost the exact same protocol... we use Surgipath Harris, but we > prepare eosin in-house. One thing I am amazed at in this protocol is > the length of time in the acid alcohol. Do you use an autostainer? We > have a Leica. The time is set to 1 second in the acid ETOH and > sometimes the sections are almost too differentiated. I can't imagine > 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the > slides in and quickly put them in running water. I must have a very > strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my > hand stained sections better than when they're stained automatically. > > Just wondering and blabbering (hey, it's Tuesday, what do you expect??) > > Connie McManus > Utah Veterinary Diagnostics Laboratory > Utah State University > Logan, UT > Phone: 435/797-1891 > fax: 435/797-2805 > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P > Stefanova > Sent: Monday, May 17, 2004 6:45 AM > To: Megan Kear; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] H&E stain problems > > Hi, > > I use Harris's hematoxylin which is also regressive and purchase my > hematoxylin and eosin /alcohol-based/ from www.surgipath.com. > I get very good H&E staining with this protocol. > > REAGENT TIME > Xylene 3 min. > Xylene 3 min. > Abs. alc. 2 min. > Abs. alc. 2 min > 95% alc. 2 min > 80% alc. 2 min > Wash /tap water/ 30 sec. > Hematoxylin 8 min. > Wash /tap water/ 2 min. > Acid alcohol 6 sec. > Wash /tap water/ 2 min. > Wash /tap water/ 5 min > 80% alc. 30 sec. > Eosin 15 sec. > 95% alc. 10 sec. > Abs. alc. 30 sec. > Abs. alc. 30 sec. > Xylene 1 min. > Xylene 1 min. > Xylene Exit > > Hope it helps! > Petia > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Jackie.O'Connor <@t> abbott.com Tue May 18 09:49:55 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Slides & blocks - Biohazard? Message-ID: Prions. "kevin williams" Sent by: histonet-bounces@lists.utsouthwestern.edu 05/18/2004 09:31 AM To: Rcartun@harthosp.org, histonet@pathology.swmed.edu cc: Subject: RE: [Histonet] Slides & blocks - Biohazard? Dear Richard: I have been told that shavings are not Biohazard, because they have been processed, so how can a single "shaving" on a slide be considered biohazard? Yours faithfully, A. Kevin Williams >From: "Richard Cartun" >To: >Subject: [Histonet] Slides & blocks - Biohazard? >Date: Mon, 17 May 2004 14:22:21 -0400 > >Are glass microscopic slides and paraffin tissue blocks considered >"biohazards"? We occasionally receive either unstained slides or a >paraffin tissue block in a "Biohazard" bag from another institution for >special testing. Technically speaking, the only personnel allowed to >handle those materials would be those individuals wearing gloves. > >Richard Cartun > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ [1]Stop worrying about overloading your inbox - get MSN Hotmail Extra Storage! References 1. http://g.msn.com/8HMBENUS/2737??PS=47575 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From convmcm <@t> cc.usu.edu Tue May 18 10:41:18 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] H&E stain problems In-Reply-To: Message-ID: <001101c43cee$91151ee0$4a737b81@Cygnus> Wow. What a lot of interesting comments!! I agree with Terry re the agitation. When I watch the stainer do those dips (I can program how many, but NOT the briskness), I wonder if you could even call it agitation. My hand dips are very brisk. Also, I don't bother letting the slides stay in the alcohols for 1 minute or 2, I give the slides about 20 -30 good brisk dips in each solution, then the timed rinses & staining. This has always been far more satisfactory to me than those sllllooooowwwww dips from the machine. As for the kind of hematoxylin, someone suggested I throw out the Harris and do Gills III. I've tried Gill III before and I much prefer the Harris. So it's just a matter of personal preference on that... AND what your pathologist likes *G* Everyone having an nice Tuesday??? *g* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, May 18, 2004 7:35 AM To: Connie McManus; Petia P Stefanova; Megan Kear; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Connie remarks: "In truth, I prefer my hand stained sections better than when they're stained automatically." When I first saw what I call x-y stainers, I thought that we had in this, something that reproduced hand staining. Not so. What I think is lacking is the brisk agitation necessary to break down the interface between old and new solution. I'm not so sure about the rinsing either. 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 18 May 2004 15:09 To: 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ 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 garygill <@t> dcla.com Tue May 18 11:41:14 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] H&E stain problems Message-ID: Hello Terry: Terry's right. Agitation is essential to good staining. Dye that remains in cells and tissues represents the difference between what the dye solutions put in and the rinses take out. There is bound dye that remains, and loosely infiltrated dye among the protein molecules that will remain and reduce crispness and contrast unless removed by pronounced agitation. Some is better than none, and more is better than some. Some general rinse tips: * Keep rinse volume high/deep to minimize dilution * Rinse in series of at least 3 baths * Dip constantly, totally in-and-out about once per second * Keep rinses clean (e.g., change/rotate when last bath in a series begins to be colored) * For acid dyes such as eosin, water will extract dye most, methanol, ethanol, and isopropanol in decreasing degrees * 0.5% glacial acetic acid in water can be substituted effectively for 95% ethanol Rinsing is most effective when the difference in concentration between the dye in the cells and tissue and the dye in the rinses is greatest (i.e., difference in chemical potential). When the rinses are dirty (i.e., the concentration of dye in the rinse approaches/equals/exceeds that in the cells), there is no net movement of dye out of the substrate). To appreciate the value of rinsing, don't do it on an experimental basis. Take an extra smear or tissue section directly from eosin into absolute alcohol. I've done the same thing for the Pap stain, going directly from EA into absolute alcohol -- the staining results are unrecognizable. Gary Gill -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, May 18, 2004 9:35 AM To: Connie McManus; Petia P Stefanova; Megan Kear; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Connie remarks: "In truth, I prefer my hand stained sections better than when they're stained automatically." When I first saw what I call x-y stainers, I thought that we had in this, something that reproduced hand staining. Not so. What I think is lacking is the brisk agitation necessary to break down the interface between old and new solution. I'm not so sure about the rinsing either. 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 18 May 2004 15:09 To: 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ 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 mari.ann.mailhiot <@t> leica-microsystems.com Tue May 18 11:47:53 2004 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] H&E stain problems Message-ID: Connie Your acid alcohol may be too strong. Usually a .25% solution is strong enough. If you would like to discuss just give me a call. Regards Mari Ann Mailhiot BA HT ASCP Application Specialist Leica Technical Assistance Center 800 248 0123 x7267 847 236 3063 fax mari.ann.mailhiot@leica-microsystems.com www.leica-microsystems.com "Connie McManus" To: "'Petia P Stefanova'" , "'Megan Sent by: Kear'" , histonet-bounces@lists.utsouth cc: western.edu Subject: RE: [Histonet] H&E stain problems 05/18/2004 09:09 AM We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ 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 srishan <@t> mail.holyname.org Tue May 18 11:58:33 2004 From: srishan <@t> mail.holyname.org (srishan@mail.holyname.org) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] CJD DECONTAMINATION PROCEDURE Message-ID: Hello everyone, This question might have been posted before, but, Histology Department is expecting a brain biopsy from a possible CJD patient today. The specimen will be coming down in formalin from the O.R. It will be kept in formalin for 2-7 days and will be transferred to formic acid for the required time. Then the specimen will be transferred to formalin before processing for another 2 days. Has anyone dealt with this procedure before. These are the questions asked by the hstology department. How do you decontaminate the processor, microtome, stainer and coverslipper? What kind of protective gear apart from standard precaution has to be used? How about the shaving of the tissues? How do you dispose the formalin from the original formalin container where the specimen was sent in and how do you dispose the reagents from the processor and the stainer? Can this biopsy be processed, once treated with formic acid, along with the other specimens or should the biopsy be processed with the other specimens? Need Help!! Thank you in advance Mala Srishan Histology Supervisor Holy Name Hospital Teaneck, NJ 07666 From garygill <@t> dcla.com Tue May 18 12:03:43 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] H&E stain problems Message-ID: You may have heard that classical music was composed by dead white Europeans. Well, Harris hematoxylin was composed by a dead white American (physician at Jefferson Hospital in 1904). Gill hematoxylin was composed by a live white American. So if you want to liven things up, go with Gill's! Gary Gill (one and the same) PS -- No royalties involved, thanks to bad advice in 1972 from corporate counsel for Johns Hopkins Medical School. -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Tuesday, May 18, 2004 10:41 AM To: 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Wow. What a lot of interesting comments!! I agree with Terry re the agitation. When I watch the stainer do those dips (I can program how many, but NOT the briskness), I wonder if you could even call it agitation. My hand dips are very brisk. Also, I don't bother letting the slides stay in the alcohols for 1 minute or 2, I give the slides about 20 -30 good brisk dips in each solution, then the timed rinses & staining. This has always been far more satisfactory to me than those sllllooooowwwww dips from the machine. As for the kind of hematoxylin, someone suggested I throw out the Harris and do Gills III. I've tried Gill III before and I much prefer the Harris. So it's just a matter of personal preference on that... AND what your pathologist likes *G* Everyone having an nice Tuesday??? *g* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, May 18, 2004 7:35 AM To: Connie McManus; Petia P Stefanova; Megan Kear; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Connie remarks: "In truth, I prefer my hand stained sections better than when they're stained automatically." When I first saw what I call x-y stainers, I thought that we had in this, something that reproduced hand staining. Not so. What I think is lacking is the brisk agitation necessary to break down the interface between old and new solution. I'm not so sure about the rinsing either. 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 18 May 2004 15:09 To: 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ 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 Tue May 18 12:58:07 2004 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] CJD DECONTAMINATION PROCEDURE Message-ID: The WHO booklet was too large to send as an attachment so it couldn't be delivered. I will try and send you what I think is the most relevant chapter. Perhaps you could visit the WHO site and get more information if needed. In the meantime, the Surveillance Center link below should have enough information. Jeanine - Mala: There is a lot of conflicting information out there but here is information from 2 very credible sources for you to review. One is the National Surveillance Center at case Western and the other is the WHO. Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 http://www.cjdsurveillance.com and the WHO booklet is sent as an attachment. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of srishan@mail.holyname.org Sent: Tuesday, May 18, 2004 12:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CJD DECONTAMINATION PROCEDURE Hello everyone, This question might have been posted before, but, Histology Department is expecting a brain biopsy from a possible CJD patient today. The specimen will be coming down in formalin from the O.R. It will be kept in formalin for 2-7 days and will be transferred to formic acid for the required time. Then the specimen will be transferred to formalin before processing for another 2 days. Has anyone dealt with this procedure before. These are the questions asked by the hstology department. How do you decontaminate the processor, microtome, stainer and coverslipper? What kind of protective gear apart from standard precaution has to be used? How about the shaving of the tissues? How do you dispose the formalin from the original formalin container where the specimen was sent in and how do you dispose the reagents from the processor and the stainer? Can this biopsy be processed, once treated with formic acid, along with the other specimens or should the biopsy be processed with the other specimens? Need Help!! Thank you in advance Mala Srishan Histology Supervisor Holy Name Hospital Teaneck, NJ 07666 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kbradshaw <@t> lcpath.com Tue May 18 12:55:17 2004 From: kbradshaw <@t> lcpath.com (Kari Bradshaw) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] CJD DECONTAMINATION PROCEDURE In-Reply-To: Message-ID: These people have all the answers: National Prion Pathology Surveillance Center Case Western Reserve University 2085 Adelbert Road Cleveland,Ohio 44106 216-368-0587 www.cjdsurveillance.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of srishan@mail.holyname.org Sent: Tuesday, May 18, 2004 9:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CJD DECONTAMINATION PROCEDURE Hello everyone, This question might have been posted before, but, Histology Department is expecting a brain biopsy from a possible CJD patient today. The specimen will be coming down in formalin from the O.R. It will be kept in formalin for 2-7 days and will be transferred to formic acid for the required time. Then the specimen will be transferred to formalin before processing for another 2 days. Has anyone dealt with this procedure before. These are the questions asked by the hstology department. How do you decontaminate the processor, microtome, stainer and coverslipper? What kind of protective gear apart from standard precaution has to be used? How about the shaving of the tissues? How do you dispose the formalin from the original formalin container where the specimen was sent in and how do you dispose the reagents from the processor and the stainer? Can this biopsy be processed, once treated with formic acid, along with the other specimens or should the biopsy be processed with the other specimens? Need Help!! Thank you in advance Mala Srishan Histology Supervisor Holy Name Hospital Teaneck, NJ 07666 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dmnelson <@t> iastate.edu Tue May 18 13:03:49 2004 From: dmnelson <@t> iastate.edu (Diane M Nelson) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] research charges Message-ID: <6.1.0.6.2.20040518124918.01b5caa0@dmnelson.mail.iastate.edu> Hi Histonetters! Our veterinary lab is in the process of up dating our research charges. I would like to know what other labs/hospitals charge for H & E bone slides and H & E eye slides. Currently we are charging, Bone: $19.00 for decalcifying in 25% formic acid, process, embed, sectioning and H & E stain. We are losing money on our larger bone projects. Our charges for processing, embedding sectioning and H & E stain for eye slides is $9.00. It also depends on the project rather or not we make or lose money. The researchers do their own grossing of tissue. Any help would be greatly appreciated! Thanks in advance. Diane Gerjets Iowa State University From fmonson <@t> wcupa.edu Tue May 18 13:04:45 2004 From: fmonson <@t> wcupa.edu (Monson, Frederick ) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] CJD DECONTAMINATION PROCEDURE Message-ID: http://www.nursingceu.com/NCEU/courses/prions/ http://www.mad-cow.org/tonsil_test2.html#Biosafety A start. Cheers Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone/FAX: 610-738-0437 eMail: fmonson@wcupa.edu CASI Page and Scheduling http://darwin.wcupa.edu/CASI/ -----Original Message----- From: srishan@mail.holyname.org [mailto:srishan@mail.holyname.org] Sent: Tuesday, May 18, 2004 12:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CJD DECONTAMINATION PROCEDURE Hello everyone, This question might have been posted before, but, Histology Department is expecting a brain biopsy from a possible CJD patient today. The specimen will be coming down in formalin from the O.R. It will be kept in formalin for 2-7 days and will be transferred to formic acid for the required time. Then the specimen will be transferred to formalin before processing for another 2 days. Has anyone dealt with this procedure before. These are the questions asked by the hstology department. How do you decontaminate the processor, microtome, stainer and coverslipper? What kind of protective gear apart from standard precaution has to be used? How about the shaving of the tissues? How do you dispose the formalin from the original formalin container where the specimen was sent in and how do you dispose the reagents from the processor and the stainer? Can this biopsy be processed, once treated with formic acid, along with the other specimens or should the biopsy be processed with the other specimens? Need Help!! Thank you in advance Mala Srishan Histology Supervisor Holy Name Hospital Teaneck, NJ 07666 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Tue May 18 13:32:48 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Presto Fix - ? Message-ID: Anyone using "Presto Fix" for bone marrow core biopsies? If so, what does it do to immunoreactivity? Thank you. Richard Cartun From emry <@t> u.washington.edu Tue May 18 13:34:18 2004 From: emry <@t> u.washington.edu (P. Emry) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Aussies and Kiwis standing up at microtomes In-Reply-To: <200405171027886.SM01316@crgcsls813> References: <200405171027886.SM01316@crgcsls813> Message-ID: Ahhhhh working with a beer at my elbow. Sounds very sane to me. I think we got one of your country women..a Wealthal..up for a phd. She denies dwarf tossing, but I have heard things to the contrary. I have two square carts with drawers in used in the dental school. They have wheels on them. I put the microtome on one and the waterbath on the other. They sit side-by side with a space between. They are just high enough for me to use a standard height chair with wheels and move back and forth between them. It is the best arrangement I have found. I can get up over the water bath to get a good look at what is going on. I really like it. Not hard on the back like benches. Would suggest it for those having problems with benches and sitting still in one possition. If I can figure out how to put an ice bucket for the beer between, we's really have something. Trisha On Mon, 17 May 2004, Andrew Kennedy wrote: > Hi Tim and Fred (and the rest of Histonetters too!!) > > > > In my experience, we Aussies don't like to stand up at the microtome if we > can help it. Standing and cutting sections makes no sense at all - we might > spill our beer and not be able to see the television that sits on the bench > next to us so we can watch Aussie Rules (I prefer Rugby League, by the way) > > :-) > > But seriously folks, you would have to have high benches and even then you > would probably hunch over the microtome - I remember seeing a histotech > cutting standing up years ago when I was a lab assistant. She turned the > microtome sideways, stood at the bench (which was desk height) and stooped > over the microtome. That can't be good for your back! It was the most > uncomfortable looking cutting position I've ever seen. > > > > Overlapping with the safety thread that is currently quite "hot", we really > owe it to ourselves and to those we work with to be safe all the time! > > > > In Australia, Occupational Health and Safety is an important issue that > affects everything we do in the workplace. However, any safety procedure > that has been implemented in our labs is a result of consultation with the > parties at stake. Safe Operating Procedures are designed by the people who > will use them, not by some beaurocrat sitting in an office telling others > what to do! Take control of your safety and use common sense at the same > time! Makes sense to me!! > > > > Andrew Kennedy > > Senior Science Officer > > Anatomical Pathology > > Concord Repatriation General Hospital > > Hospital Road > Concord NSW Australia 2139 > > > > ph: +612 9767 6115 > > Fax +612 9767 8427 > > > > "oderint dum metuant" > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From kwittle <@t> jhmi.edu Tue May 18 15:29:47 2004 From: kwittle <@t> jhmi.edu (Karen Wittler) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Congo Red Message-ID: We stain our Congo Reds on an automated stainer now. Before that we stained with hematoxylin first because rinsing and differentiation was required and we used an alkaline Congo Red. After staining with the Congo Red in a saturated sodium chloride- alcohol solution,we did not rinse,but rapidly dehydrated, cleared and mounted For our H&Es we use Modified Harris Hematoxylin and Eosin-Y w/ Phloxine both manufactured by Richard Allen. Karen Wittler, HT(ASCP) Johns Hopkins Hospital Baltimore, MD From froyer <@t> bitstream.net Tue May 18 16:16:32 2004 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] MIcrotome alignment In-Reply-To: References: Message-ID: <40AA7D30.10601@bitstream.net> On their web site catalog, they show a price of $775.00. Contact Marcia Welch at: 800-383-7799 or Email: for a customer price quote. Ford M. Royer, MT(ASCP) Analytical Instruments, llc 1200 Mendelssohn Ave. N., Ste. 50 Minneapolis, MN 55427 (800) 565-1895, ext. 17 URL: KHays@mbhs.org wrote: > > Kathy Tedford-Hays HT (ASCP) > Technical Specialist, Histology Dept > (601)-968-3070 ext 7398 > Baptist Medical Center > 1225 North State Street > Jackson, MS 39202 > > how much is it , do you know? > > > > "Ford Royer" > Sent by: histonet-bounces@lists.utsouthwestern.edu > > 05/18/2004 09:00 AM > > > To: histonet@lists.utsouthwestern.edu > cc: > Fax to: > Subject: Re: [Histonet] MIcrotome alignment > > > > > Newcomer Supply, Madison, WI sells a microtome aligner. Product Code > 6255A > Web URL > Check it out. > > ~ Ford > Ford M. Royer, MT(ASCP) > Analytical Instruments, llc > Minneapolis, MN 55427 > (800) 565-1895, Ext. 17 > > > Roberts, Jon wrote: > > >I have recently purchased a Leica RM2255 with the new precise > specimen orientation system that adjust to an exact zero position or > x-y axis standard. But I also have old leicas without this system. > Is there any quick and easy way to align my old leicas to the new > one???? Just wondering? I though I've heard of such a device, but > wonder if anyone else has heard of it or actually used it? > > > > > >CONFIDENTIALITY NOTICE: This email message and any accompanying data > are confidential, and intended only for the named recipient(s). If > you are not the intended recipient(s), you are hereby notified that > the dissemination, distribution, and or copying of this message is > strictly prohibited. If you receive this message in error, or are not > the named recipient(s), please notify the sender at the email address > above, delete this email from your computer, and destroy any copies in > any form 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 > This internet email has been certified virus free by BHS Anti-Virus > system. > > From jstaruk <@t> masshistology.com Tue May 18 18:59:45 2004 From: jstaruk <@t> masshistology.com (Mass Histology Service) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] research charges In-Reply-To: <6.1.0.6.2.20040518124918.01b5caa0@dmnelson.mail.iastate.edu> Message-ID: Diane, We charge one price across the board, $5.50 if the tissue is received in a cassette and $2.50 if it needs decalcification. Our philosophy is that we don't charge less for simple specimens, so why charge more for occasional complicated ones? Of course if it's a large project consisting of many whole eyes or femur heads, I'd probably double our current prices to cover for the additional labor and material. Jim ____________________________ James E. Staruk, HT(ASCP) Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Diane M Nelson Sent: Tuesday, May 18, 2004 2:04 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] research charges Hi Histonetters! Our veterinary lab is in the process of up dating our research charges. I would like to know what other labs/hospitals charge for H & E bone slides and H & E eye slides. Currently we are charging, Bone: $19.00 for decalcifying in 25% formic acid, process, embed, sectioning and H & E stain. We are losing money on our larger bone projects. Our charges for processing, embedding sectioning and H & E stain for eye slides is $9.00. It also depends on the project rather or not we make or lose money. The researchers do their own grossing of tissue. Any help would be greatly appreciated! Thanks in advance. Diane Gerjets Iowa State University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Tue May 18 20:08:30 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Efficiency of p o and m xylenes in dewaxing Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E1C8@simba.kids> Hi all, Would anyone have any information on the relative efficiencies of para, meta and ortho xylenes in dewaxing paraffin sections for nucleic acid extraction. Does it matter? 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 ********************************************************************** 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 AnthonyH <@t> chw.edu.au Tue May 18 20:08:30 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Efficiency of p o and m xylenes in dewaxing Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E1C8@simba.kids> Hi all, Would anyone have any information on the relative efficiencies of para, meta and ortho xylenes in dewaxing paraffin sections for nucleic acid extraction. Does it matter? 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 ********************************************************************** 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 tmciver <@t> ils-inc.com Wed May 19 04:50:52 2004 From: tmciver <@t> ils-inc.com (Tara Mciver) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] HTL tissue size minimum requriements Message-ID: I attended a readiness workshop in NC and thought I understood the instructor to say that the tissue sizes should be at least the size indicated, so actually the tissue size could be larger, and that is what he suggested we do. Can anybody verify that I understood him correctly? Also, does the tissue have to be a perfect square, or can I just be sure it covers the square representation provided with the test? Thanks for any help! From JWEEMS <@t> sjha.org Wed May 19 06:11:09 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] unsubscribe Message-ID: 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 gliuygao <@t> hotmail.com Wed May 19 08:08:42 2004 From: gliuygao <@t> hotmail.com (yan gao) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] (no subject) Message-ID: Hi, Everyone. I am looking for CD34 antibody stain rat tissues. Anyone knows where I can get it? Thanks. _________________________________________________________________ [1]Express yourself with the new version of MSN Messenger! Download today - it's FREE! References 1. http://g.msn.com/8HMBENUS/2752??PS=47575 From gliuygao <@t> hotmail.com Wed May 19 08:09:28 2004 From: gliuygao <@t> hotmail.com (yan gao) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] (no subject) Message-ID: Hi, Everyone. I am looking for CD34 antibody stain rat tissues. Anyone knows where I can get it? Thanks. Yan Gao Doctoral of Science Novartis _________________________________________________________________ [1]Express yourself with the new version of MSN Messenger! Download today - it's FREE! References 1. http://g.msn.com/8HMBENUS/2752??PS=47575 From gliuygao <@t> hotmail.com Wed May 19 08:05:25 2004 From: gliuygao <@t> hotmail.com (yan gao) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] (no subject) Message-ID: Hi, Everyone. I am looking for CD34 antibody stain rat tissues. Anyone knows where I can get it? Thanks. Yan Gao NOVARTIS _________________________________________________________________ [1]Learn to simplify your finances and your life in Streamline Your Life from MSN Money. References 1. http://g.msn.com/8HMBENUS/2743??PS=47575 From joseph-galbraith <@t> uiowa.edu Wed May 19 08:10:52 2004 From: joseph-galbraith <@t> uiowa.edu (Galbraith, Joe) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] HTL tissue size minimum requriements Message-ID: <5D03ED7B9391D4119D9B0008C76B7B2403005B1F@uihc-mail1.uihc.uiowa.edu> The sizes are minimums as you understand. You will be held strictly accountable for ensuring that the tissue you submit in your block must exceed the dimensions listed and that the tissue on the slide must match the dimensions in the block (without excessive spreading from overfloating on a warm water bath). Remember to allow for shrinkage during processing when you are acquiring your tissue when fixing in formalin and using standard processing. To get 2.0 cm of duodenum into the block you may have to cut 2.5 cm from the source tissue depending on your processing schedule. But 1.9 cm will not due. Also, if they specify 2.0 cm of duodenum with epitelium along one side, they mean complete uninterupted epithelium in perfect condition, perfectly fixed, perfectly processed, perfectly cut, and perfectly stained. Any flaws will cost you points. I don't mean to be negative just to remind everyone preparing for the practical that you must pay attention to all the details. Read the directions very carefully and follow every picky detail that they specify. A significant part of the exam is the applicant's ability to follow written instructions accurately. (Meaning, if they say a square, I'd recommend a square.) Pay attention to the labeling of your slides. Be sure your labels are clear and readable (typed), labeled exactly as instructed, and that your label material is in good condition (stays on the slide, no curls or peeling, etc.) Hope this helps, good luck. Joe Galbraith -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Tara Mciver Sent: Wednesday, May 19, 2004 4:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HTL tissue size minimum requriements I attended a readiness workshop in NC and thought I understood the instructor to say that the tissue sizes should be at least the size indicated, so actually the tissue size could be larger, and that is what he suggested we do. Can anybody verify that I understood him correctly? Also, does the tissue have to be a perfect square, or can I just be sure it covers the square representation provided with the test? Thanks for any help! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pxs76 <@t> po.cwru.edu Wed May 19 08:46:55 2004 From: pxs76 <@t> po.cwru.edu (phyllis) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] (no subject) Message-ID: <5.1.0.14.0.20040519094604.03b9c2e0@pop.cwru.edu> Why are you handling this processing? The CJD Surveillance has been set up by CDC to do all this work for you and also provide a diagnosis. If your pathologists choose to do this themselves, be aware of the protocols which can be found at www.cdc.gov and also the WHO website. The biopsy should be handled in a level 2 facility. You need to wear gloves and face shield. All processing and staining is to been done by hand in individual containers which will be incinirated on completion, otherwise you will have deal with the demanding work of decontaminating the processor. All waste is incinerated. You will find all this info at CDC website. Also refer to www.cjdsurveillance.com if you decide to let the CJD Survellance do this for you. Free of charge. Phyllis Scalzo, HT(ASCP) Case Western Reserve University 2085 Adelbert Rd. Cleveland, Ohio 44106 Ph: 216-368-0822 Fax: 216-368-2546 From convmcm <@t> cc.usu.edu Wed May 19 08:48:52 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] H&E stain problems In-Reply-To: Message-ID: <000a01c43da8$08034a90$4a737b81@Cygnus> Gary Is Gill a relative or yours??? I don't give a hoot who made what when. I like Harris. I don't believe Gill has made any significant improvements over the old dead white guy's hematoxylin. BTW, classical music may have been composed by dead white guys, but I don't hear ANYTHING being composed today that has the depth, complexity and color of those great composers. I've studied piano from since I was 4 yrs old, and I've studied the organ (I've played as a church organist)for almost 20 years. I know music like the back of my hand and I love ALL music (except gangstah stuff). SO DON'T try to tell me about what music is living and worth my while. Dead white Europeans. Yeah, right. I'd like to see someone between 8 and 14 in this generation compose music like Felix Mendelsohnn's or Mozart's. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Tuesday, May 18, 2004 10:04 AM To: 'Connie McManus'; 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems You may have heard that classical music was composed by dead white Europeans. Well, Harris hematoxylin was composed by a dead white American (physician at Jefferson Hospital in 1904). Gill hematoxylin was composed by a live white American. So if you want to liven things up, go with Gill's! Gary Gill (one and the same) PS -- No royalties involved, thanks to bad advice in 1972 from corporate counsel for Johns Hopkins Medical School. -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Tuesday, May 18, 2004 10:41 AM To: 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Wow. What a lot of interesting comments!! I agree with Terry re the agitation. When I watch the stainer do those dips (I can program how many, but NOT the briskness), I wonder if you could even call it agitation. My hand dips are very brisk. Also, I don't bother letting the slides stay in the alcohols for 1 minute or 2, I give the slides about 20 -30 good brisk dips in each solution, then the timed rinses & staining. This has always been far more satisfactory to me than those sllllooooowwwww dips from the machine. As for the kind of hematoxylin, someone suggested I throw out the Harris and do Gills III. I've tried Gill III before and I much prefer the Harris. So it's just a matter of personal preference on that... AND what your pathologist likes *G* Everyone having an nice Tuesday??? *g* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, May 18, 2004 7:35 AM To: Connie McManus; Petia P Stefanova; Megan Kear; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Connie remarks: "In truth, I prefer my hand stained sections better than when they're stained automatically." When I first saw what I call x-y stainers, I thought that we had in this, something that reproduced hand staining. Not so. What I think is lacking is the brisk agitation necessary to break down the interface between old and new solution. I'm not so sure about the rinsing either. 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 18 May 2004 15:09 To: 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ 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 Terry.Marshall <@t> rothgen.nhs.uk Wed May 19 08:56:37 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] H&E stain problems Message-ID: Close relative:-) 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 19 May 2004 14:49 To: 'Gary Gill'; Marshall Terry Dr, Consultant Histopathologist; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Gary Is Gill a relative or yours??? I don't give a hoot who made what when. I like Harris. I don't believe Gill has made any significant improvements over the old dead white guy's hematoxylin. BTW, classical music may have been composed by dead white guys, but I don't hear ANYTHING being composed today that has the depth, complexity and color of those great composers. I've studied piano from since I was 4 yrs old, and I've studied the organ (I've played as a church organist)for almost 20 years. I know music like the back of my hand and I love ALL music (except gangstah stuff). SO DON'T try to tell me about what music is living and worth my while. Dead white Europeans. Yeah, right. I'd like to see someone between 8 and 14 in this generation compose music like Felix Mendelsohnn's or Mozart's. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Tuesday, May 18, 2004 10:04 AM To: 'Connie McManus'; 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems You may have heard that classical music was composed by dead white Europeans. Well, Harris hematoxylin was composed by a dead white American (physician at Jefferson Hospital in 1904). Gill hematoxylin was composed by a live white American. So if you want to liven things up, go with Gill's! Gary Gill (one and the same) PS -- No royalties involved, thanks to bad advice in 1972 from corporate counsel for Johns Hopkins Medical School. -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Tuesday, May 18, 2004 10:41 AM To: 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Wow. What a lot of interesting comments!! I agree with Terry re the agitation. When I watch the stainer do those dips (I can program how many, but NOT the briskness), I wonder if you could even call it agitation. My hand dips are very brisk. Also, I don't bother letting the slides stay in the alcohols for 1 minute or 2, I give the slides about 20 -30 good brisk dips in each solution, then the timed rinses & staining. This has always been far more satisfactory to me than those sllllooooowwwww dips from the machine. As for the kind of hematoxylin, someone suggested I throw out the Harris and do Gills III. I've tried Gill III before and I much prefer the Harris. So it's just a matter of personal preference on that... AND what your pathologist likes *G* Everyone having an nice Tuesday??? *g* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, May 18, 2004 7:35 AM To: Connie McManus; Petia P Stefanova; Megan Kear; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Connie remarks: "In truth, I prefer my hand stained sections better than when they're stained automatically." When I first saw what I call x-y stainers, I thought that we had in this, something that reproduced hand staining. Not so. What I think is lacking is the brisk agitation necessary to break down the interface between old and new solution. I'm not so sure about the rinsing either. 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 18 May 2004 15:09 To: 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ 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 convmcm <@t> cc.usu.edu Wed May 19 09:03:16 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] CJD DECONTAMINATION PROCEDURE In-Reply-To: Message-ID: <000c01c43daa$087de6e0$4a737b81@Cygnus> I don't deal with CJD, but I do a lot of CWD (Chronic Wasting Disease) and Scrapie, which are prion diseases in dear/elk and sheep (respectively). CWD and Scrapie had been associated with humans for hundreds of years with no known transmission of the disease from animal to human. HOWEVER, since prions are not neutralized by fixation of any kind, not killed by autoclaving, this makes them a biohazard in processed tissues. If I worked with CJD, I would ... 1 set up a place that is completely separate from the rest of the lab. Barrier off a section of your lab, use another room (preferable) 2 use disposable bunny suits, shoe covers, face masks, eye wear and DOUBLE GLOVE 3 put stickie mats on the floor completely surrounding the cutting area, 4 put ALL paraffin shavings in a biohaz bag, 5 use paper towels to clean off the microtome and throw those in the biohaz bag 6. incinerate the bunny suit, goggles, gloves, face maskes, etc with the paraffin shavings. 7. Ask for a really big pay raise. Claim it's hazard pay. Have fun. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of srishan@mail.holyname.org Sent: Tuesday, May 18, 2004 9:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CJD DECONTAMINATION PROCEDURE Hello everyone, This question might have been posted before, but, Histology Department is expecting a brain biopsy from a possible CJD patient today. The specimen will be coming down in formalin from the O.R. It will be kept in formalin for 2-7 days and will be transferred to formic acid for the required time. Then the specimen will be transferred to formalin before processing for another 2 days. Has anyone dealt with this procedure before. These are the questions asked by the hstology department. How do you decontaminate the processor, microtome, stainer and coverslipper? What kind of protective gear apart from standard precaution has to be used? How about the shaving of the tissues? How do you dispose the formalin from the original formalin container where the specimen was sent in and how do you dispose the reagents from the processor and the stainer? Can this biopsy be processed, once treated with formic acid, along with the other specimens or should the biopsy be processed with the other specimens? Need Help!! Thank you in advance Mala Srishan Histology Supervisor Holy Name Hospital Teaneck, NJ 07666 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BWilliams <@t> Lifespan.org Wed May 19 09:22:53 2004 From: BWilliams <@t> Lifespan.org (Williams, Blanche) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] help Message-ID: <85151E3A0A49DF498F39F19EE56FEA8D0317CECF@lsexch2.lsmaster.lifespan.org> Can someone answer a question regarding slide retention for me? How long to labs keep their immunofluorescence slides? -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Wednesday, May 19, 2004 10:16 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 6, Issue 28 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. CJD DECONTAMINATION PROCEDURE (srishan@mail.holyname.org) 2. RE: H&E stain problems (Gary Gill) 3. RE: CJD DECONTAMINATION PROCEDURE (Bartlett, Jeanine) 4. RE: CJD DECONTAMINATION PROCEDURE (Kari Bradshaw) 5. research charges (Diane M Nelson) 6. RE: CJD DECONTAMINATION PROCEDURE (Monson, Frederick ) 7. Presto Fix - ? (Richard Cartun) 8. Re: Aussies and Kiwis standing up at microtomes (P. Emry) 9. Congo Red (Karen Wittler) 10. Re: MIcrotome alignment (Ford Royer) 11. RE: research charges (Mass Histology Service) 12. Efficiency of p o and m xylenes in dewaxing (Tony Henwood) 13. Efficiency of p o and m xylenes in dewaxing (Tony Henwood) 14. HTL tissue size minimum requriements (Tara Mciver) 15. unsubscribe (Weems, Joyce) 16. (no subject) (yan gao) 17. (no subject) (yan gao) 18. (no subject) (yan gao) 19. RE: HTL tissue size minimum requriements (Galbraith, Joe) 20. (no subject) (phyllis) 21. RE: H&E stain problems (Connie McManus) 22. RE: H&E stain problems (Marshall Terry Dr, Consultant Histopathologist) 23. RE: CJD DECONTAMINATION PROCEDURE (Connie McManus) ---------------------------------------------------------------------- Message: 1 Date: Tue, 18 May 2004 12:58:33 -0400 From: srishan@mail.holyname.org Subject: [Histonet] CJD DECONTAMINATION PROCEDURE To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Hello everyone, This question might have been posted before, but, Histology Department is expecting a brain biopsy from a possible CJD patient today. The specimen will be coming down in formalin from the O.R. It will be kept in formalin for 2-7 days and will be transferred to formic acid for the required time. Then the specimen will be transferred to formalin before processing for another 2 days. Has anyone dealt with this procedure before. These are the questions asked by the hstology department. How do you decontaminate the processor, microtome, stainer and coverslipper? What kind of protective gear apart from standard precaution has to be used? How about the shaving of the tissues? How do you dispose the formalin from the original formalin container where the specimen was sent in and how do you dispose the reagents from the processor and the stainer? Can this biopsy be processed, once treated with formic acid, along with the other specimens or should the biopsy be processed with the other specimens? Need Help!! Thank you in advance Mala Srishan Histology Supervisor Holy Name Hospital Teaneck, NJ 07666 ------------------------------ Message: 2 Date: Tue, 18 May 2004 12:03:43 -0500 From: Gary Gill Subject: RE: [Histonet] H&E stain problems To: 'Connie McManus' , "'Marshall Terry Dr,Consultant Histopathologist'" , 'Petia P Stefanova' , 'Megan Kear' , histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain You may have heard that classical music was composed by dead white Europeans. Well, Harris hematoxylin was composed by a dead white American (physician at Jefferson Hospital in 1904). Gill hematoxylin was composed by a live white American. So if you want to liven things up, go with Gill's! Gary Gill (one and the same) PS -- No royalties involved, thanks to bad advice in 1972 from corporate counsel for Johns Hopkins Medical School. -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Tuesday, May 18, 2004 10:41 AM To: 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Wow. What a lot of interesting comments!! I agree with Terry re the agitation. When I watch the stainer do those dips (I can program how many, but NOT the briskness), I wonder if you could even call it agitation. My hand dips are very brisk. Also, I don't bother letting the slides stay in the alcohols for 1 minute or 2, I give the slides about 20 -30 good brisk dips in each solution, then the timed rinses & staining. This has always been far more satisfactory to me than those sllllooooowwwww dips from the machine. As for the kind of hematoxylin, someone suggested I throw out the Harris and do Gills III. I've tried Gill III before and I much prefer the Harris. So it's just a matter of personal preference on that... AND what your pathologist likes *G* Everyone having an nice Tuesday??? *g* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, May 18, 2004 7:35 AM To: Connie McManus; Petia P Stefanova; Megan Kear; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Connie remarks: "In truth, I prefer my hand stained sections better than when they're stained automatically." When I first saw what I call x-y stainers, I thought that we had in this, something that reproduced hand staining. Not so. What I think is lacking is the brisk agitation necessary to break down the interface between old and new solution. I'm not so sure about the rinsing either. 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 18 May 2004 15:09 To: 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ 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 ------------------------------ Message: 3 Date: Tue, 18 May 2004 13:58:07 -0400 From: "Bartlett, Jeanine" Subject: RE: [Histonet] CJD DECONTAMINATION PROCEDURE To: , Message-ID: Content-Type: text/plain; charset="us-ascii" The WHO booklet was too large to send as an attachment so it couldn't be delivered. I will try and send you what I think is the most relevant chapter. Perhaps you could visit the WHO site and get more information if needed. In the meantime, the Surveillance Center link below should have enough information. Jeanine - Mala: There is a lot of conflicting information out there but here is information from 2 very credible sources for you to review. One is the National Surveillance Center at case Western and the other is the WHO. Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 http://www.cjdsurveillance.com and the WHO booklet is sent as an attachment. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of srishan@mail.holyname.org Sent: Tuesday, May 18, 2004 12:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CJD DECONTAMINATION PROCEDURE Hello everyone, This question might have been posted before, but, Histology Department is expecting a brain biopsy from a possible CJD patient today. The specimen will be coming down in formalin from the O.R. It will be kept in formalin for 2-7 days and will be transferred to formic acid for the required time. Then the specimen will be transferred to formalin before processing for another 2 days. Has anyone dealt with this procedure before. These are the questions asked by the hstology department. How do you decontaminate the processor, microtome, stainer and coverslipper? What kind of protective gear apart from standard precaution has to be used? How about the shaving of the tissues? How do you dispose the formalin from the original formalin container where the specimen was sent in and how do you dispose the reagents from the processor and the stainer? Can this biopsy be processed, once treated with formic acid, along with the other specimens or should the biopsy be processed with the other specimens? Need Help!! Thank you in advance Mala Srishan Histology Supervisor Holy Name Hospital Teaneck, NJ 07666 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Tue, 18 May 2004 10:55:17 -0700 From: "Kari Bradshaw" Subject: RE: [Histonet] CJD DECONTAMINATION PROCEDURE To: , Message-ID: Content-Type: text/plain; charset="US-ASCII" These people have all the answers: National Prion Pathology Surveillance Center Case Western Reserve University 2085 Adelbert Road Cleveland,Ohio 44106 216-368-0587 www.cjdsurveillance.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of srishan@mail.holyname.org Sent: Tuesday, May 18, 2004 9:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CJD DECONTAMINATION PROCEDURE Hello everyone, This question might have been posted before, but, Histology Department is expecting a brain biopsy from a possible CJD patient today. The specimen will be coming down in formalin from the O.R. It will be kept in formalin for 2-7 days and will be transferred to formic acid for the required time. Then the specimen will be transferred to formalin before processing for another 2 days. Has anyone dealt with this procedure before. These are the questions asked by the hstology department. How do you decontaminate the processor, microtome, stainer and coverslipper? What kind of protective gear apart from standard precaution has to be used? How about the shaving of the tissues? How do you dispose the formalin from the original formalin container where the specimen was sent in and how do you dispose the reagents from the processor and the stainer? Can this biopsy be processed, once treated with formic acid, along with the other specimens or should the biopsy be processed with the other specimens? Need Help!! Thank you in advance Mala Srishan Histology Supervisor Holy Name Hospital Teaneck, NJ 07666 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Tue, 18 May 2004 13:03:49 -0500 From: Diane M Nelson Subject: [Histonet] research charges To: histonet@lists.utsouthwestern.edu Message-ID: <6.1.0.6.2.20040518124918.01b5caa0@dmnelson.mail.iastate.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Hi Histonetters! Our veterinary lab is in the process of up dating our research charges. I would like to know what other labs/hospitals charge for H & E bone slides and H & E eye slides. Currently we are charging, Bone: $19.00 for decalcifying in 25% formic acid, process, embed, sectioning and H & E stain. We are losing money on our larger bone projects. Our charges for processing, embedding sectioning and H & E stain for eye slides is $9.00. It also depends on the project rather or not we make or lose money. The researchers do their own grossing of tissue. Any help would be greatly appreciated! Thanks in advance. Diane Gerjets Iowa State University ------------------------------ Message: 6 Date: Tue, 18 May 2004 14:04:45 -0400 From: "Monson, Frederick " Subject: RE: [Histonet] CJD DECONTAMINATION PROCEDURE To: Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" http://www.nursingceu.com/NCEU/courses/prions/ http://www.mad-cow.org/tonsil_test2.html#Biosafety A start. Cheers Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone/FAX: 610-738-0437 eMail: fmonson@wcupa.edu CASI Page and Scheduling http://darwin.wcupa.edu/CASI/ -----Original Message----- From: srishan@mail.holyname.org [mailto:srishan@mail.holyname.org] Sent: Tuesday, May 18, 2004 12:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CJD DECONTAMINATION PROCEDURE Hello everyone, This question might have been posted before, but, Histology Department is expecting a brain biopsy from a possible CJD patient today. The specimen will be coming down in formalin from the O.R. It will be kept in formalin for 2-7 days and will be transferred to formic acid for the required time. Then the specimen will be transferred to formalin before processing for another 2 days. Has anyone dealt with this procedure before. These are the questions asked by the hstology department. How do you decontaminate the processor, microtome, stainer and coverslipper? What kind of protective gear apart from standard precaution has to be used? How about the shaving of the tissues? How do you dispose the formalin from the original formalin container where the specimen was sent in and how do you dispose the reagents from the processor and the stainer? Can this biopsy be processed, once treated with formic acid, along with the other specimens or should the biopsy be processed with the other specimens? Need Help!! Thank you in advance Mala Srishan Histology Supervisor Holy Name Hospital Teaneck, NJ 07666 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Tue, 18 May 2004 14:32:48 -0400 From: "Richard Cartun" Subject: [Histonet] Presto Fix - ? To: Message-ID: Content-Type: text/plain; charset=US-ASCII Anyone using "Presto Fix" for bone marrow core biopsies? If so, what does it do to immunoreactivity? Thank you. Richard Cartun ------------------------------ Message: 8 Date: Tue, 18 May 2004 11:34:18 -0700 (PDT) From: "P. Emry" Subject: Re: [Histonet] Aussies and Kiwis standing up at microtomes To: Andrew Kennedy Cc: Histonet Message-ID: Content-Type: TEXT/PLAIN; charset=US-ASCII Ahhhhh working with a beer at my elbow. Sounds very sane to me. I think we got one of your country women..a Wealthal..up for a phd. She denies dwarf tossing, but I have heard things to the contrary. I have two square carts with drawers in used in the dental school. They have wheels on them. I put the microtome on one and the waterbath on the other. They sit side-by side with a space between. They are just high enough for me to use a standard height chair with wheels and move back and forth between them. It is the best arrangement I have found. I can get up over the water bath to get a good look at what is going on. I really like it. Not hard on the back like benches. Would suggest it for those having problems with benches and sitting still in one possition. If I can figure out how to put an ice bucket for the beer between, we's really have something. Trisha On Mon, 17 May 2004, Andrew Kennedy wrote: > Hi Tim and Fred (and the rest of Histonetters too!!) > > > > In my experience, we Aussies don't like to stand up at the microtome if we > can help it. Standing and cutting sections makes no sense at all - we might > spill our beer and not be able to see the television that sits on the bench > next to us so we can watch Aussie Rules (I prefer Rugby League, by the way) > > :-) > > But seriously folks, you would have to have high benches and even then you > would probably hunch over the microtome - I remember seeing a histotech > cutting standing up years ago when I was a lab assistant. She turned the > microtome sideways, stood at the bench (which was desk height) and stooped > over the microtome. That can't be good for your back! It was the most > uncomfortable looking cutting position I've ever seen. > > > > Overlapping with the safety thread that is currently quite "hot", we really > owe it to ourselves and to those we work with to be safe all the time! > > > > In Australia, Occupational Health and Safety is an important issue that > affects everything we do in the workplace. However, any safety procedure > that has been implemented in our labs is a result of consultation with the > parties at stake. Safe Operating Procedures are designed by the people who > will use them, not by some beaurocrat sitting in an office telling others > what to do! Take control of your safety and use common sense at the same > time! Makes sense to me!! > > > > Andrew Kennedy > > Senior Science Officer > > Anatomical Pathology > > Concord Repatriation General Hospital > > Hospital Road > Concord NSW Australia 2139 > > > > ph: +612 9767 6115 > > Fax +612 9767 8427 > > > > "oderint dum metuant" > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 9 Date: Tue, 18 May 2004 16:29:47 -0400 From: Karen Wittler Subject: [Histonet] Congo Red To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII We stain our Congo Reds on an automated stainer now. Before that we stained with hematoxylin first because rinsing and differentiation was required and we used an alkaline Congo Red. After staining with the Congo Red in a saturated sodium chloride- alcohol solution,we did not rinse,but rapidly dehydrated, cleared and mounted For our H&Es we use Modified Harris Hematoxylin and Eosin-Y w/ Phloxine both manufactured by Richard Allen. Karen Wittler, HT(ASCP) Johns Hopkins Hospital Baltimore, MD ------------------------------ Message: 10 Date: Tue, 18 May 2004 16:16:32 -0500 From: "Ford Royer" Subject: Re: [Histonet] MIcrotome alignment To: KHays@mbhs.org, histonet@lists.utsouthwestern.edu Cc: Marcia Welch Message-ID: <40AA7D30.10601@bitstream.net> Content-Type: text/plain; charset=us-ascii; format=flowed On their web site catalog, they show a price of $775.00. Contact Marcia Welch at: 800-383-7799 or Email: for a customer price quote. Ford M. Royer, MT(ASCP) Analytical Instruments, llc 1200 Mendelssohn Ave. N., Ste. 50 Minneapolis, MN 55427 (800) 565-1895, ext. 17 URL: KHays@mbhs.org wrote: > > Kathy Tedford-Hays HT (ASCP) > Technical Specialist, Histology Dept > (601)-968-3070 ext 7398 > Baptist Medical Center > 1225 North State Street > Jackson, MS 39202 > > how much is it , do you know? > > > > "Ford Royer" > Sent by: histonet-bounces@lists.utsouthwestern.edu > > 05/18/2004 09:00 AM > > > To: histonet@lists.utsouthwestern.edu > cc: > Fax to: > Subject: Re: [Histonet] MIcrotome alignment > > > > > Newcomer Supply, Madison, WI sells a microtome aligner. Product Code > 6255A > Web URL > Check it out. > > ~ Ford > Ford M. Royer, MT(ASCP) > Analytical Instruments, llc > Minneapolis, MN 55427 > (800) 565-1895, Ext. 17 > > > Roberts, Jon wrote: > > >I have recently purchased a Leica RM2255 with the new precise > specimen orientation system that adjust to an exact zero position or > x-y axis standard. But I also have old leicas without this system. > Is there any quick and easy way to align my old leicas to the new > one???? Just wondering? I though I've heard of such a device, but > wonder if anyone else has heard of it or actually used it? > > > > > >CONFIDENTIALITY NOTICE: This email message and any accompanying data > are confidential, and intended only for the named recipient(s). If > you are not the intended recipient(s), you are hereby notified that > the dissemination, distribution, and or copying of this message is > strictly prohibited. If you receive this message in error, or are not > the named recipient(s), please notify the sender at the email address > above, delete this email from your computer, and destroy any copies in > any form 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 > This internet email has been certified virus free by BHS Anti-Virus > system. > > ------------------------------ Message: 11 Date: Tue, 18 May 2004 19:59:45 -0400 From: "Mass Histology Service" Subject: RE: [Histonet] research charges To: "Diane M Nelson" , Message-ID: Content-Type: text/plain; charset="us-ascii" Diane, We charge one price across the board, $5.50 if the tissue is received in a cassette and $2.50 if it needs decalcification. Our philosophy is that we don't charge less for simple specimens, so why charge more for occasional complicated ones? Of course if it's a large project consisting of many whole eyes or femur heads, I'd probably double our current prices to cover for the additional labor and material. Jim ____________________________ James E. Staruk, HT(ASCP) Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Diane M Nelson Sent: Tuesday, May 18, 2004 2:04 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] research charges Hi Histonetters! Our veterinary lab is in the process of up dating our research charges. I would like to know what other labs/hospitals charge for H & E bone slides and H & E eye slides. Currently we are charging, Bone: $19.00 for decalcifying in 25% formic acid, process, embed, sectioning and H & E stain. We are losing money on our larger bone projects. Our charges for processing, embedding sectioning and H & E stain for eye slides is $9.00. It also depends on the project rather or not we make or lose money. The researchers do their own grossing of tissue. Any help would be greatly appreciated! Thanks in advance. Diane Gerjets Iowa State University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Wed, 19 May 2004 11:08:30 +1000 From: Tony Henwood Subject: [Histonet] Efficiency of p o and m xylenes in dewaxing To: "Histonet (Histonet@lists.utsouthwestern.edu)" , "'histonet@pathology.swmed.edu'" Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E1C8@simba.kids> Content-Type: text/plain Hi all, Would anyone have any information on the relative efficiencies of para, meta and ortho xylenes in dewaxing paraffin sections for nucleic acid extraction. Does it matter? 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 ********************************************************************** 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. ********************************************************************** ------------------------------ Message: 13 Date: Wed, 19 May 2004 11:08:30 +1000 From: Tony Henwood Subject: [Histonet] Efficiency of p o and m xylenes in dewaxing To: "Histonet (Histonet@lists.utsouthwestern.edu)" , "'histonet@pathology.swmed.edu'" Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E1C8@simba.kids> Content-Type: text/plain Hi all, Would anyone have any information on the relative efficiencies of para, meta and ortho xylenes in dewaxing paraffin sections for nucleic acid extraction. Does it matter? 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 ********************************************************************** 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. ********************************************************************** ------------------------------ Message: 14 Date: Wed, 19 May 2004 05:50:52 -0400 From: "Tara Mciver" Subject: [Histonet] HTL tissue size minimum requriements To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" I attended a readiness workshop in NC and thought I understood the instructor to say that the tissue sizes should be at least the size indicated, so actually the tissue size could be larger, and that is what he suggested we do. Can anybody verify that I understood him correctly? Also, does the tissue have to be a perfect square, or can I just be sure it covers the square representation provided with the test? Thanks for any help! ------------------------------ Message: 15 Date: Wed, 19 May 2004 07:11:09 -0400 From: "Weems, Joyce" Subject: [Histonet] unsubscribe To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" 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. ------------------------------ Message: 16 Date: Wed, 19 May 2004 09:08:42 -0400 From: "yan gao" Subject: [Histonet] (no subject) To: histonet@lists.utsouthwestern.edu Cc: gliuygao@hotmail.com Message-ID: Content-Type: text/plain Hi, Everyone. I am looking for CD34 antibody stain rat tissues. Anyone knows where I can get it? Thanks. _________________________________________________________________ [1]Express yourself with the new version of MSN Messenger! Download today - it's FREE! References 1. http://g.msn.com/8HMBENUS/2752??PS=47575 ------------------------------ Message: 17 Date: Wed, 19 May 2004 09:09:28 -0400 From: "yan gao" Subject: [Histonet] (no subject) To: histonet@lists.utsouthwestern.edu Cc: gliuygao@hotmail.com Message-ID: Content-Type: text/plain Hi, Everyone. I am looking for CD34 antibody stain rat tissues. Anyone knows where I can get it? Thanks. Yan Gao Doctoral of Science Novartis _________________________________________________________________ [1]Express yourself with the new version of MSN Messenger! Download today - it's FREE! References 1. http://g.msn.com/8HMBENUS/2752??PS=47575 ------------------------------ Message: 18 Date: Wed, 19 May 2004 09:05:25 -0400 From: "yan gao" Subject: [Histonet] (no subject) To: histonet@list.utsouthwestern.edu, histonet@pathology.swmed.edu Cc: gliuygao@hotmail.com Message-ID: Content-Type: text/plain Hi, Everyone. I am looking for CD34 antibody stain rat tissues. Anyone knows where I can get it? Thanks. Yan Gao NOVARTIS _________________________________________________________________ [1]Learn to simplify your finances and your life in Streamline Your Life from MSN Money. References 1. http://g.msn.com/8HMBENUS/2743??PS=47575 ------------------------------ Message: 19 Date: Wed, 19 May 2004 08:10:52 -0500 From: "Galbraith, Joe" Subject: RE: [Histonet] HTL tissue size minimum requriements To: "Tara Mciver" , Message-ID: <5D03ED7B9391D4119D9B0008C76B7B2403005B1F@uihc-mail1.uihc.uiowa.edu> Content-Type: text/plain; charset="iso-8859-1" The sizes are minimums as you understand. You will be held strictly accountable for ensuring that the tissue you submit in your block must exceed the dimensions listed and that the tissue on the slide must match the dimensions in the block (without excessive spreading from overfloating on a warm water bath). Remember to allow for shrinkage during processing when you are acquiring your tissue when fixing in formalin and using standard processing. To get 2.0 cm of duodenum into the block you may have to cut 2.5 cm from the source tissue depending on your processing schedule. But 1.9 cm will not due. Also, if they specify 2.0 cm of duodenum with epitelium along one side, they mean complete uninterupted epithelium in perfect condition, perfectly fixed, perfectly processed, perfectly cut, and perfectly stained. Any flaws will cost you points. I don't mean to be negative just to remind everyone preparing for the practical that you must pay attention to all the details. Read the directions very carefully and follow every picky detail that they specify. A significant part of the exam is the applicant's ability to follow written instructions accurately. (Meaning, if they say a square, I'd recommend a square.) Pay attention to the labeling of your slides. Be sure your labels are clear and readable (typed), labeled exactly as instructed, and that your label material is in good condition (stays on the slide, no curls or peeling, etc.) Hope this helps, good luck. Joe Galbraith -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Tara Mciver Sent: Wednesday, May 19, 2004 4:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HTL tissue size minimum requriements I attended a readiness workshop in NC and thought I understood the instructor to say that the tissue sizes should be at least the size indicated, so actually the tissue size could be larger, and that is what he suggested we do. Can anybody verify that I understood him correctly? Also, does the tissue have to be a perfect square, or can I just be sure it covers the square representation provided with the test? Thanks for any help! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 20 Date: Wed, 19 May 2004 09:46:55 -0400 From: phyllis Subject: [Histonet] (no subject) To: histonet@lists.utsouthwestern.edu Message-ID: <5.1.0.14.0.20040519094604.03b9c2e0@pop.cwru.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Why are you handling this processing? The CJD Surveillance has been set up by CDC to do all this work for you and also provide a diagnosis. If your pathologists choose to do this themselves, be aware of the protocols which can be found at www.cdc.gov and also the WHO website. The biopsy should be handled in a level 2 facility. You need to wear gloves and face shield. All processing and staining is to been done by hand in individual containers which will be incinirated on completion, otherwise you will have deal with the demanding work of decontaminating the processor. All waste is incinerated. You will find all this info at CDC website. Also refer to www.cjdsurveillance.com if you decide to let the CJD Survellance do this for you. Free of charge. Phyllis Scalzo, HT(ASCP) Case Western Reserve University 2085 Adelbert Rd. Cleveland, Ohio 44106 Ph: 216-368-0822 Fax: 216-368-2546 ------------------------------ Message: 21 Date: Wed, 19 May 2004 07:48:52 -0600 From: "Connie McManus" Subject: RE: [Histonet] H&E stain problems To: "'Gary Gill'" , "'Marshall Terry Dr, Consultant Histopathologist'" , "'Petia P Stefanova'" , "'Megan Kear'" , Cc: histonet@lists.utsouthwestern.edu Message-ID: <000a01c43da8$08034a90$4a737b81@Cygnus> Content-Type: text/plain; charset="us-ascii" Gary Is Gill a relative or yours??? I don't give a hoot who made what when. I like Harris. I don't believe Gill has made any significant improvements over the old dead white guy's hematoxylin. BTW, classical music may have been composed by dead white guys, but I don't hear ANYTHING being composed today that has the depth, complexity and color of those great composers. I've studied piano from since I was 4 yrs old, and I've studied the organ (I've played as a church organist)for almost 20 years. I know music like the back of my hand and I love ALL music (except gangstah stuff). SO DON'T try to tell me about what music is living and worth my while. Dead white Europeans. Yeah, right. I'd like to see someone between 8 and 14 in this generation compose music like Felix Mendelsohnn's or Mozart's. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Tuesday, May 18, 2004 10:04 AM To: 'Connie McManus'; 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems You may have heard that classical music was composed by dead white Europeans. Well, Harris hematoxylin was composed by a dead white American (physician at Jefferson Hospital in 1904). Gill hematoxylin was composed by a live white American. So if you want to liven things up, go with Gill's! Gary Gill (one and the same) PS -- No royalties involved, thanks to bad advice in 1972 from corporate counsel for Johns Hopkins Medical School. -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Tuesday, May 18, 2004 10:41 AM To: 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Wow. What a lot of interesting comments!! I agree with Terry re the agitation. When I watch the stainer do those dips (I can program how many, but NOT the briskness), I wonder if you could even call it agitation. My hand dips are very brisk. Also, I don't bother letting the slides stay in the alcohols for 1 minute or 2, I give the slides about 20 -30 good brisk dips in each solution, then the timed rinses & staining. This has always been far more satisfactory to me than those sllllooooowwwww dips from the machine. As for the kind of hematoxylin, someone suggested I throw out the Harris and do Gills III. I've tried Gill III before and I much prefer the Harris. So it's just a matter of personal preference on that... AND what your pathologist likes *G* Everyone having an nice Tuesday??? *g* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, May 18, 2004 7:35 AM To: Connie McManus; Petia P Stefanova; Megan Kear; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Connie remarks: "In truth, I prefer my hand stained sections better than when they're stained automatically." When I first saw what I call x-y stainers, I thought that we had in this, something that reproduced hand staining. Not so. What I think is lacking is the brisk agitation necessary to break down the interface between old and new solution. I'm not so sure about the rinsing either. 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 18 May 2004 15:09 To: 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ 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 ------------------------------ Message: 22 Date: Wed, 19 May 2004 14:56:37 +0100 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] H&E stain problems To: "Connie McManus" , "Gary Gill" , "Petia P Stefanova" , "Megan Kear" , Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" Close relative:-) 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 19 May 2004 14:49 To: 'Gary Gill'; Marshall Terry Dr, Consultant Histopathologist; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Gary Is Gill a relative or yours??? I don't give a hoot who made what when. I like Harris. I don't believe Gill has made any significant improvements over the old dead white guy's hematoxylin. BTW, classical music may have been composed by dead white guys, but I don't hear ANYTHING being composed today that has the depth, complexity and color of those great composers. I've studied piano from since I was 4 yrs old, and I've studied the organ (I've played as a church organist)for almost 20 years. I know music like the back of my hand and I love ALL music (except gangstah stuff). SO DON'T try to tell me about what music is living and worth my while. Dead white Europeans. Yeah, right. I'd like to see someone between 8 and 14 in this generation compose music like Felix Mendelsohnn's or Mozart's. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Tuesday, May 18, 2004 10:04 AM To: 'Connie McManus'; 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems You may have heard that classical music was composed by dead white Europeans. Well, Harris hematoxylin was composed by a dead white American (physician at Jefferson Hospital in 1904). Gill hematoxylin was composed by a live white American. So if you want to liven things up, go with Gill's! Gary Gill (one and the same) PS -- No royalties involved, thanks to bad advice in 1972 from corporate counsel for Johns Hopkins Medical School. -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Tuesday, May 18, 2004 10:41 AM To: 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Wow. What a lot of interesting comments!! I agree with Terry re the agitation. When I watch the stainer do those dips (I can program how many, but NOT the briskness), I wonder if you could even call it agitation. My hand dips are very brisk. Also, I don't bother letting the slides stay in the alcohols for 1 minute or 2, I give the slides about 20 -30 good brisk dips in each solution, then the timed rinses & staining. This has always been far more satisfactory to me than those sllllooooowwwww dips from the machine. As for the kind of hematoxylin, someone suggested I throw out the Harris and do Gills III. I've tried Gill III before and I much prefer the Harris. So it's just a matter of personal preference on that... AND what your pathologist likes *G* Everyone having an nice Tuesday??? *g* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, May 18, 2004 7:35 AM To: Connie McManus; Petia P Stefanova; Megan Kear; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Connie remarks: "In truth, I prefer my hand stained sections better than when they're stained automatically." When I first saw what I call x-y stainers, I thought that we had in this, something that reproduced hand staining. Not so. What I think is lacking is the brisk agitation necessary to break down the interface between old and new solution. I'm not so sure about the rinsing either. 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 18 May 2004 15:09 To: 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ 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 ------------------------------ Message: 23 Date: Wed, 19 May 2004 08:03:16 -0600 From: "Connie McManus" Subject: RE: [Histonet] CJD DECONTAMINATION PROCEDURE To: , Message-ID: <000c01c43daa$087de6e0$4a737b81@Cygnus> Content-Type: text/plain; charset="us-ascii" I don't deal with CJD, but I do a lot of CWD (Chronic Wasting Disease) and Scrapie, which are prion diseases in dear/elk and sheep (respectively). CWD and Scrapie had been associated with humans for hundreds of years with no known transmission of the disease from animal to human. HOWEVER, since prions are not neutralized by fixation of any kind, not killed by autoclaving, this makes them a biohazard in processed tissues. If I worked with CJD, I would ... 1 set up a place that is completely separate from the rest of the lab. Barrier off a section of your lab, use another room (preferable) 2 use disposable bunny suits, shoe covers, face masks, eye wear and DOUBLE GLOVE 3 put stickie mats on the floor completely surrounding the cutting area, 4 put ALL paraffin shavings in a biohaz bag, 5 use paper towels to clean off the microtome and throw those in the biohaz bag 6. incinerate the bunny suit, goggles, gloves, face maskes, etc with the paraffin shavings. 7. Ask for a really big pay raise. Claim it's hazard pay. Have fun. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of srishan@mail.holyname.org Sent: Tuesday, May 18, 2004 9:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CJD DECONTAMINATION PROCEDURE Hello everyone, This question might have been posted before, but, Histology Department is expecting a brain biopsy from a possible CJD patient today. The specimen will be coming down in formalin from the O.R. It will be kept in formalin for 2-7 days and will be transferred to formic acid for the required time. Then the specimen will be transferred to formalin before processing for another 2 days. Has anyone dealt with this procedure before. These are the questions asked by the hstology department. How do you decontaminate the processor, microtome, stainer and coverslipper? What kind of protective gear apart from standard precaution has to be used? How about the shaving of the tissues? How do you dispose the formalin from the original formalin container where the specimen was sent in and how do you dispose the reagents from the processor and the stainer? Can this biopsy be processed, once treated with formic acid, along with the other specimens or should the biopsy be processed with the other specimens? Need Help!! Thank you in advance Mala Srishan Histology Supervisor Holy Name Hospital Teaneck, NJ 07666 _______________________________________________ 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 End of Histonet Digest, Vol 6, Issue 28 *************************************** From ESmith <@t> CuraGen.com Wed May 19 09:42:16 2004 From: ESmith <@t> CuraGen.com (Smith, Emily) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Background Staining Message-ID: Hello All, In the course of a recent experiment, I saw strong background staining on charged slides with frozen cells in OCT (optimal cutting temperature) when treated with streptavidin-HRP. The staining appears over the entire glass surface not only where the cells and OCT were. This all-over-staining also appears on clean (without OCT on them) slides that are processed with the OCT slides. I have tried extensive water washes and the results remain the same. No primary antibody is added and the results are the same. Have you seen this before? Do you have any suggestions? Thank you for your help. ~Emily Emily A. Smith CuraGen Corporation 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 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 Barry.R.Rittman <@t> uth.tmc.edu Wed May 19 09:55:48 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Slide storage Message-ID: <566FB0B522443D43AF02D2ADBE35A6F077FE0B@UTHEVS3.mail.uthouston.edu> Blanche This may be somewhat of a mute point as many of the fluorescent markers fade over time, sometimes over very short times. This is especially true when exposed to the viewing illumination for some time. It also depends on the storage conditions, are these slides frozen, paraffin, what mountant if any are they in and what conditions of storage? It would be best if a record is required to photograph the image on which the diagnosis is made and store this image. I realize that for many labs this may not be a possibility. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Williams, Blanche Sent: Wednesday, May 19, 2004 9:23 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] help Can someone answer a question regarding slide retention for me? How long to labs keep their immunofluorescence slides? From mab70 <@t> medschl.cam.ac.uk Wed May 19 09:49:09 2004 From: mab70 <@t> medschl.cam.ac.uk (Margaret Blount) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] H&E stain problems Message-ID: <6C21D947F7189448AB447C57BF2992B4025F16C4@mius.medlan.cam.ac.uk> Dear Connie, Hey, don't take it all so seriously, we all tend to generalise sometimes! It's great to hear about folks' outside interests and I congratulate you on being an accomplished musician. I tend to aggree with you about Mendelsohnn and Mozart, but as a limey, I also like Elgar who is one of my heroes. By the way, I use Gill's No 3. Have a good wednesday. I shall be going home to do some gardening in just over an hour - one of my hobbies. Regards margaret Margaret Blount Chief Technician Clinical Biochemistry University of Cambridge Addenbrooke's Hospital Hills Road Cambridge CB2 2QR -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Connie McManus Sent: Wednesday, May 19, 2004 2:49 PM To: 'Gary Gill'; 'Marshall Terry Dr, Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Gary Is Gill a relative or yours??? I don't give a hoot who made what when. I like Harris. I don't believe Gill has made any significant improvements over the old dead white guy's hematoxylin. BTW, classical music may have been composed by dead white guys, but I don't hear ANYTHING being composed today that has the depth, complexity and color of those great composers. I've studied piano from since I was 4 yrs old, and I've studied the organ (I've played as a church organist)for almost 20 years. I know music like the back of my hand and I love ALL music (except gangstah stuff). SO DON'T try to tell me about what music is living and worth my while. Dead white Europeans. Yeah, right. I'd like to see someone between 8 and 14 in this generation compose music like Felix Mendelsohnn's or Mozart's. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Tuesday, May 18, 2004 10:04 AM To: 'Connie McManus'; 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems You may have heard that classical music was composed by dead white Europeans. Well, Harris hematoxylin was composed by a dead white American (physician at Jefferson Hospital in 1904). Gill hematoxylin was composed by a live white American. So if you want to liven things up, go with Gill's! Gary Gill (one and the same) PS -- No royalties involved, thanks to bad advice in 1972 from corporate counsel for Johns Hopkins Medical School. -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Tuesday, May 18, 2004 10:41 AM To: 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Wow. What a lot of interesting comments!! I agree with Terry re the agitation. When I watch the stainer do those dips (I can program how many, but NOT the briskness), I wonder if you could even call it agitation. My hand dips are very brisk. Also, I don't bother letting the slides stay in the alcohols for 1 minute or 2, I give the slides about 20 -30 good brisk dips in each solution, then the timed rinses & staining. This has always been far more satisfactory to me than those sllllooooowwwww dips from the machine. As for the kind of hematoxylin, someone suggested I throw out the Harris and do Gills III. I've tried Gill III before and I much prefer the Harris. So it's just a matter of personal preference on that... AND what your pathologist likes *G* Everyone having an nice Tuesday??? *g* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, May 18, 2004 7:35 AM To: Connie McManus; Petia P Stefanova; Megan Kear; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Connie remarks: "In truth, I prefer my hand stained sections better than when they're stained automatically." When I first saw what I call x-y stainers, I thought that we had in this, something that reproduced hand staining. Not so. What I think is lacking is the brisk agitation necessary to break down the interface between old and new solution. I'm not so sure about the rinsing either. 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 18 May 2004 15:09 To: 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ 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 mab70 <@t> medschl.cam.ac.uk Wed May 19 09:58:12 2004 From: mab70 <@t> medschl.cam.ac.uk (Margaret Blount) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Background Staining Message-ID: <6C21D947F7189448AB447C57BF2992B4025F16C5@mius.medlan.cam.ac.uk> Dear Emily, That's a strange one. Have you titrated your secondary antibody sufficiently? what blocking solution do you use? Have the slides inadvertently dried out at any stage of the procedure? These are a few thoughts that spring to mind. The other thing that I would look at would be to try charged slides from another supplier, or try Polysine or APES slides for comparison on indeed another batch of slides. That's my 2 penny worth - maybe someone else will have something more helpful to say and no doubt you have thought of some or all of these ideas yourself. Good luck. Regards Margaret Margaret Blount Chief Technician Clinical Biochemistry University of Cambridge Addenbrooke's Hospital Hills Road Cambridge CB2 2QR -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Smith, Emily Sent: Wednesday, May 19, 2004 3:42 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Background Staining Hello All, In the course of a recent experiment, I saw strong background staining on charged slides with frozen cells in OCT (optimal cutting temperature) when treated with streptavidin-HRP. The staining appears over the entire glass surface not only where the cells and OCT were. This all-over-staining also appears on clean (without OCT on them) slides that are processed with the OCT slides. I have tried extensive water washes and the results remain the same. No primary antibody is added and the results are the same. Have you seen this before? Do you have any suggestions? Thank you for your help. ~Emily Emily A. Smith CuraGen Corporation 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 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 jtaylor <@t> meriter.com Wed May 19 10:16:22 2004 From: jtaylor <@t> meriter.com (Taylor, Jean) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] p16INK4a Message-ID: <328CBAE62F31C642B422970E879DFADC093CB9@pcwex01.meriter.com> The Pathologists I work for have requested that I purchase p16INK4a. I'd like to get an idea of which company most people order from and whether you use the monoclonal or polyclonal antibody. Thanks, Jean Taylor Immuno Tech GML Madison, WI -----Original Message----- From: Margaret Blount [mailto:mab70@medschl.cam.ac.uk] Sent: Wednesday, May 19, 2004 09:58 To: 'Smith, Emily'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Background Staining Dear Emily, That's a strange one. Have you titrated your secondary antibody sufficiently? what blocking solution do you use? Have the slides inadvertently dried out at any stage of the procedure? These are a few thoughts that spring to mind. The other thing that I would look at would be to try charged slides from another supplier, or try Polysine or APES slides for comparison on indeed another batch of slides. That's my 2 penny worth - maybe someone else will have something more helpful to say and no doubt you have thought of some or all of these ideas yourself. Good luck. Regards Margaret Margaret Blount Chief Technician Clinical Biochemistry University of Cambridge Addenbrooke's Hospital Hills Road Cambridge CB2 2QR -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Smith, Emily Sent: Wednesday, May 19, 2004 3:42 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Background Staining Hello All, In the course of a recent experiment, I saw strong background staining on charged slides with frozen cells in OCT (optimal cutting temperature) when treated with streptavidin-HRP. The staining appears over the entire glass surface not only where the cells and OCT were. This all-over-staining also appears on clean (without OCT on them) slides that are processed with the OCT slides. I have tried extensive water washes and the results remain the same. No primary antibody is added and the results are the same. Have you seen this before? Do you have any suggestions? Thank you for your help. ~Emily Emily A. Smith CuraGen Corporation 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 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Wed May 19 10:18:55 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] H and E Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0635A76@UTHEVS3.mail.uthouston.edu> I personally do not like either Harris or Gill's hematoxylin as I was brought up using hand staining and Ehrlich's hematoxylin made from scratch. There are a lot of individuals using both these solution in Histoland, so I have to assume that they can't all be wrong. There have been a lot of hematoxylins formulated by individuals from several cultures (not just dead white guys), after all hematoxylin was used in South America for dyeing cloth way before anyone thought of using it for staining tissue sections. Recently Placido Domingo, who I think we can all agree is a great tenor, when being interviewed on TV said "I do not understand rap but that is my problem, I do respect it". I think that this is a valuable lesson from a great master. Let us not knock the younger generation just because they do not necessarily have our principles or standards. It is arrogant of us to regard ours as being the only ones possible or desirable. I do believe that it is common sense to foster innovation. I am 65 but I find that I can always learn something from "the new generation", life before 30 is not necessarily wasted and life does not end after 30. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Connie McManus Sent: Wednesday, May 19, 2004 8:49 AM To: 'Gary Gill'; 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Gary Is Gill a relative or yours??? I don't give a hoot who made what when. I like Harris. I don't believe Gill has made any significant improvements over the old dead white guy's hematoxylin. BTW, classical music may have been composed by dead white guys, but I don't hear ANYTHING being composed today that has the depth, complexity and color of those great composers. I've studied piano from since I was 4 yrs old, and I've studied the organ (I've played as a church organist)for almost 20 years. I know music like the back of my hand and I love ALL music (except gangstah stuff). SO DON'T try to tell me about what music is living and worth my while. Dead white Europeans. Yeah, right. I'd like to see someone between 8 and 14 in this generation compose music like Felix Mendelsohnn's or Mozart's. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Tuesday, May 18, 2004 10:04 AM To: 'Connie McManus'; 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems You may have heard that classical music was composed by dead white Europeans. Well, Harris hematoxylin was composed by a dead white American (physician at Jefferson Hospital in 1904). Gill hematoxylin was composed by a live white American. So if you want to liven things up, go with Gill's! Gary Gill (one and the same) PS -- No royalties involved, thanks to bad advice in 1972 from corporate counsel for Johns Hopkins Medical School. -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Tuesday, May 18, 2004 10:41 AM To: 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Wow. What a lot of interesting comments!! I agree with Terry re the agitation. When I watch the stainer do those dips (I can program how many, but NOT the briskness), I wonder if you could even call it agitation. My hand dips are very brisk. Also, I don't bother letting the slides stay in the alcohols for 1 minute or 2, I give the slides about 20 -30 good brisk dips in each solution, then the timed rinses & staining. This has always been far more satisfactory to me than those sllllooooowwwww dips from the machine. As for the kind of hematoxylin, someone suggested I throw out the Harris and do Gills III. I've tried Gill III before and I much prefer the Harris. So it's just a matter of personal preference on that... AND what your pathologist likes *G* Everyone having an nice Tuesday??? *g* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 ----- From StarkusL <@t> ummhc.org Wed May 19 10:34:19 2004 From: StarkusL <@t> ummhc.org (Starkus, Laurie) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] H and E Message-ID: Barry, Very well said. Laurie -----Original Message----- From: Barry R Rittman [mailto:Barry.R.Rittman@uth.tmc.edu] Sent: Wednesday, May 19, 2004 11:19 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] H and E I personally do not like either Harris or Gill's hematoxylin as I was brought up using hand staining and Ehrlich's hematoxylin made from scratch. There are a lot of individuals using both these solution in Histoland, so I have to assume that they can't all be wrong. There have been a lot of hematoxylins formulated by individuals from several cultures (not just dead white guys), after all hematoxylin was used in South America for dyeing cloth way before anyone thought of using it for staining tissue sections. Recently Placido Domingo, who I think we can all agree is a great tenor, when being interviewed on TV said "I do not understand rap but that is my problem, I do respect it". I think that this is a valuable lesson from a great master. Let us not knock the younger generation just because they do not necessarily have our principles or standards. It is arrogant of us to regard ours as being the only ones possible or desirable. I do believe that it is common sense to foster innovation. I am 65 but I find that I can always learn something from "the new generation", life before 30 is not necessarily wasted and life does not end after 30. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Connie McManus Sent: Wednesday, May 19, 2004 8:49 AM To: 'Gary Gill'; 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Gary Is Gill a relative or yours??? I don't give a hoot who made what when. I like Harris. I don't believe Gill has made any significant improvements over the old dead white guy's hematoxylin. BTW, classical music may have been composed by dead white guys, but I don't hear ANYTHING being composed today that has the depth, complexity and color of those great composers. I've studied piano from since I was 4 yrs old, and I've studied the organ (I've played as a church organist)for almost 20 years. I know music like the back of my hand and I love ALL music (except gangstah stuff). SO DON'T try to tell me about what music is living and worth my while. Dead white Europeans. Yeah, right. I'd like to see someone between 8 and 14 in this generation compose music like Felix Mendelsohnn's or Mozart's. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Tuesday, May 18, 2004 10:04 AM To: 'Connie McManus'; 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems You may have heard that classical music was composed by dead white Europeans. Well, Harris hematoxylin was composed by a dead white American (physician at Jefferson Hospital in 1904). Gill hematoxylin was composed by a live white American. So if you want to liven things up, go with Gill's! Gary Gill (one and the same) PS -- No royalties involved, thanks to bad advice in 1972 from corporate counsel for Johns Hopkins Medical School. -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Tuesday, May 18, 2004 10:41 AM To: 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Wow. What a lot of interesting comments!! I agree with Terry re the agitation. When I watch the stainer do those dips (I can program how many, but NOT the briskness), I wonder if you could even call it agitation. My hand dips are very brisk. Also, I don't bother letting the slides stay in the alcohols for 1 minute or 2, I give the slides about 20 -30 good brisk dips in each solution, then the timed rinses & staining. This has always been far more satisfactory to me than those sllllooooowwwww dips from the machine. As for the kind of hematoxylin, someone suggested I throw out the Harris and do Gills III. I've tried Gill III before and I much prefer the Harris. So it's just a matter of personal preference on that... AND what your pathologist likes *G* Everyone having an nice Tuesday??? *g* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 ----- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Wed May 19 09:30:07 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] H&E stain problems Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0635A73@UTHEVS3.mail.uthouston.edu> I personally do not like either Harris or Gill's hematoxylin as I was brought up using hand staining and Ehrlich's hematoxylin made from scratch. There are a lot of individuals using both these solution in Histoland, so I have to assume that they can't all be wrong. There have been a lot of hematoxylins formulated by individuals from several cultures (not just dead white guys), after all hematoxylin was used in South America for dyeing cloth way before anyone thought of using it for staining tissue sections. Recently Placido Domingo, who I think we can all agree is a great tenor, when being interviewed on TV said "I do not understand rap but that is my problem, I do respect it". I think that this is a valuable lesson from a great master. Let us not knock the younger generation just because they do not necessarily have our principles or standards. It is arrogant of us to regard ours as being the only ones possible or desirable. I do believe that it is common sense to foster innovation. I am 65 but I find that I can always learn something from "the new generation", life does not necessarily end after 30. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Connie McManus Sent: Wednesday, May 19, 2004 8:49 AM To: 'Gary Gill'; 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Gary Is Gill a relative or yours??? I don't give a hoot who made what when. I like Harris. I don't believe Gill has made any significant improvements over the old dead white guy's hematoxylin. BTW, classical music may have been composed by dead white guys, but I don't hear ANYTHING being composed today that has the depth, complexity and color of those great composers. I've studied piano from since I was 4 yrs old, and I've studied the organ (I've played as a church organist)for almost 20 years. I know music like the back of my hand and I love ALL music (except gangstah stuff). SO DON'T try to tell me about what music is living and worth my while. Dead white Europeans. Yeah, right. I'd like to see someone between 8 and 14 in this generation compose music like Felix Mendelsohnn's or Mozart's. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Tuesday, May 18, 2004 10:04 AM To: 'Connie McManus'; 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems You may have heard that classical music was composed by dead white Europeans. Well, Harris hematoxylin was composed by a dead white American (physician at Jefferson Hospital in 1904). Gill hematoxylin was composed by a live white American. So if you want to liven things up, go with Gill's! Gary Gill (one and the same) PS -- No royalties involved, thanks to bad advice in 1972 from corporate counsel for Johns Hopkins Medical School. -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Tuesday, May 18, 2004 10:41 AM To: 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Wow. What a lot of interesting comments!! I agree with Terry re the agitation. When I watch the stainer do those dips (I can program how many, but NOT the briskness), I wonder if you could even call it agitation. My hand dips are very brisk. Also, I don't bother letting the slides stay in the alcohols for 1 minute or 2, I give the slides about 20 -30 good brisk dips in each solution, then the timed rinses & staining. This has always been far more satisfactory to me than those sllllooooowwwww dips from the machine. As for the kind of hematoxylin, someone suggested I throw out the Harris and do Gills III. I've tried Gill III before and I much prefer the Harris. So it's just a matter of personal preference on that... AND what your pathologist likes *G* Everyone having an nice Tuesday??? *g* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, May 18, 2004 7:35 AM To: Connie McManus; Petia P Stefanova; Megan Kear; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Connie remarks: "In truth, I prefer my hand stained sections better than when they're stained automatically." When I first saw what I call x-y stainers, I thought that we had in this, something that reproduced hand staining. Not so. What I think is lacking is the brisk agitation necessary to break down the interface between old and new solution. I'm not so sure about the rinsing either. 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 18 May 2004 15:09 To: 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ 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 asmith <@t> mail.barry.edu Wed May 19 10:08:18 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Efficiency of p o and m xylenes in dewaxing Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3B32@exchsrv01.barrynet.barry.edu> I have never tried the pure isomers myself, but three of my professors told me that all 3 or any mixture thereof were equally effective. (They probably just heard it from their professors.) The mixed xylenes (which also contain some ethylbenzene) are a little cheaper than m-xylene and much cheaper than the other pure isomers. 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 Tony Henwood Sent: Tuesday, May 18, 2004 9:09 PM To: Histonet (Histonet@lists.utsouthwestern.edu); 'histonet@pathology.swmed.edu' Subject: [Histonet] Efficiency of p o and m xylenes in dewaxing Hi all, Would anyone have any information on the relative efficiencies of para, meta and ortho xylenes in dewaxing paraffin sections for nucleic acid extraction. Does it matter? 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 ********************************************************************** 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 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 mbryhan <@t> NORTHERNHEALTH.ORG Wed May 19 10:22:10 2004 From: mbryhan <@t> NORTHERNHEALTH.ORG (Mary Bryhan) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] Alternative fixative users (Prefer) Message-ID: Hi, after many years as a Prefer fixative user our pathologists are telling us that we have to go back to formalin. This came about after a letter to the editor appeared in the Journal of Surgical Pathology. Has anyone out there been approached by their pathologists to switch back? Mary Bryhan From Barry.R.Rittman <@t> uth.tmc.edu Wed May 19 09:52:38 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] help Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0635A74@UTHEVS3.mail.uthouston.edu> Blanche This may be somewhat of a mute point as many of the fluorescent markers fade over time, sometimes over very short times. This is especially true when exposed to viewing illumination for some time. It also depends on the storage conditions, are these slides frozen, paraffin, what mountant if any are they in and what conditions of storage? It would be best if a record is required to photograph the image on which the diagnosis is made and store this image. I realize that for many labs this may not be a possibility. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Williams, Blanche Sent: Wednesday, May 19, 2004 9:23 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] help Can someone answer a question regarding slide retention for me? How long to labs keep their immunofluorescence slides? -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Wednesday, May 19, 2004 10:16 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 6, Issue 28 From GAshton <@t> PICR.man.ac.uk Wed May 19 10:32:59 2004 From: GAshton <@t> PICR.man.ac.uk (Garry Ashton) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] HIF 1 alpha Message-ID: Dear all, I'm having problems with an antibody we are using. The antibody is HIF 1alpha. We have had some good staining, and then some poor results. It was mentioned that the age of the sections could have something to do with it. Has any experienced this before, or know of any other reason. The antibody has been stored as instructed and is well within date. Many thanks for any help in advance. Garry PICR UK -------------------------------------------------------- This email is confidential and intended solely for the use of the person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Paterson Institute for Cancer Research or the Christie Hospital NHS Trust. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From pengbw <@t> sjtu.edu.cn Wed May 19 10:29:01 2004 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] about immunofluonence staining Message-ID: <20040519152901.E268A111A3DF@sjtu.edu.cn> Hi,all histoneter I would like to know if I could do immunofluonence staining like the fellowing, I used a system include a first antiboy, a biotinlate second antibody and a streptavidin conjugated FITC insteed of using a FITC conjugated antibody to antigen. And you know, I\'m doing so just want to save my boss\'s money. But I have no confident on my system even though it works. Baowei Peng Shanghai Jiaotong University Shanghai, 200030 China From Terry.Marshall <@t> rothgen.nhs.uk Wed May 19 10:43:57 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:59 2005 Subject: [Histonet] H and E Message-ID: Ah Barry - life has become instant everything. Why look longingly at immature H on a windowsill for months when in a flash of periodate, it can be yours, ready, now? My first experience with Ehrlich's, apart from school, was in my final professional exam in Liverpool, where everything was stained with it. I found it so difficult with all the mucin and ground substance (still mucin I know) staining blue. Particularly irksome when I like my mucin green:-) I have always found Gill's to be much as Harris' but more reliable, at least, if you make it yourself. 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: Barry R Rittman [mailto:Barry.R.Rittman@uth.tmc.edu] Sent: 19 May 2004 16:19 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] H and E I personally do not like either Harris or Gill's hematoxylin as I was brought up using hand staining and Ehrlich's hematoxylin made from scratch. There are a lot of individuals using both these solution in Histoland, so I have to assume that they can't all be wrong. There have been a lot of hematoxylins formulated by individuals from several cultures (not just dead white guys), after all hematoxylin was used in South America for dyeing cloth way before anyone thought of using it for staining tissue sections. Recently Placido Domingo, who I think we can all agree is a great tenor, when being interviewed on TV said "I do not understand rap but that is my problem, I do respect it". I think that this is a valuable lesson from a great master. Let us not knock the younger generation just because they do not necessarily have our principles or standards. It is arrogant of us to regard ours as being the only ones possible or desirable. I do believe that it is common sense to foster innovation. I am 65 but I find that I can always learn something from "the new generation", life before 30 is not necessarily wasted and life does not end after 30. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Connie McManus Sent: Wednesday, May 19, 2004 8:49 AM To: 'Gary Gill'; 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Gary Is Gill a relative or yours??? I don't give a hoot who made what when. I like Harris. I don't believe Gill has made any significant improvements over the old dead white guy's hematoxylin. BTW, classical music may have been composed by dead white guys, but I don't hear ANYTHING being composed today that has the depth, complexity and color of those great composers. I've studied piano from since I was 4 yrs old, and I've studied the organ (I've played as a church organist)for almost 20 years. I know music like the back of my hand and I love ALL music (except gangstah stuff). SO DON'T try to tell me about what music is living and worth my while. Dead white Europeans. Yeah, right. I'd like to see someone between 8 and 14 in this generation compose music like Felix Mendelsohnn's or Mozart's. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Tuesday, May 18, 2004 10:04 AM To: 'Connie McManus'; 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems You may have heard that classical music was composed by dead white Europeans. Well, Harris hematoxylin was composed by a dead white American (physician at Jefferson Hospital in 1904). Gill hematoxylin was composed by a live white American. So if you want to liven things up, go with Gill's! Gary Gill (one and the same) PS -- No royalties involved, thanks to bad advice in 1972 from corporate counsel for Johns Hopkins Medical School. -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Tuesday, May 18, 2004 10:41 AM To: 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Wow. What a lot of interesting comments!! I agree with Terry re the agitation. When I watch the stainer do those dips (I can program how many, but NOT the briskness), I wonder if you could even call it agitation. My hand dips are very brisk. Also, I don't bother letting the slides stay in the alcohols for 1 minute or 2, I give the slides about 20 -30 good brisk dips in each solution, then the timed rinses & staining. This has always been far more satisfactory to me than those sllllooooowwwww dips from the machine. As for the kind of hematoxylin, someone suggested I throw out the Harris and do Gills III. I've tried Gill III before and I much prefer the Harris. So it's just a matter of personal preference on that... AND what your pathologist likes *G* Everyone having an nice Tuesday??? *g* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 ----- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> colobio.com Wed May 19 11:02:08 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] help In-Reply-To: <85151E3A0A49DF498F39F19EE56FEA8D0317CECF@lsexch2.lsmaster.lifespan.org> Message-ID: We always photographed them asap and then got rid of them, they do fade and take up space in the fridge. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Williams, Blanche Sent: Wednesday, May 19, 2004 8:23 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] help Can someone answer a question regarding slide retention for me? How long to labs keep their immunofluorescence slides? -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Wednesday, May 19, 2004 10:16 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 6, Issue 28 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. CJD DECONTAMINATION PROCEDURE (srishan@mail.holyname.org) 2. RE: H&E stain problems (Gary Gill) 3. RE: CJD DECONTAMINATION PROCEDURE (Bartlett, Jeanine) 4. RE: CJD DECONTAMINATION PROCEDURE (Kari Bradshaw) 5. research charges (Diane M Nelson) 6. RE: CJD DECONTAMINATION PROCEDURE (Monson, Frederick ) 7. Presto Fix - ? (Richard Cartun) 8. Re: Aussies and Kiwis standing up at microtomes (P. Emry) 9. Congo Red (Karen Wittler) 10. Re: MIcrotome alignment (Ford Royer) 11. RE: research charges (Mass Histology Service) 12. Efficiency of p o and m xylenes in dewaxing (Tony Henwood) 13. Efficiency of p o and m xylenes in dewaxing (Tony Henwood) 14. HTL tissue size minimum requriements (Tara Mciver) 15. unsubscribe (Weems, Joyce) 16. (no subject) (yan gao) 17. (no subject) (yan gao) 18. (no subject) (yan gao) 19. RE: HTL tissue size minimum requriements (Galbraith, Joe) 20. (no subject) (phyllis) 21. RE: H&E stain problems (Connie McManus) 22. RE: H&E stain problems (Marshall Terry Dr, Consultant Histopathologist) 23. RE: CJD DECONTAMINATION PROCEDURE (Connie McManus) ---------------------------------------------------------------------- Message: 1 Date: Tue, 18 May 2004 12:58:33 -0400 From: srishan@mail.holyname.org Subject: [Histonet] CJD DECONTAMINATION PROCEDURE To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Hello everyone, This question might have been posted before, but, Histology Department is expecting a brain biopsy from a possible CJD patient today. The specimen will be coming down in formalin from the O.R. It will be kept in formalin for 2-7 days and will be transferred to formic acid for the required time. Then the specimen will be transferred to formalin before processing for another 2 days. Has anyone dealt with this procedure before. These are the questions asked by the hstology department. How do you decontaminate the processor, microtome, stainer and coverslipper? What kind of protective gear apart from standard precaution has to be used? How about the shaving of the tissues? How do you dispose the formalin from the original formalin container where the specimen was sent in and how do you dispose the reagents from the processor and the stainer? Can this biopsy be processed, once treated with formic acid, along with the other specimens or should the biopsy be processed with the other specimens? Need Help!! Thank you in advance Mala Srishan Histology Supervisor Holy Name Hospital Teaneck, NJ 07666 ------------------------------ Message: 2 Date: Tue, 18 May 2004 12:03:43 -0500 From: Gary Gill Subject: RE: [Histonet] H&E stain problems To: 'Connie McManus' , "'Marshall Terry Dr,Consultant Histopathologist'" , 'Petia P Stefanova' , 'Megan Kear' , histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain You may have heard that classical music was composed by dead white Europeans. Well, Harris hematoxylin was composed by a dead white American (physician at Jefferson Hospital in 1904). Gill hematoxylin was composed by a live white American. So if you want to liven things up, go with Gill's! Gary Gill (one and the same) PS -- No royalties involved, thanks to bad advice in 1972 from corporate counsel for Johns Hopkins Medical School. -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Tuesday, May 18, 2004 10:41 AM To: 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Wow. What a lot of interesting comments!! I agree with Terry re the agitation. When I watch the stainer do those dips (I can program how many, but NOT the briskness), I wonder if you could even call it agitation. My hand dips are very brisk. Also, I don't bother letting the slides stay in the alcohols for 1 minute or 2, I give the slides about 20 -30 good brisk dips in each solution, then the timed rinses & staining. This has always been far more satisfactory to me than those sllllooooowwwww dips from the machine. As for the kind of hematoxylin, someone suggested I throw out the Harris and do Gills III. I've tried Gill III before and I much prefer the Harris. So it's just a matter of personal preference on that... AND what your pathologist likes *G* Everyone having an nice Tuesday??? *g* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, May 18, 2004 7:35 AM To: Connie McManus; Petia P Stefanova; Megan Kear; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Connie remarks: "In truth, I prefer my hand stained sections better than when they're stained automatically." When I first saw what I call x-y stainers, I thought that we had in this, something that reproduced hand staining. Not so. What I think is lacking is the brisk agitation necessary to break down the interface between old and new solution. I'm not so sure about the rinsing either. 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 18 May 2004 15:09 To: 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ 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 ------------------------------ Message: 3 Date: Tue, 18 May 2004 13:58:07 -0400 From: "Bartlett, Jeanine" Subject: RE: [Histonet] CJD DECONTAMINATION PROCEDURE To: , Message-ID: Content-Type: text/plain; charset="us-ascii" The WHO booklet was too large to send as an attachment so it couldn't be delivered. I will try and send you what I think is the most relevant chapter. Perhaps you could visit the WHO site and get more information if needed. In the meantime, the Surveillance Center link below should have enough information. Jeanine - Mala: There is a lot of conflicting information out there but here is information from 2 very credible sources for you to review. One is the National Surveillance Center at case Western and the other is the WHO. Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 http://www.cjdsurveillance.com and the WHO booklet is sent as an attachment. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of srishan@mail.holyname.org Sent: Tuesday, May 18, 2004 12:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CJD DECONTAMINATION PROCEDURE Hello everyone, This question might have been posted before, but, Histology Department is expecting a brain biopsy from a possible CJD patient today. The specimen will be coming down in formalin from the O.R. It will be kept in formalin for 2-7 days and will be transferred to formic acid for the required time. Then the specimen will be transferred to formalin before processing for another 2 days. Has anyone dealt with this procedure before. These are the questions asked by the hstology department. How do you decontaminate the processor, microtome, stainer and coverslipper? What kind of protective gear apart from standard precaution has to be used? How about the shaving of the tissues? How do you dispose the formalin from the original formalin container where the specimen was sent in and how do you dispose the reagents from the processor and the stainer? Can this biopsy be processed, once treated with formic acid, along with the other specimens or should the biopsy be processed with the other specimens? Need Help!! Thank you in advance Mala Srishan Histology Supervisor Holy Name Hospital Teaneck, NJ 07666 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Tue, 18 May 2004 10:55:17 -0700 From: "Kari Bradshaw" Subject: RE: [Histonet] CJD DECONTAMINATION PROCEDURE To: , Message-ID: Content-Type: text/plain; charset="US-ASCII" These people have all the answers: National Prion Pathology Surveillance Center Case Western Reserve University 2085 Adelbert Road Cleveland,Ohio 44106 216-368-0587 www.cjdsurveillance.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of srishan@mail.holyname.org Sent: Tuesday, May 18, 2004 9:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CJD DECONTAMINATION PROCEDURE Hello everyone, This question might have been posted before, but, Histology Department is expecting a brain biopsy from a possible CJD patient today. The specimen will be coming down in formalin from the O.R. It will be kept in formalin for 2-7 days and will be transferred to formic acid for the required time. Then the specimen will be transferred to formalin before processing for another 2 days. Has anyone dealt with this procedure before. These are the questions asked by the hstology department. How do you decontaminate the processor, microtome, stainer and coverslipper? What kind of protective gear apart from standard precaution has to be used? How about the shaving of the tissues? How do you dispose the formalin from the original formalin container where the specimen was sent in and how do you dispose the reagents from the processor and the stainer? Can this biopsy be processed, once treated with formic acid, along with the other specimens or should the biopsy be processed with the other specimens? Need Help!! Thank you in advance Mala Srishan Histology Supervisor Holy Name Hospital Teaneck, NJ 07666 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Tue, 18 May 2004 13:03:49 -0500 From: Diane M Nelson Subject: [Histonet] research charges To: histonet@lists.utsouthwestern.edu Message-ID: <6.1.0.6.2.20040518124918.01b5caa0@dmnelson.mail.iastate.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Hi Histonetters! Our veterinary lab is in the process of up dating our research charges. I would like to know what other labs/hospitals charge for H & E bone slides and H & E eye slides. Currently we are charging, Bone: $19.00 for decalcifying in 25% formic acid, process, embed, sectioning and H & E stain. We are losing money on our larger bone projects. Our charges for processing, embedding sectioning and H & E stain for eye slides is $9.00. It also depends on the project rather or not we make or lose money. The researchers do their own grossing of tissue. Any help would be greatly appreciated! Thanks in advance. Diane Gerjets Iowa State University ------------------------------ Message: 6 Date: Tue, 18 May 2004 14:04:45 -0400 From: "Monson, Frederick " Subject: RE: [Histonet] CJD DECONTAMINATION PROCEDURE To: Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" http://www.nursingceu.com/NCEU/courses/prions/ http://www.mad-cow.org/tonsil_test2.html#Biosafety A start. Cheers Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone/FAX: 610-738-0437 eMail: fmonson@wcupa.edu CASI Page and Scheduling http://darwin.wcupa.edu/CASI/ -----Original Message----- From: srishan@mail.holyname.org [mailto:srishan@mail.holyname.org] Sent: Tuesday, May 18, 2004 12:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CJD DECONTAMINATION PROCEDURE Hello everyone, This question might have been posted before, but, Histology Department is expecting a brain biopsy from a possible CJD patient today. The specimen will be coming down in formalin from the O.R. It will be kept in formalin for 2-7 days and will be transferred to formic acid for the required time. Then the specimen will be transferred to formalin before processing for another 2 days. Has anyone dealt with this procedure before. These are the questions asked by the hstology department. How do you decontaminate the processor, microtome, stainer and coverslipper? What kind of protective gear apart from standard precaution has to be used? How about the shaving of the tissues? How do you dispose the formalin from the original formalin container where the specimen was sent in and how do you dispose the reagents from the processor and the stainer? Can this biopsy be processed, once treated with formic acid, along with the other specimens or should the biopsy be processed with the other specimens? Need Help!! Thank you in advance Mala Srishan Histology Supervisor Holy Name Hospital Teaneck, NJ 07666 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Tue, 18 May 2004 14:32:48 -0400 From: "Richard Cartun" Subject: [Histonet] Presto Fix - ? To: Message-ID: Content-Type: text/plain; charset=US-ASCII Anyone using "Presto Fix" for bone marrow core biopsies? If so, what does it do to immunoreactivity? Thank you. Richard Cartun ------------------------------ Message: 8 Date: Tue, 18 May 2004 11:34:18 -0700 (PDT) From: "P. Emry" Subject: Re: [Histonet] Aussies and Kiwis standing up at microtomes To: Andrew Kennedy Cc: Histonet Message-ID: Content-Type: TEXT/PLAIN; charset=US-ASCII Ahhhhh working with a beer at my elbow. Sounds very sane to me. I think we got one of your country women..a Wealthal..up for a phd. She denies dwarf tossing, but I have heard things to the contrary. I have two square carts with drawers in used in the dental school. They have wheels on them. I put the microtome on one and the waterbath on the other. They sit side-by side with a space between. They are just high enough for me to use a standard height chair with wheels and move back and forth between them. It is the best arrangement I have found. I can get up over the water bath to get a good look at what is going on. I really like it. Not hard on the back like benches. Would suggest it for those having problems with benches and sitting still in one possition. If I can figure out how to put an ice bucket for the beer between, we's really have something. Trisha On Mon, 17 May 2004, Andrew Kennedy wrote: > Hi Tim and Fred (and the rest of Histonetters too!!) > > > > In my experience, we Aussies don't like to stand up at the microtome if we > can help it. Standing and cutting sections makes no sense at all - we might > spill our beer and not be able to see the television that sits on the bench > next to us so we can watch Aussie Rules (I prefer Rugby League, by the way) > > :-) > > But seriously folks, you would have to have high benches and even then you > would probably hunch over the microtome - I remember seeing a histotech > cutting standing up years ago when I was a lab assistant. She turned the > microtome sideways, stood at the bench (which was desk height) and stooped > over the microtome. That can't be good for your back! It was the most > uncomfortable looking cutting position I've ever seen. > > > > Overlapping with the safety thread that is currently quite "hot", we really > owe it to ourselves and to those we work with to be safe all the time! > > > > In Australia, Occupational Health and Safety is an important issue that > affects everything we do in the workplace. However, any safety procedure > that has been implemented in our labs is a result of consultation with the > parties at stake. Safe Operating Procedures are designed by the people who > will use them, not by some beaurocrat sitting in an office telling others > what to do! Take control of your safety and use common sense at the same > time! Makes sense to me!! > > > > Andrew Kennedy > > Senior Science Officer > > Anatomical Pathology > > Concord Repatriation General Hospital > > Hospital Road > Concord NSW Australia 2139 > > > > ph: +612 9767 6115 > > Fax +612 9767 8427 > > > > "oderint dum metuant" > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 9 Date: Tue, 18 May 2004 16:29:47 -0400 From: Karen Wittler Subject: [Histonet] Congo Red To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII We stain our Congo Reds on an automated stainer now. Before that we stained with hematoxylin first because rinsing and differentiation was required and we used an alkaline Congo Red. After staining with the Congo Red in a saturated sodium chloride- alcohol solution,we did not rinse,but rapidly dehydrated, cleared and mounted For our H&Es we use Modified Harris Hematoxylin and Eosin-Y w/ Phloxine both manufactured by Richard Allen. Karen Wittler, HT(ASCP) Johns Hopkins Hospital Baltimore, MD ------------------------------ Message: 10 Date: Tue, 18 May 2004 16:16:32 -0500 From: "Ford Royer" Subject: Re: [Histonet] MIcrotome alignment To: KHays@mbhs.org, histonet@lists.utsouthwestern.edu Cc: Marcia Welch Message-ID: <40AA7D30.10601@bitstream.net> Content-Type: text/plain; charset=us-ascii; format=flowed On their web site catalog, they show a price of $775.00. Contact Marcia Welch at: 800-383-7799 or Email: for a customer price quote. Ford M. Royer, MT(ASCP) Analytical Instruments, llc 1200 Mendelssohn Ave. N., Ste. 50 Minneapolis, MN 55427 (800) 565-1895, ext. 17 URL: KHays@mbhs.org wrote: > > Kathy Tedford-Hays HT (ASCP) > Technical Specialist, Histology Dept > (601)-968-3070 ext 7398 > Baptist Medical Center > 1225 North State Street > Jackson, MS 39202 > > how much is it , do you know? > > > > "Ford Royer" > Sent by: histonet-bounces@lists.utsouthwestern.edu > > 05/18/2004 09:00 AM > > > To: histonet@lists.utsouthwestern.edu > cc: > Fax to: > Subject: Re: [Histonet] MIcrotome alignment > > > > > Newcomer Supply, Madison, WI sells a microtome aligner. Product Code > 6255A > Web URL > Check it out. > > ~ Ford > Ford M. Royer, MT(ASCP) > Analytical Instruments, llc > Minneapolis, MN 55427 > (800) 565-1895, Ext. 17 > > > Roberts, Jon wrote: > > >I have recently purchased a Leica RM2255 with the new precise > specimen orientation system that adjust to an exact zero position or > x-y axis standard. But I also have old leicas without this system. > Is there any quick and easy way to align my old leicas to the new > one???? Just wondering? I though I've heard of such a device, but > wonder if anyone else has heard of it or actually used it? > > > > > >CONFIDENTIALITY NOTICE: This email message and any accompanying data > are confidential, and intended only for the named recipient(s). If > you are not the intended recipient(s), you are hereby notified that > the dissemination, distribution, and or copying of this message is > strictly prohibited. If you receive this message in error, or are not > the named recipient(s), please notify the sender at the email address > above, delete this email from your computer, and destroy any copies in > any form 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 > This internet email has been certified virus free by BHS Anti-Virus > system. > > ------------------------------ Message: 11 Date: Tue, 18 May 2004 19:59:45 -0400 From: "Mass Histology Service" Subject: RE: [Histonet] research charges To: "Diane M Nelson" , Message-ID: Content-Type: text/plain; charset="us-ascii" Diane, We charge one price across the board, $5.50 if the tissue is received in a cassette and $2.50 if it needs decalcification. Our philosophy is that we don't charge less for simple specimens, so why charge more for occasional complicated ones? Of course if it's a large project consisting of many whole eyes or femur heads, I'd probably double our current prices to cover for the additional labor and material. Jim ____________________________ James E. Staruk, HT(ASCP) Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Diane M Nelson Sent: Tuesday, May 18, 2004 2:04 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] research charges Hi Histonetters! Our veterinary lab is in the process of up dating our research charges. I would like to know what other labs/hospitals charge for H & E bone slides and H & E eye slides. Currently we are charging, Bone: $19.00 for decalcifying in 25% formic acid, process, embed, sectioning and H & E stain. We are losing money on our larger bone projects. Our charges for processing, embedding sectioning and H & E stain for eye slides is $9.00. It also depends on the project rather or not we make or lose money. The researchers do their own grossing of tissue. Any help would be greatly appreciated! Thanks in advance. Diane Gerjets Iowa State University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Wed, 19 May 2004 11:08:30 +1000 From: Tony Henwood Subject: [Histonet] Efficiency of p o and m xylenes in dewaxing To: "Histonet (Histonet@lists.utsouthwestern.edu)" , "'histonet@pathology.swmed.edu'" Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E1C8@simba.kids> Content-Type: text/plain Hi all, Would anyone have any information on the relative efficiencies of para, meta and ortho xylenes in dewaxing paraffin sections for nucleic acid extraction. Does it matter? 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 ********************************************************************** 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. ********************************************************************** ------------------------------ Message: 13 Date: Wed, 19 May 2004 11:08:30 +1000 From: Tony Henwood Subject: [Histonet] Efficiency of p o and m xylenes in dewaxing To: "Histonet (Histonet@lists.utsouthwestern.edu)" , "'histonet@pathology.swmed.edu'" Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E1C8@simba.kids> Content-Type: text/plain Hi all, Would anyone have any information on the relative efficiencies of para, meta and ortho xylenes in dewaxing paraffin sections for nucleic acid extraction. Does it matter? 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 ********************************************************************** 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. ********************************************************************** ------------------------------ Message: 14 Date: Wed, 19 May 2004 05:50:52 -0400 From: "Tara Mciver" Subject: [Histonet] HTL tissue size minimum requriements To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" I attended a readiness workshop in NC and thought I understood the instructor to say that the tissue sizes should be at least the size indicated, so actually the tissue size could be larger, and that is what he suggested we do. Can anybody verify that I understood him correctly? Also, does the tissue have to be a perfect square, or can I just be sure it covers the square representation provided with the test? Thanks for any help! ------------------------------ Message: 15 Date: Wed, 19 May 2004 07:11:09 -0400 From: "Weems, Joyce" Subject: [Histonet] unsubscribe To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" 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. ------------------------------ Message: 16 Date: Wed, 19 May 2004 09:08:42 -0400 From: "yan gao" Subject: [Histonet] (no subject) To: histonet@lists.utsouthwestern.edu Cc: gliuygao@hotmail.com Message-ID: Content-Type: text/plain Hi, Everyone. I am looking for CD34 antibody stain rat tissues. Anyone knows where I can get it? Thanks. _________________________________________________________________ [1]Express yourself with the new version of MSN Messenger! Download today - it's FREE! References 1. http://g.msn.com/8HMBENUS/2752??PS=47575 ------------------------------ Message: 17 Date: Wed, 19 May 2004 09:09:28 -0400 From: "yan gao" Subject: [Histonet] (no subject) To: histonet@lists.utsouthwestern.edu Cc: gliuygao@hotmail.com Message-ID: Content-Type: text/plain Hi, Everyone. I am looking for CD34 antibody stain rat tissues. Anyone knows where I can get it? Thanks. Yan Gao Doctoral of Science Novartis _________________________________________________________________ [1]Express yourself with the new version of MSN Messenger! Download today - it's FREE! References 1. http://g.msn.com/8HMBENUS/2752??PS=47575 ------------------------------ Message: 18 Date: Wed, 19 May 2004 09:05:25 -0400 From: "yan gao" Subject: [Histonet] (no subject) To: histonet@list.utsouthwestern.edu, histonet@pathology.swmed.edu Cc: gliuygao@hotmail.com Message-ID: Content-Type: text/plain Hi, Everyone. I am looking for CD34 antibody stain rat tissues. Anyone knows where I can get it? Thanks. Yan Gao NOVARTIS _________________________________________________________________ [1]Learn to simplify your finances and your life in Streamline Your Life from MSN Money. References 1. http://g.msn.com/8HMBENUS/2743??PS=47575 ------------------------------ Message: 19 Date: Wed, 19 May 2004 08:10:52 -0500 From: "Galbraith, Joe" Subject: RE: [Histonet] HTL tissue size minimum requriements To: "Tara Mciver" , Message-ID: <5D03ED7B9391D4119D9B0008C76B7B2403005B1F@uihc-mail1.uihc.uiowa.edu> Content-Type: text/plain; charset="iso-8859-1" The sizes are minimums as you understand. You will be held strictly accountable for ensuring that the tissue you submit in your block must exceed the dimensions listed and that the tissue on the slide must match the dimensions in the block (without excessive spreading from overfloating on a warm water bath). Remember to allow for shrinkage during processing when you are acquiring your tissue when fixing in formalin and using standard processing. To get 2.0 cm of duodenum into the block you may have to cut 2.5 cm from the source tissue depending on your processing schedule. But 1.9 cm will not due. Also, if they specify 2.0 cm of duodenum with epitelium along one side, they mean complete uninterupted epithelium in perfect condition, perfectly fixed, perfectly processed, perfectly cut, and perfectly stained. Any flaws will cost you points. I don't mean to be negative just to remind everyone preparing for the practical that you must pay attention to all the details. Read the directions very carefully and follow every picky detail that they specify. A significant part of the exam is the applicant's ability to follow written instructions accurately. (Meaning, if they say a square, I'd recommend a square.) Pay attention to the labeling of your slides. Be sure your labels are clear and readable (typed), labeled exactly as instructed, and that your label material is in good condition (stays on the slide, no curls or peeling, etc.) Hope this helps, good luck. Joe Galbraith -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Tara Mciver Sent: Wednesday, May 19, 2004 4:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HTL tissue size minimum requriements I attended a readiness workshop in NC and thought I understood the instructor to say that the tissue sizes should be at least the size indicated, so actually the tissue size could be larger, and that is what he suggested we do. Can anybody verify that I understood him correctly? Also, does the tissue have to be a perfect square, or can I just be sure it covers the square representation provided with the test? Thanks for any help! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 20 Date: Wed, 19 May 2004 09:46:55 -0400 From: phyllis Subject: [Histonet] (no subject) To: histonet@lists.utsouthwestern.edu Message-ID: <5.1.0.14.0.20040519094604.03b9c2e0@pop.cwru.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Why are you handling this processing? The CJD Surveillance has been set up by CDC to do all this work for you and also provide a diagnosis. If your pathologists choose to do this themselves, be aware of the protocols which can be found at www.cdc.gov and also the WHO website. The biopsy should be handled in a level 2 facility. You need to wear gloves and face shield. All processing and staining is to been done by hand in individual containers which will be incinirated on completion, otherwise you will have deal with the demanding work of decontaminating the processor. All waste is incinerated. You will find all this info at CDC website. Also refer to www.cjdsurveillance.com if you decide to let the CJD Survellance do this for you. Free of charge. Phyllis Scalzo, HT(ASCP) Case Western Reserve University 2085 Adelbert Rd. Cleveland, Ohio 44106 Ph: 216-368-0822 Fax: 216-368-2546 ------------------------------ Message: 21 Date: Wed, 19 May 2004 07:48:52 -0600 From: "Connie McManus" Subject: RE: [Histonet] H&E stain problems To: "'Gary Gill'" , "'Marshall Terry Dr, Consultant Histopathologist'" , "'Petia P Stefanova'" , "'Megan Kear'" , Cc: histonet@lists.utsouthwestern.edu Message-ID: <000a01c43da8$08034a90$4a737b81@Cygnus> Content-Type: text/plain; charset="us-ascii" Gary Is Gill a relative or yours??? I don't give a hoot who made what when. I like Harris. I don't believe Gill has made any significant improvements over the old dead white guy's hematoxylin. BTW, classical music may have been composed by dead white guys, but I don't hear ANYTHING being composed today that has the depth, complexity and color of those great composers. I've studied piano from since I was 4 yrs old, and I've studied the organ (I've played as a church organist)for almost 20 years. I know music like the back of my hand and I love ALL music (except gangstah stuff). SO DON'T try to tell me about what music is living and worth my while. Dead white Europeans. Yeah, right. I'd like to see someone between 8 and 14 in this generation compose music like Felix Mendelsohnn's or Mozart's. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Tuesday, May 18, 2004 10:04 AM To: 'Connie McManus'; 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems You may have heard that classical music was composed by dead white Europeans. Well, Harris hematoxylin was composed by a dead white American (physician at Jefferson Hospital in 1904). Gill hematoxylin was composed by a live white American. So if you want to liven things up, go with Gill's! Gary Gill (one and the same) PS -- No royalties involved, thanks to bad advice in 1972 from corporate counsel for Johns Hopkins Medical School. -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Tuesday, May 18, 2004 10:41 AM To: 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Wow. What a lot of interesting comments!! I agree with Terry re the agitation. When I watch the stainer do those dips (I can program how many, but NOT the briskness), I wonder if you could even call it agitation. My hand dips are very brisk. Also, I don't bother letting the slides stay in the alcohols for 1 minute or 2, I give the slides about 20 -30 good brisk dips in each solution, then the timed rinses & staining. This has always been far more satisfactory to me than those sllllooooowwwww dips from the machine. As for the kind of hematoxylin, someone suggested I throw out the Harris and do Gills III. I've tried Gill III before and I much prefer the Harris. So it's just a matter of personal preference on that... AND what your pathologist likes *G* Everyone having an nice Tuesday??? *g* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, May 18, 2004 7:35 AM To: Connie McManus; Petia P Stefanova; Megan Kear; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Connie remarks: "In truth, I prefer my hand stained sections better than when they're stained automatically." When I first saw what I call x-y stainers, I thought that we had in this, something that reproduced hand staining. Not so. What I think is lacking is the brisk agitation necessary to break down the interface between old and new solution. I'm not so sure about the rinsing either. 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 18 May 2004 15:09 To: 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ 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 ------------------------------ Message: 22 Date: Wed, 19 May 2004 14:56:37 +0100 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] H&E stain problems To: "Connie McManus" , "Gary Gill" , "Petia P Stefanova" , "Megan Kear" , Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" Close relative:-) 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 19 May 2004 14:49 To: 'Gary Gill'; Marshall Terry Dr, Consultant Histopathologist; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Gary Is Gill a relative or yours??? I don't give a hoot who made what when. I like Harris. I don't believe Gill has made any significant improvements over the old dead white guy's hematoxylin. BTW, classical music may have been composed by dead white guys, but I don't hear ANYTHING being composed today that has the depth, complexity and color of those great composers. I've studied piano from since I was 4 yrs old, and I've studied the organ (I've played as a church organist)for almost 20 years. I know music like the back of my hand and I love ALL music (except gangstah stuff). SO DON'T try to tell me about what music is living and worth my while. Dead white Europeans. Yeah, right. I'd like to see someone between 8 and 14 in this generation compose music like Felix Mendelsohnn's or Mozart's. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Tuesday, May 18, 2004 10:04 AM To: 'Connie McManus'; 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems You may have heard that classical music was composed by dead white Europeans. Well, Harris hematoxylin was composed by a dead white American (physician at Jefferson Hospital in 1904). Gill hematoxylin was composed by a live white American. So if you want to liven things up, go with Gill's! Gary Gill (one and the same) PS -- No royalties involved, thanks to bad advice in 1972 from corporate counsel for Johns Hopkins Medical School. -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Tuesday, May 18, 2004 10:41 AM To: 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Wow. What a lot of interesting comments!! I agree with Terry re the agitation. When I watch the stainer do those dips (I can program how many, but NOT the briskness), I wonder if you could even call it agitation. My hand dips are very brisk. Also, I don't bother letting the slides stay in the alcohols for 1 minute or 2, I give the slides about 20 -30 good brisk dips in each solution, then the timed rinses & staining. This has always been far more satisfactory to me than those sllllooooowwwww dips from the machine. As for the kind of hematoxylin, someone suggested I throw out the Harris and do Gills III. I've tried Gill III before and I much prefer the Harris. So it's just a matter of personal preference on that... AND what your pathologist likes *G* Everyone having an nice Tuesday??? *g* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, May 18, 2004 7:35 AM To: Connie McManus; Petia P Stefanova; Megan Kear; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Connie remarks: "In truth, I prefer my hand stained sections better than when they're stained automatically." When I first saw what I call x-y stainers, I thought that we had in this, something that reproduced hand staining. Not so. What I think is lacking is the brisk agitation necessary to break down the interface between old and new solution. I'm not so sure about the rinsing either. 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 18 May 2004 15:09 To: 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ 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 ------------------------------ Message: 23 Date: Wed, 19 May 2004 08:03:16 -0600 From: "Connie McManus" Subject: RE: [Histonet] CJD DECONTAMINATION PROCEDURE To: , Message-ID: <000c01c43daa$087de6e0$4a737b81@Cygnus> Content-Type: text/plain; charset="us-ascii" I don't deal with CJD, but I do a lot of CWD (Chronic Wasting Disease) and Scrapie, which are prion diseases in dear/elk and sheep (respectively). CWD and Scrapie had been associated with humans for hundreds of years with no known transmission of the disease from animal to human. HOWEVER, since prions are not neutralized by fixation of any kind, not killed by autoclaving, this makes them a biohazard in processed tissues. If I worked with CJD, I would ... 1 set up a place that is completely separate from the rest of the lab. Barrier off a section of your lab, use another room (preferable) 2 use disposable bunny suits, shoe covers, face masks, eye wear and DOUBLE GLOVE 3 put stickie mats on the floor completely surrounding the cutting area, 4 put ALL paraffin shavings in a biohaz bag, 5 use paper towels to clean off the microtome and throw those in the biohaz bag 6. incinerate the bunny suit, goggles, gloves, face maskes, etc with the paraffin shavings. 7. Ask for a really big pay raise. Claim it's hazard pay. Have fun. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of srishan@mail.holyname.org Sent: Tuesday, May 18, 2004 9:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CJD DECONTAMINATION PROCEDURE Hello everyone, This question might have been posted before, but, Histology Department is expecting a brain biopsy from a possible CJD patient today. The specimen will be coming down in formalin from the O.R. It will be kept in formalin for 2-7 days and will be transferred to formic acid for the required time. Then the specimen will be transferred to formalin before processing for another 2 days. Has anyone dealt with this procedure before. These are the questions asked by the hstology department. How do you decontaminate the processor, microtome, stainer and coverslipper? What kind of protective gear apart from standard precaution has to be used? How about the shaving of the tissues? How do you dispose the formalin from the original formalin container where the specimen was sent in and how do you dispose the reagents from the processor and the stainer? Can this biopsy be processed, once treated with formic acid, along with the other specimens or should the biopsy be processed with the other specimens? Need Help!! Thank you in advance Mala Srishan Histology Supervisor Holy Name Hospital Teaneck, NJ 07666 _______________________________________________ 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 End of Histonet Digest, Vol 6, Issue 28 *************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From laurie.colbert <@t> huntingtonhospital.com Wed May 19 11:10:44 2004 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Alternative fixative users (Prefer) Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5BF0C@EXCHANGE1.huntingtonhospital.com> What did the letter in the Journal say? Laurie Colbert -----Original Message----- From: Mary Bryhan [mailto:mbryhan@NORTHERNHEALTH.ORG] Sent: Wednesday, May 19, 2004 8:22 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Alternative fixative users (Prefer) Hi, after many years as a Prefer fixative user our pathologists are telling us that we have to go back to formalin. This came about after a letter to the editor appeared in the Journal of Surgical Pathology. Has anyone out there been approached by their pathologists to switch back? Mary Bryhan _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nlouisea <@t> gis.net Tue May 18 11:13:18 2004 From: nlouisea <@t> gis.net (Nancy Adams) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] evaluation of non pathologist grossing tissue Message-ID: <004f01c43cf3$0971f800$7b20fea9@eds-desk> Good morning Does anyone have good documenting paper work to evaluate non pathologist (also not a PA) grossing tissue? We have a daily check sheet that the pathologist reading the cases signs but I'm looking for something better to answer the CAP question ANP.11640 about regular and periodic evaluation. Thanks Nancy Adams Falmouth Hospital Falmouth, MA From pzeitlow <@t> bbpllab.com Wed May 19 11:20:18 2004 From: pzeitlow <@t> bbpllab.com (Pat Zeitlow) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Varicella Zoster by in situ Message-ID: <813FB33DA405334F947F8BFC6EBD0B2AE673@bbplsrv1.bbpl> Does anyone have any ideas where I could send a block to for testing Varicella zoster by in situ hybridization. I have a request and very little tissue to provide. I welcome any suggestions! (Thanks to Martha at Wake Forest for referring me to the histonet!) Pat Department of Molecular Pathology Boyce & Bynum Laboratories, P.C. Columbia, Missouri From rocan <@t> mac.com Wed May 19 11:17:09 2004 From: rocan <@t> mac.com (rocan@mac.com) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] HIF 1 alpha In-Reply-To: References: Message-ID: Garry, What is the source of your antibody. Most HIF-1a antibodies are very, very poor and difficult. On May 19, 2004, at 8:32 AM, Garry Ashton wrote: > Dear all, > I'm having problems with an antibody we are using. The antibody is HIF > 1alpha. > We have had some good staining, and then some poor results. > It was mentioned that the age of the sections could have something to > do > with it. > Has any experienced this before, or know of any other reason. > The antibody has been stored as instructed and is well within date. > Many thanks for any help in advance. > Garry > > > PICR > UK > > -------------------------------------------------------- > > > This email is confidential and intended solely for the use of the > person(s) ('the intended recipient') to whom it was addressed. Any > views or opinions presented are solely those of the author and do not > necessarily represent those of the Paterson Institute for Cancer > Research or the Christie Hospital NHS Trust. It may contain > information that is privileged & confidential within the meaning of > applicable law. Accordingly any dissemination, distribution, copying, > or other use of this message, or any of its contents, by any person > other than the intended recipient may constitute a breach of civil or > criminal law and is strictly prohibited. If you are NOT the intended > recipient please contact the sender and dispose of this e-mail as soon > as possible. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ploykasek <@t> phenopath.com Wed May 19 11:31:20 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] p16INK4a In-Reply-To: <328CBAE62F31C642B422970E879DFADC093CB9@pcwex01.meriter.com> Message-ID: We have used several different clones of p16ink4. My understanding is the polyclonal from Pharingen is no longer available. A really good clone out of Europe is only available through Dakocytomation in their CINtek kit, this is clone E6H4. We have tired the kit, and got good results. Currently we are using the following: Antibody: p16INK4a Clone: 16PO4 (akaJC2) Supplier: Lab Vision Pretreatment: Steam for 40? in Tris, pH10 Titer: 1:200 Detection: Envision+ Localization: both nuclear & cytoplasmic Hope this info gets you started. Good luck with your staining. Patti Loykasek PhenoPath Laboratories Seattle, WA > The Pathologists I work for have requested that I purchase p16INK4a. I'd like > to get an idea of which company most people order from and whether you use the > monoclonal or polyclonal antibody. > > Thanks, > > Jean Taylor > Immuno Tech > GML > Madison, WI > > -----Original Message----- > From: Margaret Blount [mailto:mab70@medschl.cam.ac.uk] > Sent: Wednesday, May 19, 2004 09:58 > To: 'Smith, Emily'; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Background Staining > > > Dear Emily, > > That's a strange one. Have you titrated your secondary antibody > sufficiently? what blocking solution do you use? Have the slides > inadvertently dried out at any stage of the procedure? > > These are a few thoughts that spring to mind. The other thing that I would > look at would be to try charged slides from another supplier, or try > Polysine or APES slides for comparison on indeed another batch of slides. > That's my 2 penny worth - maybe someone else will have something more > helpful to say and no doubt you have thought of some or all of these ideas > yourself. > > Good luck. > > Regards > > Margaret > > Margaret Blount > Chief Technician > Clinical Biochemistry > University of Cambridge > Addenbrooke's Hospital > Hills Road > Cambridge > CB2 2QR > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Smith, > Emily > Sent: Wednesday, May 19, 2004 3:42 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Background Staining > > > Hello All, > In the course of a recent experiment, I saw strong background staining > on charged slides with frozen cells in OCT (optimal cutting temperature) > when treated with streptavidin-HRP. > The staining appears over the entire glass surface not only where the > cells and OCT were. This all-over-staining also appears on clean > (without OCT on them) slides that are processed with the OCT slides. I > have tried extensive water washes and the results remain the same. No > primary antibody is added and the results are the same. > Have you seen this before? Do you have any suggestions? > Thank you for your help. > ~Emily > > Emily A. Smith > CuraGen Corporation > > > 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 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 > > _______________________________________________ > 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 Wed May 19 11:18:52 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Tunel Message-ID: I'm looking for some tips, info on tunel assays, specifically the kit from Promega - "dead end" tunel kit. Thanks for the help. Patti Loykasek From emry <@t> u.washington.edu Wed May 19 12:36:29 2004 From: emry <@t> u.washington.edu (P. Emry) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] H&E stain problems In-Reply-To: References: Message-ID: Ok, I have this pain that starts behind my right eye. Let me get this straight... I don't have to do 5 mins in clearing x3 100etoh x3 95etohx3 just 20-30 vigorus dips? Am I getting this? then 20mins in whosoever's-hemotox, 15 mins in running water. 20-30 vigorus dips in 70etoh, 95x3,100ethox3,clearingx3 then slipcover. Where does the acid alcohol come in and for howlong? How did I miss this after all these years. Thanks, and I knew Frank Sinatra....just thought I'd throw that in for the music lovers here. Trisha Seattle On Wed, 19 May 2004, Marshall Terry Dr, Consultant Histopathologist wrote: > Close relative:-) > > 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: Connie McManus [mailto:convmcm@cc.usu.edu] > Sent: 19 May 2004 14:49 > To: 'Gary Gill'; Marshall Terry Dr, Consultant Histopathologist; 'Petia > P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu > Cc: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] H&E stain problems > > > Gary > > Is Gill a relative or yours??? I don't give a hoot who made what when. > I like Harris. I don't believe Gill has made any significant > improvements over the old dead white guy's hematoxylin. > > BTW, classical music may have been composed by dead white guys, but I > don't hear ANYTHING being composed today that has the depth, complexity > and color of those great composers. I've studied piano from since I was > 4 yrs old, and I've studied the organ (I've played as a church > organist)for almost 20 years. I know music like the back of my hand > and I love ALL music (except gangstah stuff). SO DON'T try to tell me > about what music is living and worth my while. > > Dead white Europeans. Yeah, right. I'd like to see someone between 8 and > 14 in this generation compose music like Felix Mendelsohnn's or > Mozart's. > > Connie McManus > Utah Veterinary Diagnostics Laboratory > Utah State University > Logan, UT > Phone: 435/797-1891 > fax: 435/797-2805 > > > -----Original Message----- > From: Gary Gill [mailto:garygill@dcla.com] > Sent: Tuesday, May 18, 2004 10:04 AM > To: 'Connie McManus'; 'Marshall Terry Dr,Consultant Histopathologist'; > 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu > Cc: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] H&E stain problems > > You may have heard that classical music was composed by dead white > Europeans. Well, Harris hematoxylin was composed by a dead white > American > (physician at Jefferson Hospital in 1904). Gill hematoxylin was > composed by > a live white American. So if you want to liven things up, go with > Gill's! > > Gary Gill (one and the same) > > PS -- No royalties involved, thanks to bad advice in 1972 from corporate > counsel for Johns Hopkins Medical School. > > -----Original Message----- > From: Connie McManus [mailto:convmcm@cc.usu.edu] > Sent: Tuesday, May 18, 2004 10:41 AM > To: 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P > Stefanova'; > 'Megan Kear'; histonet@lists.utsouthwestern.edu > Cc: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] H&E stain problems > > > Wow. What a lot of interesting comments!! > > I agree with Terry re the agitation. When I watch the stainer do those > dips > (I can program how many, but NOT the briskness), I wonder if you could > even > call it agitation. My hand dips are very brisk. Also, I don't bother > letting the slides stay in the alcohols for 1 minute or 2, I give the > slides > about 20 -30 good brisk dips in each solution, then the timed rinses & > staining. This has always been far more satisfactory to me than those > sllllooooowwwww dips from the machine. > > As for the kind of hematoxylin, someone suggested I throw out the Harris > and > do Gills III. I've tried Gill III before and I much prefer the Harris. > So > it's just a matter of personal preference on that... AND what your > pathologist likes *G* > > Everyone having an nice Tuesday??? *g* > > Connie McManus > Utah Veterinary Diagnostics Laboratory > Utah State University > Logan, UT > Phone: 435/797-1891 > fax: 435/797-2805 > > > -----Original Message----- > From: Marshall Terry Dr, Consultant Histopathologist > [mailto:Terry.Marshall@rothgen.nhs.uk] > Sent: Tuesday, May 18, 2004 7:35 AM > To: Connie McManus; Petia P Stefanova; Megan Kear; > histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] H&E stain problems > > Connie remarks: > > "In truth, I prefer my hand stained sections better than when they're > stained automatically." > > When I first saw what I call x-y stainers, I thought that we had in > this, > something that reproduced hand staining. Not so. What I think is lacking > is > the brisk agitation necessary to break down the interface between old > and > new solution. I'm not so sure about the rinsing either. > > 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: Connie McManus [mailto:convmcm@cc.usu.edu] > Sent: 18 May 2004 15:09 > To: 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] H&E stain problems > > > We use almost the exact same protocol... we use Surgipath Harris, but we > prepare eosin in-house. One thing I am amazed at in this protocol is > the > length of time in the acid alcohol. Do you use an autostainer? We have > a > Leica. The time is set to 1 second in the acid ETOH and > sometimes the sections are almost too differentiated. I can't imagine > 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the > slides > in and quickly put them in running water. I must have a very strong > solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand > stained > sections better than when they're stained automatically. > > Just wondering and blabbering (hey, it's Tuesday, what do you expect??) > > Connie McManus > Utah Veterinary Diagnostics Laboratory > Utah State University > Logan, UT > Phone: 435/797-1891 > fax: 435/797-2805 > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P > Stefanova > Sent: Monday, May 17, 2004 6:45 AM > To: Megan Kear; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] H&E stain problems > > Hi, > > I use Harris's hematoxylin which is also regressive and purchase my > hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very > good H&E staining with this protocol. > > REAGENT TIME > Xylene 3 min. > Xylene 3 min. > Abs. alc. 2 min. > Abs. alc. 2 min > 95% alc. 2 min > 80% alc. 2 min > Wash /tap water/ 30 sec. > Hematoxylin 8 min. > Wash /tap water/ 2 min. > Acid alcohol 6 sec. > Wash /tap water/ 2 min. > Wash /tap water/ 5 min > 80% alc. 30 sec. > Eosin 15 sec. > 95% alc. 10 sec. > Abs. alc. 30 sec. > Abs. alc. 30 sec. > Xylene 1 min. > Xylene 1 min. > Xylene Exit > > Hope it helps! > Petia > > > _______________________________________________ > 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 Charlene.Henry <@t> STJUDE.ORG Wed May 19 12:28:05 2004 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Tunel Message-ID: <5CB39BCA5724F349BCB748675C6CA1A2315834@SJMEMXMB02.stjude.sjcrh.local> We currently use the Promega "DeadEnd Colorimetric TUNEL System" kit with excellent results. I have also programmed it on my Ventana Discovery instrument; so it is completely automated. We follow the protocol included in the kit and the only reagent not included is the DNase which we use at a 1:20 dilution for 30 minutes at 37 degrees C. We get our "DNA POLIDNase 1" from Invitogen cat # 18162016. Instead of the 4% paraformaldehyde, we use 37% formaldehyde to make a 10% solution in PBS. Hope this helps. Charlene Henry, HT QIHC Histology/IHC Section Head Department of Pathology St. Jude Children's Research Hospital 332 North Lauderdale Memphis, TN 38105 Phone: 901-495-3349 FAX: 901-495-3100 -----Original Message----- From: Patti Loykasek [mailto:ploykasek@phenopath.com] Sent: Wednesday, May 19, 2004 11:19 AM To: histonet Subject: [Histonet] Tunel I'm looking for some tips, info on tunel assays, specifically the kit from Promega - "dead end" tunel kit. Thanks for the help. Patti Loykasek _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LuckG <@t> empirehealth.org Wed May 19 13:08:03 2004 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:23:00 2005 Subject: FW: [Histonet] Alternative fixative users (Prefer) Message-ID: -----Original Message----- From: Luck, Greg D. Sent: Wednesday, May 19, 2004 11:04 AM To: 'histonet@pathology.swmed.edu' Subject: RE: [Histonet] Alternative fixative users (Prefer) Mary, We have gone down this path already (about two years ago). I can offer the solution that I came up with and has proved very effective and "nearly" flawless. It allows for the necessary preservation of tissues that are either non-reactive to certain antibodies (eg. WT-1, TTF-1, E-cadherin, ER and PR; ie. these are the ones we have been unable to work up on Prefer fixed tissues) while enjoying the vastly superior results on the overwhelming number of antibodies done on the Prefer fixed tissues. In addition it keeps the more pleasant working environment with Prefer as the dominant fixative. I have included a portion of my "Histology Specimen Handling" policy at the end of my e-mail. Of particular interest to you will be the "Fixation" and the "Labeling Cassettes and Slides" sections. I urge you to give me a call and I can give you some more details on answering your pathologists concerns and in steps of implementation after I get a clear picture of your operational circumstances. It obviously wouldn't work in every situation but might be a good fit for you or at least a trial for feasibility. Greg Luck Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org Fixation: a) Specimens are "routinely" fixed in Prefer. Fixative should be added as soon as possible; exceptions being specimens which are submitted for frozen section, or for immediate exam and consultation or at the request of the grossing pathologist. b) All breast specimens and some selected tissue block cassettes are fixed in 10% N.B.F. (eg. neoplasms of the ovary, lung and lymphomas) as directed by the grossing pathologist. Labeling Cassettes: a) Green cassettes and slides are prepared for routine surgical specimens fixed in Prefer and yellow cassettes and slides for those fixed in 10% N.B.F. b) White cassettes are prepared for cytology specimens and bone marrows. c) Blue cassettes are prepared for autopsy specimens. -----Original Message----- From: Mary Bryhan [mailto:mbryhan@NORTHERNHEALTH.ORG] Sent: Wednesday, May 19, 2004 8:22 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Alternative fixative users (Prefer) Hi, after many years as a Prefer fixative user our pathologists are telling us that we have to go back to formalin. This came about after a letter to the editor appeared in the Journal of Surgical Pathology. Has anyone out there been approached by their pathologists to switch back? Mary Bryhan _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LuckG <@t> empirehealth.org Wed May 19 13:04:11 2004 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Alternative fixative users (Prefer) Message-ID: Mary, We have gone down this path already (about two years ago). I can offer the solution that I came up with and has proved very effective and "nearly" flawless. It allows for the necessary preservation of tissues that are either non-reactive to certain antibodies (eg. WT-1, TTF-1, E-cadherin, ER and PR; ie. these are the ones we have been unable to work up on Prefer fixed tissues) while enjoying the vastly superior results on the overwhelming number of antibodies done on the Prefer fixed tissues. In addition it keeps the more pleasant working environment with Prefer as the dominant fixative. I have included a portion of my "Histology Specimen Handling" policy at the end of my e-mail. Of particular interest to you will be the "Fixation" and the "Labeling Cassettes and Slides" sections. I urge you to give me a call and I can give you some more details on answering your pathologists concerns and in steps of implementation after I get a clear picture of your operational circumstances. It obviously wouldn't work in every situation but might be a good fit for you or at least a trial for feasibility. Greg Luck Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org Fixation: a) Specimens are "routinely" fixed in Prefer. Fixative should be added as soon as possible; exceptions being specimens which are submitted for frozen section, or for immediate exam and consultation or at the request of the grossing pathologist. b) All breast specimens and some selected tissue block cassettes are fixed in 10% N.B.F. (eg. neoplasms of the ovary, lung and lymphomas) as directed by the grossing pathologist. Labeling Cassettes: a) Green cassettes and slides are prepared for routine surgical specimens fixed in Prefer and yellow cassettes and slides for those fixed in 10% N.B.F. b) White cassettes are prepared for cytology specimens and bone marrows. c) Blue cassettes are prepared for autopsy specimens. -----Original Message----- From: Mary Bryhan [mailto:mbryhan@NORTHERNHEALTH.ORG] Sent: Wednesday, May 19, 2004 8:22 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Alternative fixative users (Prefer) Hi, after many years as a Prefer fixative user our pathologists are telling us that we have to go back to formalin. This came about after a letter to the editor appeared in the Journal of Surgical Pathology. Has anyone out there been approached by their pathologists to switch back? Mary Bryhan _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bwhitaker <@t> brownpathology.com Wed May 19 13:15:57 2004 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Background Staining In-Reply-To: <6C21D947F7189448AB447C57BF2992B4025F16C5@mius.medlan.cam.ac.uk> Message-ID: <000901c43dcd$54d6cd90$3601a8c0@brownpathology.net> Hi! When I was a rep for an immuno vendor, I did technical training on the cap gap method, anyway, we frequently saw the phenomenon you describe of staining on blank slides that we occasionally used to pair with a tissue-bearing slide to form the cap gap. I think it's a function of a strong charge. BTW, we didn't see this as much on slides that underwent HIER or digestion, but that doesn't help your FS protocol. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Margaret Blount Sent: Wednesday, May 19, 2004 9:58 AM To: 'Smith, Emily'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Background Staining Dear Emily, That's a strange one. Have you titrated your secondary antibody sufficiently? what blocking solution do you use? Have the slides inadvertently dried out at any stage of the procedure? These are a few thoughts that spring to mind. The other thing that I would look at would be to try charged slides from another supplier, or try Polysine or APES slides for comparison on indeed another batch of slides. That's my 2 penny worth - maybe someone else will have something more helpful to say and no doubt you have thought of some or all of these ideas yourself. Good luck. Regards Margaret Margaret Blount Chief Technician Clinical Biochemistry University of Cambridge Addenbrooke's Hospital Hills Road Cambridge CB2 2QR -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Smith, Emily Sent: Wednesday, May 19, 2004 3:42 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Background Staining Hello All, In the course of a recent experiment, I saw strong background staining on charged slides with frozen cells in OCT (optimal cutting temperature) when treated with streptavidin-HRP. The staining appears over the entire glass surface not only where the cells and OCT were. This all-over-staining also appears on clean (without OCT on them) slides that are processed with the OCT slides. I have tried extensive water washes and the results remain the same. No primary antibody is added and the results are the same. Have you seen this before? Do you have any suggestions? Thank you for your help. ~Emily Emily A. Smith CuraGen Corporation 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 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From STapper <@t> slhduluth.com Wed May 19 13:23:06 2004 From: STapper <@t> slhduluth.com (Tapper, Sheila) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Dako autostainer vial storage Message-ID: <3BFBBD68413CB443A7125A63EC0ACD1D6716DC@slhw2smail01.slhdomain.com> I am just wondering if anyone out there has found a tray that works well with the reagent vials for the Autostainer - other than the racks that are used on the machine? A rack that would allow room for a label would be preferred - I can't seem to find anything in the plethora of catalogs that come through the lab!!! I know there are many users out there - how are you organizing your antibodies in the refrigerator? Sheila Tapper HT(ASCP) stapper@slhduluth.com St. Luke's Hospital Duluth, MN CONFIDENTIAL: The confidential information accompanying this transmission may contain protected health information and is legally privileged under state and federal law. This information is intended only for the use of the individual to which it is addressed. The recipient or person responsible for delivering this information is prohibited by law from disclosing this information without proper authorization to any other party, unless required to do so by law or regulation. If you are not the intended recipient, you are hereby notified that any review, 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. No response indicates that the information was received by the appropriate authorized party. Warning: Although St. Luke's has taken reasonable precautions to ensure no viruses are present in this email, St. Luke's cannot accept responsibility for any loss or damage arising from the use of this email or attachments. From RossS <@t> BaylorHealth.edu Wed May 19 13:32:15 2004 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Dako autostainer vial storage Message-ID: We always ordered extra of the racks that come with the machine. When we had to Shandon Cadenza I made a holder for the small vials by finding an appropriately sized container and filling it with paraffin. Then when it was almost cool, I used a vial to make holes in the paraffin. The result was a perfectly sized tray. It didn't look pretty after a few months, but it was used for about a year. You could do something similar. I'm sure there will be other suggestions as well. 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 Tapper, Sheila Sent: Wednesday, May 19, 2004 1:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Dako autostainer vial storage I am just wondering if anyone out there has found a tray that works well with the reagent vials for the Autostainer - other than the racks that are used on the machine? A rack that would allow room for a label would be preferred - I can't seem to find anything in the plethora of catalogs that come through the lab!!! I know there are many users out there - how are you organizing your antibodies in the refrigerator? Sheila Tapper HT(ASCP) stapper@slhduluth.com St. Luke's Hospital Duluth, MN CONFIDENTIAL: The confidential information accompanying this transmission may contain protected health information and is legally privileged under state and federal law. This information is intended only for the use of the individual to which it is addressed. The recipient or person responsible for delivering this information is prohibited by law from disclosing this information without proper authorization to any other party, unless required to do so by law or regulation. If you are not the intended recipient, you are hereby notified that any review, 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. No response indicates that the information was received by the appropriate authorized party. Warning: Although St. Luke's has taken reasonable precautions to ensure no viruses are present in this email, St. Luke's cannot accept responsibility for any loss or damage arising from the use of this email or attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From mkundu <@t> purdue.edu Wed May 19 13:39:29 2004 From: mkundu <@t> purdue.edu (Moumita Kundu) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Dako autostainer vial storage References: <3BFBBD68413CB443A7125A63EC0ACD1D6716DC@slhw2smail01.slhdomain.com> Message-ID: <001101c43dd0$9e548e50$4496d280@agriculture.purdue.edu> There are racks in fisher catalog whcih are perfect fit for the Dako autostainer vials ,they are cheaper than the dakos.Let me know if youneed the catalog number. ----- Original Message ----- From: "Tapper, Sheila" To: Sent: Wednesday, May 19, 2004 1:23 PM Subject: [Histonet] Dako autostainer vial storage > I am just wondering if anyone out there has found a tray that works well > with the reagent vials for the Autostainer - other than the racks that > are used on the machine? A rack that would allow room for a label would > be preferred - I can't seem to find anything in the plethora of catalogs > that come through the lab!!! I know there are many users out there - > how are you organizing your antibodies in the refrigerator? > > Sheila Tapper HT(ASCP) > stapper@slhduluth.com > St. Luke's Hospital > Duluth, MN > > > > CONFIDENTIAL: > The confidential information accompanying this transmission may contain > protected health information and is legally privileged under state and > federal law. This information is intended only for the use of the > individual to which it is addressed. The recipient or person > responsible for delivering this information is prohibited by law from > disclosing this information without proper authorization to any other > party, unless required to do so by law or regulation. If you are not > the intended recipient, you are hereby notified that any review, > 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. No response indicates that the > information was received by the appropriate authorized party. > > Warning: Although St. Luke's has taken reasonable precautions to ensure > no viruses are present in this email, St. Luke's cannot accept > responsibility for any loss or damage arising from the use of this > email or attachments. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From akwilliams75 <@t> hotmail.com Wed May 19 13:35:15 2004 From: akwilliams75 <@t> hotmail.com (kevin williams) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Slides & blocks - Biohazard? Message-ID: Prions, are not regulations and if these are going into land fill, surly the odd tampon has more prions than shavings. >From: Jackie.O'Connor@abbott.com >To: "kevin williams" >CC: histonet@pathology.swmed.edu, Rcartun@harthosp.org >Subject: RE: [Histonet] Slides & blocks - Biohazard? >Date: Tue, 18 May 2004 09:49:55 -0500 > >Prions. > > > > > > >"kevin williams" >Sent by: histonet-bounces@lists.utsouthwestern.edu >05/18/2004 09:31 AM > > > To: Rcartun@harthosp.org, histonet@pathology.swmed.edu > cc: > Subject: RE: [Histonet] Slides & blocks - Biohazard? > > > > Dear Richard: > > I have been told that shavings are not Biohazard, because they have > been processed, so how can a single "shaving" on a slide be considered > biohazard? > > Yours faithfully, > > A. Kevin Williams > >From: "Richard Cartun" > >To: > >Subject: [Histonet] Slides & blocks - Biohazard? > >Date: Mon, 17 May 2004 14:22:21 -0400 > > > >Are glass microscopic slides and paraffin tissue blocks considered > >"biohazards"? We occasionally receive either unstained slides or a > >paraffin tissue block in a "Biohazard" bag from another institution > for > >special testing. Technically speaking, the only personnel allowed to > >handle those materials would be those individuals wearing gloves. > > > >Richard Cartun > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________________________ __ > > [1]Stop worrying about overloading your inbox - get MSN Hotmail Extra > Storage! > >References > > 1. http://g.msn.com/8HMBENUS/2737??PS=47575 >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _________________________________________________________________ [1]Watch LIVE baseball games on your computer with MLB.TV, included with MSN Premium! References 1. http://g.msn.com/8HMAENUS/2740??PS=47575 From ploykasek <@t> phenopath.com Wed May 19 13:35:46 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Alternative fixative users (Prefer) In-Reply-To: Message-ID: I would caution any using Prefer to consider if they will be doing any rna/dna testing now or perhaps in the future on the tissue. We have had poor dna FISH results with it. Soon I'll be trying some pcr assays, and will let you know how that goes. Patti Loykasek PhenoPath Laboratories Seattle, WA > Hi, after many years as a Prefer fixative user our pathologists are telling > us that we have to go back to formalin. This came about after a letter to the > editor appeared in the Journal of Surgical Pathology. > > Has anyone out there been approached by their pathologists to switch back? > > Mary Bryhan > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From shive003 <@t> umn.edu Wed May 19 13:54:11 2004 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Dako autostainer vial storage References: <3BFBBD68413CB443A7125A63EC0ACD1D6716DC@slhw2smail01.slhdomain.com> Message-ID: <00b701c43dd2$ac9503d0$78065486@vdl220FAC> I don't know where you could find the right size rack for Dako vials, other than that company. I buy extra Dako racks and use them for storing vials. Diluted abs are stored alphabetically in several racks. Detection reagents in another one. Jan Shivers U of MN Vet Diag Lab St. Paul, MN ----- Original Message ----- From: "Tapper, Sheila" To: Sent: Wednesday, May 19, 2004 1:23 PM Subject: [Histonet] Dako autostainer vial storage I am just wondering if anyone out there has found a tray that works well with the reagent vials for the Autostainer - other than the racks that are used on the machine? A rack that would allow room for a label would be preferred - I can't seem to find anything in the plethora of catalogs that come through the lab!!! I know there are many users out there - how are you organizing your antibodies in the refrigerator? Sheila Tapper HT(ASCP) stapper@slhduluth.com St. Luke's Hospital Duluth, MN CONFIDENTIAL: The confidential information accompanying this transmission may contain protected health information and is legally privileged under state and federal law. This information is intended only for the use of the individual to which it is addressed. The recipient or person responsible for delivering this information is prohibited by law from disclosing this information without proper authorization to any other party, unless required to do so by law or regulation. If you are not the intended recipient, you are hereby notified that any review, 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. No response indicates that the information was received by the appropriate authorized party. Warning: Although St. Luke's has taken reasonable precautions to ensure no viruses are present in this email, St. Luke's cannot accept responsibility for any loss or damage arising from the use of this email or attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mayers <@t> newnanhospital.org Wed May 19 14:07:25 2004 From: mayers <@t> newnanhospital.org (mayers@newnanhospital.org) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] (no subject) Message-ID: We are trying to locate 1/2 and 3/4 inch metal tampers for embedding. Anyone have a source? Thanks Mike Ayers BA HT(ASCP) Supervisor Histology Newnan Hospital 60 Hospital Road Newnan, Georgia 30263 Office 770-304-4065 Fax 770-253-2570 From carl.hobbs <@t> kcl.ac.uk Wed May 19 14:08:38 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Charged slides Message-ID: <001a01c43dd4$b161e340$94509b51@home> Anyone know what the difference is between the "charged" slides we buy/make? Certainly, the price we pay must be directly proportional to the extent of charging lol but, OK I can use PLL, APES, chrome alum/gelatin or Superfrost plus!! Are the latter charged in a different way? Yes, I hear they are electrostatically charged but....how? is there a charge density/ property difference? Be grateful for clarification. NB I have my own views on the efficacy of the ones mentioned but would be very interested in others' experience. --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.684 / Virus Database: 446 - Release Date: 13/05/2004 From lleach <@t> uic.edu Wed May 19 14:13:12 2004 From: lleach <@t> uic.edu (Lu Leach) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] racks Message-ID: <6.0.1.1.2.20040519141110.02400470@tigger.uic.edu> Newcomer Supply has racks that work with the vials from DAKO. The order number is #5801 A, they hold 40 vials and cost $72. Contact Newcomer at 800-383-7799 and tell Marcia "hello" from me. From carl.hobbs <@t> kcl.ac.uk Wed May 19 14:34:50 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] RE: H&E stain problems Message-ID: <002901c43dd8$5a7083d0$94509b51@home> A gem that has been circulating since my eary days of the 70s. It goes quiet for a few years then..suprise to me , resurfaces. I remember being squashed by Wallington in a AMM meeting in London for not only suggesting the use of DPX over Canada Balsam for routine mounting but also for replacing the best "PORT" of the Hx staining solutions ( Ehrlich's) , with Harris',for routine staining. After spending several years as an EQUAS assessor( UK, Southeast) there is no doubt, to me, that a hand-processed H&E ( using a well- ripened Ehrlich's or a well- made Harris' , by an experienced hand) is far superior to a machine-produced result. They most certainly can produce the best photomicrographs( lets not get into the different fixing fluids that enhance the quality lol). However, for routine diagnostic work I am convinced that these are not neccessary. Surely, for the vast majority of sections , we want a fast, consistent, result, with the minimum amount of "craft" involved? Sure, the "progressive" Hxs don't give the best staining but, they more assuredly give the "good enough " staining for such throughput required by most routine diagnostic labs? --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.684 / Virus Database: 446 - Release Date: 13/05/2004 From mbecker <@t> pathlabinc.com Wed May 19 14:58:55 2004 From: mbecker <@t> pathlabinc.com (Michelle Becker) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] (no subject) In-Reply-To: Message-ID: Mike, We purchased ours from Cardinal,formerly Allegiance. 1-800-964-5227 Cat # 1551 Large tamper Cat # 1552 Small tamper (1/2") Michele Becker, HTL(ASCP) Histology Manager Path Lab Inc., A Lab Corp Company 195 Hanover Street Portsmouth, NH 03801 (603)431-2310 x4220 mbecker@pathlabinc.com (603)431-3741 Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of mayers@newnanhospital.org Sent: Wednesday, May 19, 2004 2:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) We are trying to locate 1/2 and 3/4 inch metal tampers for embedding. Anyone have a source? Thanks Mike Ayers BA HT(ASCP) Supervisor Histology Newnan Hospital 60 Hospital Road Newnan, Georgia 30263 Office 770-304-4065 Fax 770-253-2570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cgfields <@t> lexhealth.org Wed May 19 15:04:40 2004 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] (no subject) Message-ID: Cardinal Health Cat.#____ Tissue-Tek Tamper, sm 1552 3/pkg $15.40 Tissue-Tek Tamper, lg 1551 3/pkg $19.40 Old prices but they have them. Carole Fields Lex Med Ctn W.Columbia,SC -----Original Message----- From: mayers@newnanhospital.org [mailto:mayers@newnanhospital.org] Sent: Wednesday, May 19, 2004 3:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) We are trying to locate 1/2 and 3/4 inch metal tampers for embedding. Anyone have a source? Thanks Mike Ayers BA HT(ASCP) Supervisor Histology Newnan Hospital 60 Hospital Road Newnan, Georgia 30263 Office 770-304-4065 Fax 770-253-2570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From Apeters <@t> chsd.org Wed May 19 15:11:11 2004 From: Apeters <@t> chsd.org (Peters, Anne) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Cryostat Problems Message-ID: <00D7298B3825D511BF540008C75F8A3C8BB592@EXCLUSTER.chsd.org> Has anyone had any problems with the peltier on their Microm HM 505E? We're on our 3rd board. Do we just have a lemon? Tx, Anne From madaryhtl <@t> juno.com Wed May 19 15:40:16 2004 From: madaryhtl <@t> juno.com (madaryhtl@juno.com) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Tunel Message-ID: <20040519.134039.22945.25054@webmail14.lax.untd.com> We use the Cat #S7200 Apoptag kit from Serologicals, it works great. www.intergen.com From Ronnie_Houston <@t> bshsi.com Wed May 19 15:55:22 2004 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Slides & blocks - Biohazard? Message-ID: <530361BF03351B4CAE5270A05D3037B50367328F@bsrexms01.BSHSIR.COM> Kevin, You should be reviewing documents from your home base, UK, before commenting about prions. Unless infected tissue is post-fixed in formic acid for 1 hour, and then further fixed in formalin, it is still considered infectious. Shavings from a CJD case must then be collected, bagged and incinerated. However, once a section has been coverslipped and sealed, and then the slide wiped with a recommended disinfectant, eg sodium hypochlorite (20,000ppm available chlorine), it is considered safe and should be treated as you would a normal slide. Incidentally, tissue and fluids from the female reproductive tract are not classified as being infectious; although placenta is classed as being of low infectivity and should be incinerated. Ronnie Ronnie Houston Director of Anatomic Pathology Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 (804) 287 7906 - fax ronnie_houston@bshsi.com -----Original Message----- From: kevin williams [mailto:akwilliams75@hotmail.com] Sent: Wednesday, May 19, 2004 2:35 PM To: Jackie.O'Connor@abbott.com; histonet@pathology.swmed.edu Subject: RE: [Histonet] Slides & blocks - Biohazard? Prions, are not regulations and if these are going into land fill, surly the odd tampon has more prions than shavings. >From: Jackie.O'Connor@abbott.com >To: "kevin williams" >CC: histonet@pathology.swmed.edu, Rcartun@harthosp.org >Subject: RE: [Histonet] Slides & blocks - Biohazard? >Date: Tue, 18 May 2004 09:49:55 -0500 > >Prions. > > > > > > >"kevin williams" >Sent by: histonet-bounces@lists.utsouthwestern.edu >05/18/2004 09:31 AM > > > To: Rcartun@harthosp.org, histonet@pathology.swmed.edu > cc: > Subject: RE: [Histonet] Slides & blocks - Biohazard? > > > > Dear Richard: > > I have been told that shavings are not Biohazard, because they have > been processed, so how can a single "shaving" on a slide be considered > biohazard? > > Yours faithfully, > > A. Kevin Williams > >From: "Richard Cartun" > >To: > >Subject: [Histonet] Slides & blocks - Biohazard? > >Date: Mon, 17 May 2004 14:22:21 -0400 > > > >Are glass microscopic slides and paraffin tissue blocks considered > >"biohazards"? We occasionally receive either unstained slides or a > >paraffin tissue block in a "Biohazard" bag from another institution > for > >special testing. Technically speaking, the only personnel allowed to > >handle those materials would be those individuals wearing gloves. > > > >Richard Cartun > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________________________ __ > > [1]Stop worrying about overloading your inbox - get MSN Hotmail Extra > Storage! > >References > > 1. http://g.msn.com/8HMBENUS/2737??PS=47575 >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _________________________________________________________________ [1]Watch LIVE baseball games on your computer with MLB.TV, included with MSN Premium! References 1. http://g.msn.com/8HMAENUS/2740??PS=47575 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone at 410-442-3250, and permanently delete the original e-mail, attachment(s), and any copies. From lagehan <@t> hotmail.com Wed May 19 20:32:20 2004 From: lagehan <@t> hotmail.com (LORALEE GEHAN) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] CJD DECONTAMINATION PROCEDURE Message-ID: Your probably going to have to process by hand. Formic acid for one hour, the formalin overnight. Then just routinely process by hand. Everything in disposable containers. Use 5N sodium hydroxide to all of the processing solutions to "neutrilize" and have your bioharzard people come in a get it. Wet down everything with 5N sodium hydroxide and let it sit for a bit. Then wipe it off with wet paper towels. Everything should be red bagged and incinerated. I used to work for the Brain Bank for AD research. We used to have fits about Dr's performing biopsy's on patients suspect of CJD, because CJD doesn't show up everywhere in the brain and the chances that they are going to get the peice that shows evidence of CJ is slim. And I don't know of any treatment for CJD so, the point of the biopsy? Hopefully the surgical team will be using disposables and wetting everything in the OR down with 5N sodium hydroxide before continueing to the next patient. You should also store the blocks and slides seperate from everything else. We had a speical area devoted to such cases. Any other questions feel free to ask. Hope that helps a little. We didn't get any hazard pay!! Loralee Gehan, HTL >From: "Connie McManus" >To: , >Subject: RE: [Histonet] CJD DECONTAMINATION PROCEDURE >Date: Wed, 19 May 2004 08:03:16 -0600 > >I don't deal with CJD, but I do a lot of CWD (Chronic Wasting Disease) >and Scrapie, which are prion diseases in dear/elk and sheep >(respectively). CWD and Scrapie had been associated with humans for >hundreds of years with no known transmission of the disease from animal >to human. HOWEVER, since prions are not neutralized by fixation of any >kind, not killed by autoclaving, this makes them a biohazard in >processed tissues. > >If I worked with CJD, I would ... >1 set up a place that is completely separate from the rest of the lab. >Barrier off a section of your lab, use another room (preferable) > >2 use disposable bunny suits, shoe covers, face masks, eye wear and >DOUBLE GLOVE >3 put stickie mats on the floor completely surrounding the cutting >area, > >4 put ALL paraffin shavings in a biohaz bag, > >5 use paper towels to clean off the microtome and throw those in the >biohaz bag > >6. incinerate the bunny suit, goggles, gloves, face maskes, etc with the >paraffin shavings. > >7. Ask for a really big pay raise. Claim it's hazard pay. > >Have fun. >Connie McManus >Utah Veterinary Diagnostics Laboratory >Utah State University >Logan, UT >Phone: 435/797-1891 >fax: 435/797-2805 > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >srishan@mail.holyname.org >Sent: Tuesday, May 18, 2004 9:59 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] CJD DECONTAMINATION PROCEDURE > > > > > >Hello everyone, > >This question might have been posted before, but, Histology Department >is >expecting a brain biopsy from a possible CJD patient today. The >specimen >will be coming down in formalin from the O.R. It will be kept in >formalin >for 2-7 days and will be transferred to formic acid for the required >time. >Then the specimen will be transferred to formalin before processing for >another 2 days. Has anyone dealt with this procedure before. These are >the >questions asked by the hstology department. > >How do you decontaminate the processor, microtome, stainer and >coverslipper? >What kind of protective gear apart from standard precaution has to be >used? >How about the shaving of the tissues? >How do you dispose the formalin from the original formalin container >where >the specimen was sent in and how do you dispose the reagents from the >processor and the stainer? >Can this biopsy be processed, once treated with formic acid, along with >the >other specimens or should the biopsy be processed with the other >specimens? > >Need Help!! > >Thank you in advance > >Mala Srishan >Histology Supervisor >Holy Name Hospital >Teaneck, NJ 07666 > > >_______________________________________________ >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 _________________________________________________________________ [1]FREE pop-up blocking with the new MSN Toolbar get it now! References 1. http://g.msn.com/8HMAENUS/2734??PS=47575 From kemlo <@t> tiscali.co.uk Thu May 20 02:13:57 2004 From: kemlo <@t> tiscali.co.uk (Kemlo) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] H and E In-Reply-To: Message-ID: <40A334BB0001F9CD@mk-cpfrontend-3.mail.uk.tiscali.com> A 'flash of periodate' that's like adding sodium monoglutimate to food! I'm no expert, but I prefer the natural ways. Sun ripened haematin, stops that nasty overoxidation and spurious blue staining. The only problem is the lack of the damned Sun in the UK; that' why over the last few days we have flasks everywhere. Organic Gill's, if I may use the term? Kemlo Rogerson MSc MIBiol CBiol CMS DMS CSci FIBMS Chartered Biologist/ Chartered Scientist. __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From marshall <@t> cormack.uct.ac.za Thu May 20 04:46:07 2004 From: marshall <@t> cormack.uct.ac.za (Marshall) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Pas stain(neutral,acid mucin,chief cells Message-ID: <40AC7E5F.C0D8C914@cormack.uct.ac.za> Hi all, A colleague of mine would like to know if the Pas stain stains neutral as well as acid mucins. The acid mucins being sialomucin and sulphated sialomucins. She would also like to know if chief(peptic) cells of the stomach stain with Pas and if so what is the compound which is staining Pas positively. Many Thanks Sharon Marshall Dept. Human Biology University of Cape Town South Africa E-mail: marshall@cormack.uct.ac.za From Andrew.Gray <@t> vcp.monash.edu.au Thu May 20 06:19:46 2004 From: Andrew.Gray <@t> vcp.monash.edu.au (Andrew Gray) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] c-kit Ab: Could someone in Melbourne spare a smidgeon? Message-ID: <130.194.217.87.1085050568.99223@my.monash.edu.au> Dear Histonetters, I'd like to label some ICC's in rat gut (immunofluorescence), to put some icing on the 'cake' that is my thesis (dog's breakfast more like it). I intend to stain a very limited run of sections (<10), so I thought I'd put out a call to see if anyone in Melbourne (Australia) could spare a very small amount of anti-c-kit Ab for this purpose. By means of restitution, I offer the benefit of my extensive experience in IHC problem handling, and kudos in the form of a thesis acknowledgement. If you are interested, please e-mail me. If you have advice about c-kit staining or experience with it on gut nerve plexus whole-mounts, please post a message about it on the Histonet! Best wishes, Andrew Gray PhD student Victorian College of Pharmacy Monash University From Barry.R.Rittman <@t> uth.tmc.edu Thu May 20 07:03:26 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] H and E Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0635A79@UTHEVS3.mail.uthouston.edu> Here is Houston we have summer lasting from May through October. Perhaps a trade of sun for good English sausages would benefit both groups. Barry -----Original Message----- From: Kemlo [mailto:kemlo@tiscali.co.uk] Sent: Thu 5/20/2004 2:13 AM To: Marshall Terry Dr, Consultant Histopathologist; Barry R Rittman; histonet@lists.utsouthwestern.edu Cc: Subject: RE: [Histonet] H and E A 'flash of periodate' that's like adding sodium monoglutimate to food! I'm no expert, but I prefer the natural ways. Sun ripened haematin, stops that nasty overoxidation and spurious blue staining. The only problem is the lack of the damned Sun in the UK; that' why over the last few days we have flasks everywhere. Organic Gill's, if I may use the term? Kemlo Rogerson MSc MIBiol CBiol CMS DMS CSci FIBMS Chartered Biologist/ Chartered Scientist. __________________________________________________ Broadband from an unbeatable ?15.99! http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM From madaryhtl <@t> juno.com Thu May 20 07:40:41 2004 From: madaryhtl <@t> juno.com (madaryhtl@juno.com) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Thanks for letting me back on Histonet! Message-ID: <20040520.054056.28340.32447@webmail12.lax.untd.com> And thanks to all of the folks that helped me get back on. I had to stop for a while(the workplace said get off). So now I use home email. I have to say I missed the banter, and just how much histonet makes you think about your own everyday process. I was "jonesin" for a Jones procedure! I was fishin for a Gill's procedure. Did I just say that? A daily set of checks and balances, and of course stuff that just cracks me up. From ian.montgomery <@t> bio.gla.ac.uk Thu May 20 07:43:44 2004 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:23:00 2005 Subject: Fwd: RE: [Histonet] H and E Message-ID: <6.1.0.6.2.20040520134127.02d12ab0@udcf.gla.ac.uk> Barry, That's a bit cruel, calling the English sausages. Dumplings yes, but sausages, that's taking things a bit far. Ian. >Here is Houston we have summer lasting from May through October. >Perhaps a trade of sun for good English sausages would benefit both groups. >Barry > >-----Original Message----- >From: Kemlo [mailto:kemlo@tiscali.co.uk] >Sent: Thu 5/20/2004 2:13 AM >To: Marshall Terry Dr, Consultant Histopathologist; Barry R Rittman; >histonet@lists.utsouthwestern.edu >Cc: >Subject: RE: [Histonet] H and E > > > > A 'flash of periodate' that's like adding sodium monoglutimate to > food! > > I'm no expert, but I prefer the natural ways. Sun ripened > haematin, stops > that nasty overoxidation and spurious blue staining. The only > problem is > the lack of the damned Sun in the UK; that' why over the last few > days we > have flasks everywhere. Organic Gill's, if I may use the term? > > > Kemlo Rogerson MSc MIBiol CBiol CMS DMS CSci FIBMS > Chartered Biologist/ Chartered Scientist. > > > > __________________________________________________ > Broadband from an unbeatable ??15.99! > > http://www.tiscali.co.uk/products/broadband/home.html?code=SM-NL-11AM > > > > >_______________________________________________ >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 amarusk1 <@t> FAIRVIEW.ORG Thu May 20 07:53:59 2004 From: amarusk1 <@t> FAIRVIEW.ORG (ANN MARUSKA) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] pax 5 Message-ID: Hi Histonetters, Has anyone had luck staining B5 fixed bone marrows with Pax 5? I would be interested to know your secret! 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 pstefanova <@t> sten.sunnybrook.utoronto.ca Thu May 20 08:26:28 2004 From: pstefanova <@t> sten.sunnybrook.utoronto.ca (Petia P Stefanova) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] spleen References: <6.1.0.6.2.20040520134127.02d12ab0@udcf.gla.ac.uk> Message-ID: <001a01c43e6e$0f1537f0$9d194c8e@WS21203> Hi, I have problems to cut mouse spleen. I have already optimized my processing protocol but I still encounter difficulties to get good quality of spleen sections. My cutting angle is good but when I section the tissue comes off the ribbon. I tried to cool it, soak it but it didn't work. My processing protocol is: 10%NBF - 24h 70% ETOH - 45min. 95% ETOH - 45min 95% ETOH - 45min 100%ETOH - 45min 100% ETOH - 45min 100% ETOH - 45min Xylene - 45min Xylene - 45min Wax - 3x1h Any suggestion would be much appreciated Thanks Petia From maxim_71 <@t> mail.ru Thu May 20 08:37:46 2004 From: maxim_71 <@t> mail.ru (=?koi8-r?Q?=22=F0=C5=DB=CB=CF=D7=20=ED=C1=CB=D3=C9=CD=22=20?=) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Fixation and Decalcification of Bone Marrow Bxs. Message-ID: Hi, Charlene We use fixation of specimens in Holland, decal in a equal volumes of 8 % acetic and 8 % hydrochloric acids 1-4 hours. Further - alcohols, clearites and paraffin. We do not use IHC, but H&E, Giemsa, Gram, etc. turn out perfectly. Before staining it is necessary to wash the rests of a fixing agent carefully. We don t see any infringements of morphology. But, we work more often with autopsy specimens. Maxim_71@mail.ru Maxim Peshkov, HT Pathoanatomical bureau, Taganrog, Russia. From srishan <@t> mail.holyname.org Thu May 20 08:49:17 2004 From: srishan <@t> mail.holyname.org (srishan@mail.holyname.org) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] CJD DECONTAMINATION Message-ID: Hello All, Thank you very much to all of you who responded and gave guidance regarding CJD decontamination. I do really appreciate it. Mala Srishan Histology Supervisor Holy Name Hospital Teaneck, NJ 07666 201 833 3023 From deewolfe <@t> anatechltdusa.com Thu May 20 09:04:17 2004 From: deewolfe <@t> anatechltdusa.com (Dee Wolfe) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Alternative fixative users (Prefer) In-Reply-To: Message-ID: <9562B0DE-AA66-11D8-97BB-000A957A311C@anatechltdusa.com> Mary, There is a reference lab that specializes in Prefer fixed tissue. Please contact Dr. Alex Cudkowicz of Cedar Diagnostic at 970.564.2083 or 970.564.2089 (Dr. Cudkowicz's direct line). He can supply you with the growing list of antibodies they have worked up with Prefer. Your pathologist might be interested in speaking with Dr. Cudkowski regarding the validation data supporting the use of Prefer and IHC. Dee Wolfe, Technical Consultant ANATECH LTD. ph: 1.800.262.8324 fax: 1.269.964.8084 deewolfe@anatechltdusa.com www.anatechltdusa.com On Wednesday, May 19, 2004, at 11:22 AM, Mary Bryhan wrote: > Hi, after many years as a Prefer fixative user our pathologists are > telling us that we have to go back to formalin. This came about after > a letter to the editor appeared in the Journal of Surgical Pathology. > > Has anyone out there been approached by their pathologists to switch > back? > > Mary Bryhan > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From rschoon <@t> email.unc.edu Thu May 20 09:19:05 2004 From: rschoon <@t> email.unc.edu (Robert Schoonhoven) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] BDNF IHC References: Message-ID: <40ACBE59.2080709@email.unc.edu> Fellow 'Netter's, I've been having some problems with anti-human BDNF IHC. The clone is MAB248 from R&D Systems. The problem: no staining. Solutions tried: Different titrations, incubation time and temp. as well as HEIR (also tried calling their tech service -nada) Question: Who is doing this procedure and what are you doing that makes it work? What antibody are you suggesting (use). TIA Robert Schoonhoven, Center for Environmental Health and Susceptibility & Lab. of Molecular Carcinogenesis and Mutagenesis UNC-CH 919.966.6343 From rfail <@t> toolkitmail.com Thu May 20 11:05:06 2004 From: rfail <@t> toolkitmail.com (rfail) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] John Dennis Message-ID: <40acd732.39.320a.1842250432@toolkitmail.com> I'm trying to reach Jon Dennis re. C4d Thanks Rena Fail From georgecole <@t> ev1.net Thu May 20 11:15:25 2004 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Tower Of Babel erected on Hematoxylin Message-ID: <000001c43e85$a96900e0$0a4dbad0@hppav> Histotechs; There is everything from smooth adoption and irate rejection of the new means of using Hematoxylin, that barky stuff Ehrilch discovered to be useful in staining tissues long ago. I usually tell about GILLS III, then duck the diatribes that come usually bark back at me. Folks, I mixed my Hematoxylin and ripened it on the roof like the pioneers did. As I more likely got rain on it than maturing sun light here in Oregon, I was delighted to hear about instant mercuric ripening of the stain. I Can't believe it, we used to play with mercury like children. So when hematoxylin preps came along that were already ripened, I thought the glory days were here. Well, all the mixes later---of which Mayers and Harris were dominant for so long, and have lately given way to GILLS I, II, and III mixtures. Heated responses result from discussions of which of these very good mixes, all of them, ranging all the way from you techs who still ripen the stuff on the roof, to those who idolize the GILLs III. Tempers flare. The Tower Of Babel was sweet talking Nice Town compared to some of the heat a tech will take when he tells the world about how well his favorite Hematoxylin stain works. Folks, when ever one of the improvements came along, I got some. I set up my hematoxylin as usual and set up the new candidate and ran them side by side. I then asked all hands---techs, doctors, my wife----which one they liked better. Which ever one won, I kept ( ssshhh, provided I liked it too). The use of GILLS III at my retirement was a result of tests in my lab with residents and pathologists, as well as massive trials in the main lab, comparisons of the old and the new----side-by-side trials on the same tissues---no one cussed---no one drew back---the matter was so straight forward, it was quickly settled. And in every side-by-side trial the GILLS II, at first, and then the GILLS III, won. There was no question about it. The slides did the talking. The rangling on the histonet about Hematoxylins resemble the Scholastic Age of Reverence to Ariistotle----kneeling to Authority---the authority usually being a person's personal habit. Folks, it doesn't really hurt to allow improvements into one's life. The increased quality and the usually simpler processes, are gifts available to the receptive tech. And in the background, the patients being served with the stain investigations are better served. Now then, I sign my name---Gus Swanser.----oh phooey, I might as well put my real name down and duck. georgecole@ev1.net From marytedo <@t> hotmail.com Thu May 20 11:46:57 2004 From: marytedo <@t> hotmail.com (María Teresa Domínguez) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Unsuscribe for vacations Message-ID: I will be out of place from May the 30th, to July the 13th. Ht. Maria Teresa Dominguez Anatomic Pathology Service Hospital Regional Río Grande, Río Grande, Tierra del Fuego, Argentina. _________________________________________________________________ Add photos to your messages with [1]MSN 8. Get 2 months FREE*. References 1. http://g.msn.com/8HMAEN/2749??PS=47575 From Robert.Lott <@t> bhsala.com Thu May 20 11:46:01 2004 From: Robert.Lott <@t> bhsala.com (Lott, Robert) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Info. on FGF-23 Message-ID: <35B6C610DD1DD311B1FA0008C791400407F300D5@gobexchm3.bhsala.com> Subject: Info. on FGF-23 Hello Everyone, Is anyone doing standard Immunohistochemistry with Fibroblast Growth Factor-23 on formalin-fixed paraffin embedded tissues? I found a goat antibody to FGF-23 from Santa Cruz Biotech... are there others that you know about? THANKS!! Robert Robert L. Lott, HTL(ASCP) Manager, Anatomic Pathology Baptist Health System 800 Montclair Road Birmingham, AL 35213 205-592-5388 phone 205-592-5646 fax robert.lott@bhsala.com Confidentiality Notice: The information contained in this email message is privileged and confidential information and intended only for the use of the individual or entity named in the address. If you are not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this information is strictly prohibited. If you received this information in error, please notify the sender and delete this information from your computer and retain no copies of any of this information. From marirose.satterfield <@t> MercyMemorial.org Thu May 20 11:49:48 2004 From: marirose.satterfield <@t> MercyMemorial.org (Satterfield, Marirose) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Cap inspection Message-ID: Our hospital is having their CAP inspection in June. Upon going through our checklist, I feel there are two question we do not have covered. We are a small hospital and only do 1-2 autopsies a year. Could anyone let me know what their facility does to meet the criteria for these two questions? ANP.30100 Are the findings of the postmortem examination used for correlative clinicopathological teaching purposes designed to enhance the quality of patient care? ANP.30575 Are autopsy findings that were clinically inapparent specifically documented and communicated interdepartmentally? Thanks for any help. Mari Satterfield Mercy Memorial Hospital The above signature block is intended for identification purposes only and does not constitute affirmation that a binding contract has been created via this email communication unless expressly so stated within the body of the email message. Further, this message is intended for the use of the individual or entity to which it is addressed and may contain information that is privileged, confidential and/or exempt by law from disclosure. If the reader of this message is not the intended recipient, any further dissemination, distribution or copying of the communication is strictly prohibited. If you have received this communication in error, please notify me immediately. Thank you for your anticipated cooperation. From eshields <@t> bhset.org Thu May 20 12:27:52 2004 From: eshields <@t> bhset.org (E Sharon Shields) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] EGRF Message-ID: Histonetters, Im in the validation process for EGFR. I have noticed that the normal liver cells in liver core biopsies have a positive membrane stain, 2+ - 3+ . The metastic tumor cells are not difficult to identify, since their membrane staining when positive is not nearly as uniform or as intense as the normal liver cells. Has anyone else seen this? No other normal cells in the tissues we have stained stains positive. We are using Ventana antibody on the Benchmark and following the recommend protocol. Sharon Baptist Hospital of E TN Knoxville, TN 865 549-4351 From garygill <@t> dcla.com Thu May 20 11:57:16 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Tower Of Babel erected on Hematoxylin Message-ID: An E.C. Cole published Studies on Hematoxylin Stains in Stain Technology in 1943. Truth be told, hematoxylin formulations are more alike than different with a few notable exceptions. Differences in performance are more often the result of how they're used than what they're made of. In theory, at least, all hematoxylins should produce equivalent visual outcomes when used properly. The composition of some (e.g., Gill's) contribute more to successful outcomes than the composition of most (e.g., Mayer's [contains controlled chemical chloral hydrate {the stuff of "Mickey Finns", aka "knockout drops"}, too much sodium iodate, therefore short useful life {not sold commercially}]), Harris's [too strong, overstains, four variants {full-strength, half-strength, with and without acetic acid}, composition of differentiator {how much HCl dissolved in either water or alcohol?}]). Gary Gill -----Original Message----- From: George Cole [mailto:georgecole@ev1.net] Sent: Thursday, May 20, 2004 11:15 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tower Of Babel erected on Hematoxylin Histotechs; There is everything from smooth adoption and irate rejection of the new means of using Hematoxylin, that barky stuff Ehrilch discovered to be useful in staining tissues long ago. I usually tell about GILLS III, then duck the diatribes that come usually bark back at me. Folks, I mixed my Hematoxylin and ripened it on the roof like the pioneers did. As I more likely got rain on it than maturing sun light here in Oregon, I was delighted to hear about instant mercuric ripening of the stain. I Can't believe it, we used to play with mercury like children. So when hematoxylin preps came along that were already ripened, I thought the glory days were here. Well, all the mixes later---of which Mayers and Harris were dominant for so long, and have lately given way to GILLS I, II, and III mixtures. Heated responses result from discussions of which of these very good mixes, all of them, ranging all the way from you techs who still ripen the stuff on the roof, to those who idolize the GILLs III. Tempers flare. The Tower Of Babel was sweet talking Nice Town compared to some of the heat a tech will take when he tells the world about how well his favorite Hematoxylin stain works. Folks, when ever one of the improvements came along, I got some. I set up my hematoxylin as usual and set up the new candidate and ran them side by side. I then asked all hands---techs, doctors, my wife----which one they liked better. Which ever one won, I kept ( ssshhh, provided I liked it too). The use of GILLS III at my retirement was a result of tests in my lab with residents and pathologists, as well as massive trials in the main lab, comparisons of the old and the new----side-by-side trials on the same tissues---no one cussed---no one drew back---the matter was so straight forward, it was quickly settled. And in every side-by-side trial the GILLS II, at first, and then the GILLS III, won. There was no question about it. The slides did the talking. The rangling on the histonet about Hematoxylins resemble the Scholastic Age of Reverence to Ariistotle----kneeling to Authority---the authority usually being a person's personal habit. Folks, it doesn't really hurt to allow improvements into one's life. The increased quality and the usually simpler processes, are gifts available to the receptive tech. And in the background, the patients being served with the stain investigations are better served. Now then, I sign my name---Gus Swanser.----oh phooey, I might as well put my real name down and duck. georgecole@ev1.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Thu May 20 12:52:31 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Tower Of Babel erected on Hematoxylin Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA206EB2C@usca0082k08.labvision.apogent.com> Geroge cole wrote: << Folks, I mixed my Hematoxylin and ripened it on the roof like the pioneers did. As I more likely got rain on it than maturing sun light here in Oregon,...>> Geroge, is it true that Oregonians rust rather than tan...? Tim Morken (in sunny california) -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Thursday, May 20, 2004 9:57 AM To: 'George Cole'; 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] Tower Of Babel erected on Hematoxylin An E.C. Cole published Studies on Hematoxylin Stains in Stain Technology in 1943. Truth be told, hematoxylin formulations are more alike than different with a few notable exceptions. Differences in performance are more often the result of how they're used than what they're made of. In theory, at least, all hematoxylins should produce equivalent visual outcomes when used properly. The composition of some (e.g., Gill's) contribute more to successful outcomes than the composition of most (e.g., Mayer's [contains controlled chemical chloral hydrate {the stuff of "Mickey Finns", aka "knockout drops"}, too much sodium iodate, therefore short useful life {not sold commercially}]), Harris's [too strong, overstains, four variants {full-strength, half-strength, with and without acetic acid}, composition of differentiator {how much HCl dissolved in either water or alcohol?}]). Gary Gill -----Original Message----- From: George Cole [mailto:georgecole@ev1.net] Sent: Thursday, May 20, 2004 11:15 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tower Of Babel erected on Hematoxylin Histotechs; There is everything from smooth adoption and irate rejection of the new means of using Hematoxylin, that barky stuff Ehrilch discovered to be useful in staining tissues long ago. I usually tell about GILLS III, then duck the diatribes that come usually bark back at me. Folks, I mixed my Hematoxylin and ripened it on the roof like the pioneers did. As I more likely got rain on it than maturing sun light here in Oregon, I was delighted to hear about instant mercuric ripening of the stain. I Can't believe it, we used to play with mercury like children. So when hematoxylin preps came along that were already ripened, I thought the glory days were here. Well, all the mixes later---of which Mayers and Harris were dominant for so long, and have lately given way to GILLS I, II, and III mixtures. Heated responses result from discussions of which of these very good mixes, all of them, ranging all the way from you techs who still ripen the stuff on the roof, to those who idolize the GILLs III. Tempers flare. The Tower Of Babel was sweet talking Nice Town compared to some of the heat a tech will take when he tells the world about how well his favorite Hematoxylin stain works. Folks, when ever one of the improvements came along, I got some. I set up my hematoxylin as usual and set up the new candidate and ran them side by side. I then asked all hands---techs, doctors, my wife----which one they liked better. Which ever one won, I kept ( ssshhh, provided I liked it too). The use of GILLS III at my retirement was a result of tests in my lab with residents and pathologists, as well as massive trials in the main lab, comparisons of the old and the new----side-by-side trials on the same tissues---no one cussed---no one drew back---the matter was so straight forward, it was quickly settled. And in every side-by-side trial the GILLS II, at first, and then the GILLS III, won. There was no question about it. The slides did the talking. The rangling on the histonet about Hematoxylins resemble the Scholastic Age of Reverence to Ariistotle----kneeling to Authority---the authority usually being a person's personal habit. Folks, it doesn't really hurt to allow improvements into one's life. The increased quality and the usually simpler processes, are gifts available to the receptive tech. And in the background, the patients being served with the stain investigations are better served. Now then, I sign my name---Gus Swanser.----oh phooey, I might as well put my real name down and duck. georgecole@ev1.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From emry <@t> u.washington.edu Thu May 20 12:56:01 2004 From: emry <@t> u.washington.edu (P. Emry) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Tower Of Babel erected on Hematoxylin In-Reply-To: <000001c43e85$a96900e0$0a4dbad0@hppav> References: <000001c43e85$a96900e0$0a4dbad0@hppav> Message-ID: Oratory....ah....interested in running for office? Oh, Please? Groovy, George Cole, .....Gus Swanzer...there has to be a story there... Trisha Seattle On Thu, 20 May 2004, George Cole wrote: > Histotechs; > There is everything from smooth adoption and irate rejection of the new > means of using Hematoxylin, that barky stuff Ehrilch discovered to be > useful in staining tissues long ago. I usually tell about GILLS III, > then duck the diatribes that come usually bark back at me. Folks, I > mixed my Hematoxylin and ripened it on the roof like the pioneers did. > As I more likely got rain on it than maturing sun light here in Oregon, > I was delighted to hear about instant mercuric ripening of the stain. I > Can't believe it, we used to play with mercury like children. So when > hematoxylin preps came along that were already ripened, I thought the > glory days were here. Well, all the mixes later---of which Mayers and > Harris were dominant for so long, and have lately given way to GILLS I, > II, and III mixtures. Heated responses result from discussions of which > of these very good mixes, all of them, ranging all the way from you > techs who still ripen the stuff on the roof, to those who idolize the > GILLs III. Tempers flare. The Tower Of Babel was sweet talking Nice > Town compared to some of the heat a tech will take when he tells the > world about how well his favorite Hematoxylin stain works. Folks, when > ever one of the improvements came along, I got some. I set up my > hematoxylin as usual and set up the new candidate and ran them side by > side. I then asked all hands---techs, doctors, my wife----which one > they liked better. Which ever one won, I kept ( ssshhh, provided I > liked it too). The use of GILLS III at my retirement was a result of > tests in my lab with residents and pathologists, as well as massive > trials in the main lab, comparisons of the old and the > new----side-by-side trials on the same tissues---no one cussed---no one > drew back---the matter was so straight forward, it was quickly settled. > And in every side-by-side trial the GILLS II, at first, and then the > GILLS III, won. There was no question about it. The slides did the > talking. The rangling on the histonet about Hematoxylins resemble the > Scholastic Age of Reverence to Ariistotle----kneeling to > Authority---the authority usually being a person's personal habit. > Folks, it doesn't really hurt to allow improvements into one's life. The > increased quality and the usually simpler processes, are gifts available > to the receptive tech. And in the background, the patients being served > with the stain investigations are better served. > Now then, I sign my name---Gus Swanser.----oh phooey, I might as well > put my real name down and duck. > georgecole@ev1.net > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From la.sebree <@t> hosp.wisc.edu Thu May 20 13:13:50 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] EGRF Message-ID: Are you using AB block, Sharon? You may need to block biotin off line also since VMS' AB block is not real strong. 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 -----Original Message----- From: E Sharon Shields [mailto:eshields@bhset.org] Sent: Thursday, May 20, 2004 12:28 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] EGRF Histonetters, Im in the validation process for EGFR. I have noticed that the normal liver cells in liver core biopsies have a positive membrane stain, 2+ - 3+ . The metastic tumor cells are not difficult to identify, since their membrane staining when positive is not nearly as uniform or as intense as the normal liver cells. Has anyone else seen this? No other normal cells in the tissues we have stained stains positive. We are using Ventana antibody on the Benchmark and following the recommend protocol. Sharon Baptist Hospital of E TN Knoxville, TN 865 549-4351 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ploykasek <@t> phenopath.com Thu May 20 13:06:18 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] EGRF In-Reply-To: Message-ID: Sharon - we have seen this staining in liver (we are using a different clone of EGFR). You can see staining in normal colon & on nerves, too. Patti Loykasek > Histonetters, > Im in the validation process for EGFR. I have noticed that the normal liver > cells in liver core biopsies have a positive membrane stain, 2+ - 3+ . The > metastic tumor cells are not difficult to identify, since their membrane > staining when positive is not nearly as uniform or as intense as the normal > liver cells. Has anyone else seen this? No other normal cells in the tissues > we have stained stains positive. We are using Ventana antibody on the > Benchmark and following the recommend protocol. > > Sharon > Baptist Hospital of E TN > Knoxville, TN > 865 549-4351 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From awall <@t> unmc.edu Thu May 20 13:19:11 2004 From: awall <@t> unmc.edu (awall@unmc.edu) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] (no subject) Message-ID: Hello People, This is my first posting. I am currently in the process of staining bone and cartilage of newly born mice. I am using Alizarin Red S for the bones and Alcian Blue for the cartilage. I eviscerate as much of the skin and muscle tissue as I can. The problem is I am not getting the cartilage and the bone to stain correctly. I received these samples from another lab and found out they had been fixed and stored @ 4 degrees in the same paraformaldehyde for more than a year. The bone stains bluish green and the cartilage does not stain at all. Any suggestion would be a big help and I have plenty of specimens to work with so I will take what I can get. Thank you for your time, Drew University of Nebraska Medical Center From rfail <@t> toolkitmail.com Thu May 20 13:32:44 2004 From: rfail <@t> toolkitmail.com (rfail) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] OCT/IHC Message-ID: <40acf9cc.392.2516.1595231356@toolkitmail.com> Is there a way to completely remove OCT before staining for immunohistochemistry? Thanks Rena Fail HT(ASCP)QIHC Medical university of South Carolina Charleston, SC From lori.karnes <@t> saskatoonhealthregion.ca Thu May 20 13:40:22 2004 From: lori.karnes <@t> saskatoonhealthregion.ca (Karnes, Lori SktnHR) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] M. leprae controls Message-ID: <6841F8237DF2D711A644009027E8728F67818D@sdhmail.saskatoonhealthregion.ca> I was wondering if anyone knows where I can purchase M. Leprae controls for our Fite stain. I checked with Histonet archives and there was info about staining but not about where to purchase control slides. Thank you in advance. Lori Karnes From eca9 <@t> georgetown.edu Thu May 20 14:18:26 2004 From: eca9 <@t> georgetown.edu (Eva C. Andersson) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Formalin fixation of cells for paraffin embedding? Message-ID: <40AD0482.4010606@georgetown.edu> Hi everyone, My lab wants to paraffin embed some cells to be used as positive controls for immunohisochemistry staining. We have a cell line growing but need a good protocol for fixing the cells with formalin. Once the cells have been fixed the pellet will be embedded in paraffin by our histopathology department. Does anyone have a good and easy method for the fixation part? We will deliver the pellet to the histopathology department in 70% ethanol. Thanks in advance, Eva Andersson Research Assistant Georgetown University From funderwood <@t> mcohio.org Thu May 20 14:19:18 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] M. leprae controls Message-ID: American MasterTech Scientific carreis them. 1.800.860.4073 www.mastertechs.com >>> "Karnes, Lori SktnHR" 05/20/04 02:40PM >>> I was wondering if anyone knows where I can purchase M. Leprae controls for our Fite stain. I checked with Histonet archives and there was info about staining but not about where to purchase control slides. Thank you in advance. Lori Karnes _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From namorant <@t> seattlecca.org Thu May 20 15:03:47 2004 From: namorant <@t> seattlecca.org (Amoranto, Nelson N) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] acid alcohjol Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B9486C1286@wala01.seattlecca.org> Hello fellow Histotechs, You only need a long dip or two quick dips in acid alcohol to differentiate the excess Hematoxylin..... namoranto,seattle, wa. 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 madaryhtl <@t> juno.com Thu May 20 15:52:03 2004 From: madaryhtl <@t> juno.com (madaryhtl@juno.com) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] spleen microtomy Message-ID: <20040520.135237.11063.40878@webmail03.lax.untd.com> Wash in water after the formalin(are the spleens cut to allow the fix to penetrate past the capsule?), and cut down your times in alcohols to 30 minutes and no heat of course. From David.Muskett <@t> btopenworld.com Thu May 20 16:03:20 2004 From: David.Muskett <@t> btopenworld.com (David Muskett) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Reviewing Decal Procedures Message-ID: Dear Histonetters We are currently reviewing our decal procedures. I am writing to ask what the best routine decalcifying agent is? We are currently using a weak HCl method with some Chloral Hydrate in it. This seems less than ideal as a decalcifying agent and I would like to reduce the use of chloral hydrate in the lab as a safety feature. Hope you can help Regards David Muskett Liverpool, UK From bgoldman <@t> wam.umd.edu Thu May 20 16:54:32 2004 From: bgoldman <@t> wam.umd.edu (Beth A. Goldman) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] pH of cresyl violet Message-ID: Hello. Does anyone know what the optimal pH for cresyl violet stain should be? I am staining brain sections that have been fixed with Notox histo (a formalin-free fixative). Thanks! From garygill <@t> dcla.com Thu May 20 17:09:27 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] acid alcohjol Message-ID: Your recommendation is correct as long as others do use/do what you do: your particular stain, concentration, age, specimen, concentration of acid and alcohol. Therefore, you must provide these details if they are to be useful. Otherwise, you paint with too broad a brush. The devil resides in the details: dye left = (dye in) minus (dye out). Gary Gill -----Original Message----- From: Amoranto, Nelson N [mailto:namorant@seattlecca.org] Sent: Thursday, May 20, 2004 3:04 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] acid alcohjol Hello fellow Histotechs, You only need a long dip or two quick dips in acid alcohol to differentiate the excess Hematoxylin..... namoranto,seattle, wa. 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> colobio.com Thu May 20 18:41:01 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] looking for antibody to LAMP2 Message-ID: Folks, The Cardio people asked me to look for an antibody to a protein called LAMP2 There are 2 isoforms of this protein (alpha and beta) that are basically identical except for the most extreme carboxyl terminus. Anybody know this protein? Patsy From pengbw <@t> sjtu.edu.cn Thu May 20 19:49:13 2004 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] OCT/IHC Message-ID: <20040521004913.1DDD110E1D70@sjtu.edu.cn> Hi, Rena Washing in PBS 5min, 2x can remove OCT and is qualified for most IHC. But what do you mean by completely remove OCT? Baowei Peng Shanghai Jiaotong University Shanghai China ----- Original Message ----- From: rfail To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] OCT/IHC Is there a way to completely remove OCT before staining for immunohistochemistry? Thanks Rena Fail HT(ASCP)QIHC Medical university of South Carolina Charleston, SC _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RFail <@t> Charleston.net Thu May 20 20:14:00 2004 From: RFail <@t> Charleston.net (Rena Fail) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] OCT/IHC rsponse In-Reply-To: <40acf9cc.392.2516.1595231356@toolkitmail.com> Message-ID: <003401c43ed0$e8267290$9911a6a5@rena> Thank all of you who responded, it appears I did not wash long enough Rena -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of rfail Sent: Thursday, May 20, 2004 2:33 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] OCT/IHC Is there a way to completely remove OCT before staining for immunohistochemistry? Thanks Rena Fail HT(ASCP)QIHC Medical university of South Carolina Charleston, SC _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cgfields <@t> lexhealth.org Fri May 21 06:51:35 2004 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Antibody OLIG2 Message-ID: We are hunting an antibody, OLIG2, oligodendroglioma which is for a brain tumor. Our Pathologist says its being used in research but doesn't know if it's available from anyone commercially yet. He says it's the first marker that marks "specifically" for this tumor. If anyone is using this and knows where it can be purchased it would be appreciated. Thank you the help. Posted for : David B. Johnson UCI Medical Center Irvine Orange, California . _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From MAUGER <@t> email.chop.edu Fri May 21 06:51:33 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Formalin fixation of cells for paraffin embedding? Message-ID: To Eva, Wrap the pellet in lens paper,and put it in a cassette to fix and process with your tissues. I would skip the alcohol fixation and spin the cells down in NBF, then proceed with processing and embed as usual. Some antigens don't like alcohol fix. Jo >>> "Eva C. Andersson" 05/20/04 03:18PM >>> Hi everyone, My lab wants to paraffin embed some cells to be used as positive controls for immunohisochemistry staining. We have a cell line growing but need a good protocol for fixing the cells with formalin. Once the cells have been fixed the pellet will be embedded in paraffin by our histopathology department. Does anyone have a good and easy method for the fixation part? We will deliver the pellet to the histopathology department in 70% ethanol. Thanks in advance, Eva Andersson Research Assistant Georgetown University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Fri May 21 08:09:30 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] (no subject) Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0635A7D@UTHEVS3.mail.uthouston.edu> Welcome or as Charlotte would have said salutations. The time of fixation in formalin should not be the problem with this staining unless the fixative was or became acidic with possible loss of mineral and other components. The major problem with staining whole embryos is adequate penetration of the stain. This is often slowed down by lipid left in the tissues or insufficient time in the staining solutions. Would you let us know the precise procedure that you used please? Thanks and have a great weekend. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of awall@unmc.edu Sent: Thursday, May 20, 2004 1:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Hello People, This is my first posting. I am currently in the process of staining bone and cartilage of newly born mice. I am using Alizarin Red S for the bones and Alcian Blue for the cartilage. I eviscerate as much of the skin and muscle tissue as I can. The problem is I am not getting the cartilage and the bone to stain correctly. I received these samples from another lab and found out they had been fixed and stored @ 4 degrees in the same paraformaldehyde for more than a year. The bone stains bluish green and the cartilage does not stain at all. Any suggestion would be a big help and I have plenty of specimens to work with so I will take what I can get. Thank you for your time, Drew University of Nebraska Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eca9 <@t> georgetown.edu Fri May 21 08:36:12 2004 From: eca9 <@t> georgetown.edu (Eva C. Andersson) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Histogel? Message-ID: <40AE05CC.9060201@georgetown.edu> Hi everyone, I have a question about how to get a hold of Histogel. I have tried ordering it from Fisher but it is on backorder. Can anyone provide me with a different company that sells it? Thanks, Eva Research Assistant Georgetown University From garygill <@t> dcla.com Fri May 21 08:40:09 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Histogel? Message-ID: Richard-Allan Scientific is the manufacturer. RAS also sells it, of course. -----Original Message----- From: Eva C. Andersson [mailto:eca9@georgetown.edu] Sent: Friday, May 21, 2004 8:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histogel? Hi everyone, I have a question about how to get a hold of Histogel. I have tried ordering it from Fisher but it is on backorder. Can anyone provide me with a different company that sells it? Thanks, Eva Research Assistant Georgetown University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From FreidaC <@t> aol.com Fri May 21 08:46:13 2004 From: FreidaC <@t> aol.com (FreidaC@aol.com) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Embedding Message-ID: <6d.2a2669fa.2ddf6225@aol.com> I have a question for all of the derm and hard tissue people out there. How do you embed the sections? Do you cut the dense or hard tissue first or last - or do you embed at an angle? One individual is questioning the answer given in the study guide and so far I have different answers from those I have consulted. Thanks, Freida Carson From marshal <@t> dcla.com Fri May 21 08:52:11 2004 From: marshal <@t> dcla.com (Marsha Linville) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Formalin fixation of cells for paraffin embedding? Message-ID: Joanne, Would one of the antigens that you speak of that doesn't like alcohol fixation be Thyroglobulin? Marsha Linville DCL Medical Laboratory Indianapolis, IN 46268 (317) 874-1232 -----Original Message----- From: Joanne Mauger [mailto:MAUGER@email.chop.edu] Sent: Friday, May 21, 2004 6:52 AM To: eca9@georgetown.edu; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Formalin fixation of cells for paraffin embedding? To Eva, Wrap the pellet in lens paper,and put it in a cassette to fix and process with your tissues. I would skip the alcohol fixation and spin the cells down in NBF, then proceed with processing and embed as usual. Some antigens don't like alcohol fix. Jo >>> "Eva C. Andersson" 05/20/04 03:18PM >>> Hi everyone, My lab wants to paraffin embed some cells to be used as positive controls for immunohisochemistry staining. We have a cell line growing but need a good protocol for fixing the cells with formalin. Once the cells have been fixed the pellet will be embedded in paraffin by our histopathology department. Does anyone have a good and easy method for the fixation part? We will deliver the pellet to the histopathology department in 70% ethanol. Thanks in advance, Eva Andersson Research Assistant Georgetown 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 MAUGER <@t> email.chop.edu Fri May 21 09:10:58 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Formalin fixation of cells for paraffinembedding? Message-ID: Marsha, I'm not sure about thyroglobulin, but I know S-100 positivety diminishes or disappears after alcohol fixation. If its well fixed in NBF first, the following alcs don't seem to hurt. Jo >>> "Marsha Linville" 05/21/04 09:52AM >>> Joanne, Would one of the antigens that you speak of that doesn't like alcohol fixation be Thyroglobulin? Marsha Linville DCL Medical Laboratory Indianapolis, IN 46268 (317) 874-1232 -----Original Message----- From: Joanne Mauger [mailto:MAUGER@email.chop.edu] Sent: Friday, May 21, 2004 6:52 AM To: eca9@georgetown.edu; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Formalin fixation of cells for paraffin embedding? To Eva, Wrap the pellet in lens paper,and put it in a cassette to fix and process with your tissues. I would skip the alcohol fixation and spin the cells down in NBF, then proceed with processing and embed as usual. Some antigens don't like alcohol fix. Jo >>> "Eva C. Andersson" 05/20/04 03:18PM >>> Hi everyone, My lab wants to paraffin embed some cells to be used as positive controls for immunohisochemistry staining. We have a cell line growing but need a good protocol for fixing the cells with formalin. Once the cells have been fixed the pellet will be embedded in paraffin by our histopathology department. Does anyone have a good and easy method for the fixation part? We will deliver the pellet to the histopathology department in 70% ethanol. Thanks in advance, Eva Andersson Research Assistant Georgetown 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Fri May 21 09:11:12 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Embedding Message-ID: I worked in a derm lab for a few years, and it seemed best to embed the skins on a slight angle to the mold, so that when you cut through the soft tissue first, your knife wasn't hitting a straight line of (sometimes) hard epidermis. Also, if you cut through the epidermis first, there is a tendency to damage the knife edge, or drag hard particles through the soft tissue, creating scratch artifacts in your section. Jackie O' FreidaC@aol.com Sent by: histonet-bounces@lists.utsouthwestern.edu 05/21/2004 08:46 AM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Embedding I have a question for all of the derm and hard tissue people out there. How do you embed the sections? Do you cut the dense or hard tissue first or last - or do you embed at an angle? One individual is questioning the answer given in the study guide and so far I have different answers from those I have consulted. Thanks, Freida Carson _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From clarkda <@t> ohsu.edu Fri May 21 09:13:33 2004 From: clarkda <@t> ohsu.edu (David Clark) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Embedding Message-ID: Frieda, I was trained to always cut the less dense tissue first. In doing Mohs I always cut thru the underly dermis before the epidermis. I also try to embed the tissue at a 45 degree angle with the epidermis on the right. In my experience there is a great deal of difficulty in trying to cut thru the epidermis first. With bone or hard tissue I also try to cut thru the less dense first, if at all possible. David Clark Kaiser NW & OHSU >>> 05/21/04 6:46 AM >>> I have a question for all of the derm and hard tissue people out there. How do you embed the sections? Do you cut the dense or hard tissue first or last - or do you embed at an angle? One individual is questioning the answer given in the study guide and so far I have different answers from those I have consulted. Thanks, Freida Carson _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bill501 <@t> mindspring.com Fri May 21 09:54:57 2004 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Formalin fixation of cells for paraffin embedding? In-Reply-To: References: Message-ID: At 8:52 AM -0500 5/21/04, Marsha Linville wrote: >Wrap the pellet in lens paper,and put it in a cassette to fix and process >with your tissues. We use those papers the hairdressers use when curling hair. Much cheaper than lens paper. Bill -- ______________ Bill Blank, MD Heartland Lab, Inc From Sue.Kapoor <@t> uhsi.org Fri May 21 10:24:39 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Mesenteric node with obvious tumor Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F55EB@khmcexch.uhsi.org> To any experts that can help, I'm posting this from my pathologist: "We have biopsies of mesenteric node with obvious tumor, morphologically compatible with a neuroendocrine tumor from pancreas, (patient has a mass in the tail of the pancreas). Patient also has history of transitional cell of bladder, which would not be a good histologic fit with the metastatic in the node. CK7, CK20 and CEA are negative as is Chromogranin. I know CK7 and CK20 negativity is typical for lung small cell, but I can find no reference for pancreatic neuro-endocrine tumors. The negative Chromogranin is also puzzling, any ideas??" Thanks much, Dr. James Kranz please reply to: sue.kapoor@uhsi.org THANK YOU! From T.J.A.vanEijl <@t> pharm.uu.nl Fri May 21 11:28:39 2004 From: T.J.A.vanEijl <@t> pharm.uu.nl (Sven van Eijl) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Dead-End system & BMA/MMA embedded tissue Message-ID: <3.0.6.32.20040521172839.0087fd60@solis-mail.uu.nl> Dear Histonetters, This will be my first posting to this list, and I am by no means a real histologists, so bear with me please... We are having a problem using Promega's Dead-End colorimetric TUNEL kit on plastic embedded lungs. In the manual are only guidelines for formalin fixed and parrafin embedded or frozen tissue. We fixed the lungs using Carnoy's and embedded in BMA/MMA. I have had some tips from Promega about modifications to the protocol to account for the use of other fixatives than formalin, but am wondering if it is the fixative or the plastic that is causing the problems. We get a nice brown staining of all nuclei at all times, also in the negative controls. It is not a question of general high background, the rest of the tissue is clear. Any suggestions are highly appreciated. Thanks, Sven **************************************************************************************************************** T.J.A.van Eijl, MSc. Dept. Pharmacology and Pathophysiology Utrecht Institute for Pharmaceutical Sciences Utrecht University Utrecht The Netherlands F.A.F.C. Wentgebouw Sorbonnelaan 16 3584 CA Utrecht Room W146 Tel: +31(0)30 253 1215 Fax: +31(0)30 253 7420 **************************************************************************************************************** "Many a man fails to become a thinker for the sole reason that his memory is too good" Nietzsche **************************************************************************************************************** From Barry.R.Rittman <@t> uth.tmc.edu Fri May 21 10:39:24 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Embedding Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0635A7E@UTHEVS3.mail.uthouston.edu> Freida I totally agree with Jackie especially when talking about paraffin sections. As most skin has epidermal rete pegs it is not possible to cut from the ideal direction, i.e. not encountering the epidermis until after you have passed through all the dermis. I used to cut dermis side first but always at an angle. If you need to set your pathologists mind at rest you can refer him/her to Steedmans book where the mechanics of cutting are detailed. Steedman H.F. 1960. Section Cutting in Microscopy. Blackwell scientific Publications, Oxford, England. There is also useful information to be obtained, even though it is specifically for electron microscopy, in "Thin Sectioning and associated techniques for electron microscopy. Originally put out by Sorvall. Third edition is 1974. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jackie.O'Connor@abbott.com Sent: Friday, May 21, 2004 9:11 AM To: FreidaC@aol.com Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Embedding I worked in a derm lab for a few years, and it seemed best to embed the skins on a slight angle to the mold, so that when you cut through the soft tissue first, your knife wasn't hitting a straight line of (sometimes) hard epidermis. Also, if you cut through the epidermis first, there is a tendency to damage the knife edge, or drag hard particles through the soft tissue, creating scratch artifacts in your section. Jackie O' FreidaC@aol.com Sent by: histonet-bounces@lists.utsouthwestern.edu 05/21/2004 08:46 AM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Embedding I have a question for all of the derm and hard tissue people out there. How do you embed the sections? Do you cut the dense or hard tissue first or last - or do you embed at an angle? One individual is questioning the answer given in the study guide and so far I have different answers from those I have consulted. Thanks, Freida Carson _______________________________________________ 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 BRobert <@t> ameripath.com Fri May 21 11:08:04 2004 From: BRobert <@t> ameripath.com (BRobert@ameripath.com) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] slide disposal Message-ID: How do you guys dispose of your histo slides after 10 years? We are having them hauled away as biohazardous material and it seems pretty expensive to me to do it this way but it seems that the main reason for doing it is for confidentiality purposes as labels have the patient's names on. Anybody knows of a better way (less expensive) to do it? Brigitte Robert Histology Laboratory Manager APMG Inc. 408-399-5050 ext 106 fax: 408-354-4581 From Charlene.Henry <@t> STJUDE.ORG Fri May 21 11:08:32 2004 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Formalin fixation of cells for paraffin embedding? Message-ID: <5CB39BCA5724F349BCB748675C6CA1A2315838@SJMEMXMB02.stjude.sjcrh.local> We process a lot of cell lines for research and have found that if you intend to perform IHC or ISH on the FFPE cells, the following protocol works great. The PBS washes the cells of excess medium used to grow the cells and yields clean IHC. The Hist Gel yields a cell pellet with an even distribution of cells through-out without the cells being clumped together. 1. Suspend cells in 10% NFB to fix cells. 2. Spend down suspended cells and draw off 10% NBF. 3. Wash cells in PBS by adding 2-3 ml of PBS and re-suspend cells with vortex. 4. Spend down cells and draw off PBS. 5. Wash cells in PBS one more time as in step 3 & 4. 6. Add volume of Histo Gel equal to the amount of cells and re-suspend cells with vortex. 7. Allow gel time to become firm (usually 5-10 minutes. 8. Remove gel pellet carefully, cut in half, place in lens paper, and process. Hope this helps because we tried multiple protocols before this one. Thanks, Charlene Henry, HT QIHC Histology/IHC Section Head Department of Pathology St. Jude Children's Research Hospital 332 North Lauderdale Memphis, TN 38105 Phone: 901-495-3349 FAX: 901-495-3100 -----Original Message----- From: Bill Blank [mailto:bill501@mindspring.com] Sent: Friday, May 21, 2004 9:55 AM To: histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Formalin fixation of cells for paraffin embedding? At 8:52 AM -0500 5/21/04, Marsha Linville wrote: >Wrap the pellet in lens paper,and put it in a cassette to fix and process >with your tissues. We use those papers the hairdressers use when curling hair. Much cheaper than lens paper. Bill -- ______________ Bill Blank, MD Heartland Lab, Inc _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From John.Auld <@t> whnt.nhs.uk Fri May 21 11:26:47 2004 From: John.Auld <@t> whnt.nhs.uk (John Auld) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] Decal Message-ID: Dave Oh MY GOD you're still using choral hydrate - stop this immediately!!! 10% formic acid is IMO the best for bone 5% for BMTs. You will get good immuno after this as well, provided you fix properly first. By the way if you use Mayer's haematoxylin the chloral hydrate can be omitted from that as well. It allegedly acts as a preservative and has no effect on staining. Thanks for sending the booklets John From Sue.Kapoor <@t> uhsi.org Fri May 21 11:43:22 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] test Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F55EC@khmcexch.uhsi.org> From madaryhtl <@t> juno.com Fri May 21 12:01:03 2004 From: madaryhtl <@t> juno.com (madaryhtl@juno.com) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] bone and cart staining in the mouse tissue Message-ID: <20040521.100144.24094.51559@webmail20.lax.untd.com> I would not limiot yourself to those two stains although they are great and easy. Add a VOn Kossa maybe for the calcium, and perhaps a panel of different ph's for the Alcian Blue and also maybe an AB/PAS for fun. I thought I might have read something about one of the older hematxylins years ago staining bone/cartilage. Erlichs, Delafiedls? From jcline <@t> wchsys.org Fri May 21 12:06:33 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:23:00 2005 Subject: [Histonet] slide disposal In-Reply-To: Message-ID: <000501c43f55$f81592f0$1d2a14ac@wchsys.org> Since the tissue is processed and sealed, we just throw them away in our trash compactor. The compactor trash is hauled away to the landfill. (Our slides have no patient name on the label) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of BRobert@ameripath.com Sent: Friday, May 21, 2004 12:08 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] slide disposal How do you guys dispose of your histo slides after 10 years? We are having them hauled away as biohazardous material and it seems pretty expensive to me to do it this way but it seems that the main reason for doing it is for confidentiality purposes as labels have the patient's names on. Anybody knows of a better way (less expensive) to do it? Brigitte Robert Histology Laboratory Manager APMG Inc. 408-399-5050 ext 106 fax: 408-354-4581 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***** 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 madaryhtl <@t> juno.com Fri May 21 12:16:02 2004 From: madaryhtl <@t> juno.com (madaryhtl@juno.com) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] I think Chloral Hydrate works better! Message-ID: <20040521.101613.24094.51760@webmail20.lax.untd.com> I make my own Mayers and have tried several recipes with so-called Chloral Hydrate substitutes and even no Chloral Hyrdate. I have done side-by-side studies(not published, there's an idea), and CHloral Hydrate does make a difference in the short run and in the long run if the stuff sits in the bottle longer than you expect. I say keep the Chloral Hydrate. Nick MAdary, B.S. HT/HTL(ASCP)QIHC Histology Manager Human Genome Sciences Rockville, Md 20850 From Sue.Kapoor <@t> uhsi.org Fri May 21 12:18:55 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] mesenteric node Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F55ED@khmcexch.uhsi.org> To any experts that can help, I'm posting this from my pathologist: "We have biopsies of mesenteric node with obvious tumor, morphologically compatible with a neuroendocrine tumor from pancreas, (patient has a mass in the tail of the pancreas). Patient also has history of transitional cell of bladder, which would not be a good histologic fit with the metastatic in the node. CK7, CK20 and CEA are negative as is Chromogranin. I know CK7 and CK20 negativity is typical for lung small cell, but I can find no reference for pancreatic neuro-endocrine tumors. The negative Chromogranin is also puzzling, any ideas??" Thanks much, Dr. James Kranz please reply to: sue.kapoor@uhsi.org THANK YOU! From browning <@t> HHSC.CA Fri May 21 12:46:27 2004 From: browning <@t> HHSC.CA (Browning Deb) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] BCL6, EDTA buffer Message-ID: <3AADFB88753AD31189C100902786B91C0E2781EC@hch_nt_exchange.hhsc.ca> Good afternoon: As much as I dislike EDTA buffer due to the poor morphology (especially if not properly fixed) that it renders, I have not had much success with BCL6 staining with any other buffers. I would appreciate any information on how to improve the morphology of the tissue, the use of other buffers, or any success stories, since I just can't seem to pull the rabbit out of the hat here. Thank you. Debra Browning, ART Technical Specialist, Immunohistochemistry Anatomic Pathology Hamilton Health Sciences phone: (905) 527-4322 ext 46131 e-mail: browning@hhsc.ca fax: (905) 524-2681 This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From pruegg <@t> colobio.com Fri May 21 13:33:46 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Histogel make your own In-Reply-To: Message-ID: A while back someone gave us the recipe for making up histogel type media and I misplaced the message, if you are out there could you send it to me again, thank you, Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gary Gill Sent: Friday, May 21, 2004 7:40 AM To: 'Eva C. Andersson'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histogel? Richard-Allan Scientific is the manufacturer. RAS also sells it, of course. -----Original Message----- From: Eva C. Andersson [mailto:eca9@georgetown.edu] Sent: Friday, May 21, 2004 8:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histogel? Hi everyone, I have a question about how to get a hold of Histogel. I have tried ordering it from Fisher but it is on backorder. Can anyone provide me with a different company that sells it? Thanks, Eva Research Assistant Georgetown 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 dmnelson <@t> iastate.edu Fri May 21 14:16:10 2004 From: dmnelson <@t> iastate.edu (Diane M Nelson) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Decals ( att: David ) Message-ID: <6.1.0.6.2.20040521140936.01b52e68@dmnelson.mail.iastate.edu> Hi David, We use 25% formic acid. For small tissues it generally takes a couple days, for large tissue, five to seven days. We chemically check our decals to see when they are ready. Hope this helps you. Diane From japoteete <@t> saintfrancis.com Fri May 21 14:18:07 2004 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] BCL6, EDTA buffer Message-ID: We use DAKO's BCL6 with DAKO High pH buffer, 30 minutes in a steamer, with no problems. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital Tulsa, OK japoteete@saintfrancis.com > -----Original Message----- > From: Browning Deb [SMTP:browning@HHSC.CA] > Sent: Friday, May 21, 2004 12:46 PM > To: 'histonet@lists.utsouthwestern.edu' > Subject: [Histonet] BCL6, EDTA buffer > > Good afternoon: As much as I dislike EDTA buffer due to the poor > morphology > (especially if not properly fixed) that it renders, I have not had much > success with BCL6 staining with any other buffers. I would appreciate any > information on how to improve the morphology of the tissue, the use of > other > buffers, or any success stories, since I just can't seem to pull the > rabbit > out of the hat here. Thank you. > > Debra Browning, ART > Technical Specialist, Immunohistochemistry > Anatomic Pathology > Hamilton Health Sciences > phone: (905) 527-4322 ext 46131 > e-mail: browning@hhsc.ca > fax: (905) 524-2681 > > > This information is directed in confidence solely to the person named > above > and may not otherwise be distributed, copied or disclosed. Therefore, > this > information should be considered strictly confidential. If you have > received this email in error, please notify the sender immediately via a > return email for further direction. Thank you for your assistance. > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp From pam <@t> ategra.com Fri May 21 14:50:32 2004 From: pam <@t> ategra.com (Pam Barker (extension 234)) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Job Opportunities in Histology Latest Update 05/21/04 Message-ID: Hi Histonetters, Here is the latest update on opportunities with some of my best clients throughout the US who are seeking Histology Supervisors, Histo Technologists and Histo Technicians. These are positions as direct employees of our client. There are fulltime 40 hour per week positions.. As a direct employee of one of our clients you will be provided with full benefits including Health Insurance, Vacation, Sick Pay, Relocation money and a lucrative sign-on bonus. I have part time and temporary positions as well. I have supervisory, team lead and bench positions. These positions require HTL or HT certification or registry eligibility. Here are some of my HOTTEST Histology Supevisory positions: 1. Florida - Histology Supervisor 2. Alabama - Histology Supervisor 3. Missouri - Histology Supervisor 4. Oregon - Lead Tech (IHC) 5. New Hampshire - Histology Supervisor Here are some of my HOTTEST Histo Tech bench positions: 1. Florida - Histo Tech 2. Maine - Histo Tech 3. Georgia - Histo Tech 4. Pennsylvania - Histo Tech 5. Oregon - Histo Tech 6. Georgia - MOHS Tech 7. California - Immunohistochemistry Tech 8. Missouri - Histo Tech 9. Oregon - Histo Tech 10. Massachusetts - Histo Tech (part time) 11. Maryland - Histo Tech (part time) 12. Illinois - Histo Tech If you are interested in these jobs, please CALL ME ASAP at 800 466 9919 x234. To speed things up, please also send me a copy of your resume, (if you haven't already done so). If you are interested in jobs outside the above-mentioned areas, please send me your resume as well. I have clients throughout the US. I will keep your resume confidential and will not release it to anyone without your permission (This is Ategra policy as well as my own). My services are at no charge to you. Of course, you may be happy in your present job, but it never hurts to to keep an eye open. Also, if you have friends/peers who might be interested as well, if you could pass my query & name on to them I'd be very grateful. However, if you are interested in any of the jobs above, please call me. Thank You !! Pam - 800 466 9919 ext 234 --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing Learn More About Ategra: Pam Barker Senior Lab Recruiter Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd. Winter Park, FL 32792 VOICE: 407-671-5800 ext 234 TOLLFREE: 800-466-9919 ext 234 EMAIL: pam@ategra.com To Learn More About Ategra: 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 convmcm <@t> cc.usu.edu Fri May 21 15:35:01 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] I think Chloral Hydrate works better! In-Reply-To: <20040521.101613.24094.51760@webmail20.lax.untd.com> Message-ID: <000701c43f73$1735b080$4a737b81@Cygnus> How do you get Chloral hydrate? It's a controlled substance. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of madaryhtl@juno.com Sent: Friday, May 21, 2004 10:16 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] I think Chloral Hydrate works better! I make my own Mayers and have tried several recipes with so-called Chloral Hydrate substitutes and even no Chloral Hyrdate. I have done side-by-side studies(not published, there's an idea), and CHloral Hydrate does make a difference in the short run and in the long run if the stuff sits in the bottle longer than you expect. I say keep the Chloral Hydrate. Nick MAdary, B.S. HT/HTL(ASCP)QIHC Histology Manager Human Genome Sciences Rockville, Md 20850 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BMolinari <@t> heart.thi.tmc.edu Fri May 21 15:28:22 2004 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] unsubscribe Message-ID: From LINDA.MARGRAF <@t> childrens.com Fri May 21 15:53:45 2004 From: LINDA.MARGRAF <@t> childrens.com (LINDA MARGRAF) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Mesenteric node with obvious tumor Message-ID: Dear Sue Tell your pathologist about this great web site called immunoquery which will provide information about the reactivity of most any tumor with an encyclopedic number of antibodies. It comes complete with PubMed references and you can search by antibody or diagnosis. I find this website very helpful. It is run by Dr. Frisman who was (and maybe still is) a Histonet member. The address is www.ipox.org He will need to get a password to use the system. Good luck Linda M Histonet administrator Director of Anatomic Pathology Children's Medical Center of Dallas Associate Professor of Pathology Univ of Texas Southewestern Medical Center From myrtelina <@t> hotmail.com Fri May 21 16:45:45 2004 From: myrtelina <@t> hotmail.com (Myrtelina Fernandez) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] MarketLab Powdered Formalin Message-ID: I would like to know about other laboratories' experience with the use of this product. Our lab. its considering implementing its use. Thanks, M. Fernández Lab. Supervisor Myrtelina Fernández _________________________________________________________________ MSN 8 helps [1]ELIMINATE E-MAIL VIRUSES. Get 2 months FREE*. References 1. http://g.msn.com/8HMBEN/2752??PS=47575 From Carolyn.Earley <@t> leica-microsystems.com Fri May 21 17:12:50 2004 From: Carolyn.Earley <@t> leica-microsystems.com (Carolyn.Earley@leica-microsystems.com) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] (histonet)unsubscribe Message-ID: Carolyn Earley Marketing Manager Leica Microsystems 847-405-7014 Carolyn.Earley@Leica-Microsystems.com From madaryhtl <@t> juno.com Fri May 21 20:21:29 2004 From: madaryhtl <@t> juno.com (madaryhtl@juno.com) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] I think Chloral Hydrate works better! Message-ID: <20040521.182130.22945.57097@webmail14.lax.untd.com> I just order it from Fisher. I had to jump a few hoops with the company. They said I could just hang on to it in the lab, I just said keep it with the rest of the controlled stuff. I weigh out 200 g every time I make 4 liters. I think Jerry F even stated he thought it was better one time in a lecture on H&E's Do not quote me as he is a heavy player in that arena and I do not want to misquote. It would be like misquoting you! From TJJ <@t> Stowers-Institute.org Fri May 21 23:02:19 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] RE: Double skeletal stain on archived tissues Message-ID: I agree that it's possible your fixative has become acidic after one year's time and could be decalcifying your specimen. We did a simultaneous study using one adult mouse that had been fixed for over a year in 10% neutral buffered formalin, and one that had been freshly euthanized and processed according to our recommended protocol. We achieved staining on both, but the freshly processed sample was much more intense with much better quality than the previously fixed sample. Actually the archived tissue had very poor staining, nothing that was publishable or usable after we finished the technique. We have found that if the animal was placed and stored in a freezer soon after deat, there seems to be little to no compromise of the staining of these samples. We have not tested for long-term freezer storage however. We have not run into the problem of stain penetration on any of our mouse samples. We do remove the fat pad on the back of the neck on mice as part of the dissection and evisceration, and will also use an 8 hour - 1 day soak in acetone before starting the staining procedure. If you have access to an x-ray machine or faxitron, perhaps you can see if specimen decalcification is the problem here. Teri Johnson Stowers Institute for Medical Research Kansas City, MO From ekaplan <@t> squ.edu.om Sat May 22 05:39:16 2004 From: ekaplan <@t> squ.edu.om (Evelyn Kaplan) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] embalming course Message-ID: Good afternoon, Does anyone know of an emblaming course or a placement for 1-3 months please? I would apreciate some help if anyone can. Best wishes, Evelyn Kaplan, Dept of Pathology, College of Medicine and Health Sciences, Sultan Qaboos University, Oman From ekaplan <@t> squ.edu.om Sat May 22 06:09:26 2004 From: ekaplan <@t> squ.edu.om (Evelyn Kaplan) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] re Nordicware pressure cooker In-Reply-To: <000d01c435f8$e008ef60$40019b51@home> Message-ID: Good afternoon, I tried to buy this particular pressure cooker and was unable to place an overseas order. I also did not receive a reply to two emails I sent trying to see what the problem was. Can anyone help? Regards, Evelyn Kaplan, Dept of Pathology, College of Medicine and Health Sciences, Sultan Qaboos University, Oman -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Carl Sent: Saturday, May 08, 2004 11:26 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] re Nordicware pressure cooker Joe, I too use the Nordicware M/W pcooker; consistent , excellent results for the past 5yrs . Just interested in the solution/s you use to disclose your Ags.....Be grateful for info --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.668 / Virus Database: 430 - Release Date: 24/04/2004 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From peptolab <@t> hamptons.com Sat May 22 06:36:39 2004 From: peptolab <@t> hamptons.com (peptolab) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Hematoxylin Message-ID: <001101c43ff1$0d08b840$687bbd18@JEFF> When I was a teenage volunteer lab rat at the county medical center, I used to get all the scut work. I prepared big ehrlenmeyer flasks of Harris' hematoxylin that used to boil over when i added the mercuric oxide. I love the Gill formulations- especially Gill 3 in the frozen section set up- 30 seconds and you have perfect nuclei. In my career, Harris was always quirky, too much-not enough acid, strength erratically ofen rapidly deteriorated, gray nuclei, all kinds of muck forming daily like scum, sediment. With Gills- I need not filter at all most days, when a film forms, after about 5 or six days in my lab, its time to replace. Pretty much no matter the manufacturer, Gill hematoxylins are a pleasure to use and look at. Thanks Gary, for the invention. Jeff Silverman HT HTL QIHC (ASCP) Pathologists' Assistant Southside Hospital Bay Shore New York From hodges420 <@t> msn.com Sat May 22 20:45:31 2004 From: hodges420 <@t> msn.com (MARY T HODGES) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Alternative fixative users (Prefer) References: Message-ID: Thanks Pattie, I have seen first hand what happens with alcohol and ISH and Fish. IHC isn't the problem but when ISH or Fish was orded to back up ISH results. The outcome was a diaster. Thanks for bringing this out. Tere Hodges St. Mary's Hospital Tucson, Az ----- Original Message ----- From: Patti Loykasek To: histonet Sent: Wednesday, May 19, 2004 11:35 AM Subject: Re: [Histonet] Alternative fixative users (Prefer) I would caution any using Prefer to consider if they will be doing any rna/dna testing now or perhaps in the future on the tissue. We have had poor dna FISH results with it. Soon I'll be trying some pcr assays, and will let you know how that goes. Patti Loykasek PhenoPath Laboratories Seattle, WA > Hi, after many years as a Prefer fixative user our pathologists are telling > us that we have to go back to formalin. This came about after a letter to the > editor appeared in the Journal of Surgical Pathology. > > Has anyone out there been approached by their pathologists to switch back? > > Mary Bryhan > > > _______________________________________________ > 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 brokeponi <@t> yahoo.com Sun May 23 06:43:24 2004 From: brokeponi <@t> yahoo.com (Christine Baker) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Cutting Control Slides Message-ID: <20040523114324.53981.qmail@web60206.mail.yahoo.com> I am considering starting a control cutting service ot home. Does anyone know of anyone who does this?. I am going to need to know about the regs. etc about procuring blocks etc. I would like to market them to research and industry. This is in the very early research stages. I thought several years ago I heard of a company who would set you up and/or buy the slides from you. Thank you for any help you can offer. --------------------------------- Do you Yahoo!? Yahoo! Domains - Claim yours for only $14.70/year From Rcartun <@t> harthosp.org Sun May 23 08:30:27 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Cutting Control Slides Message-ID: You better check with your town/city public health department - they may have issues with you working on human/animal tissue at home (if they don't, your neighbors will). What are you going to do about waste disposal? Where are you planning on getting your tissue? Richard Cartun >>> Christine Baker 05/23/04 07:43AM >>> I am considering starting a control cutting service ot home. Does anyone know of anyone who does this?. I am going to need to know about the regs. etc about procuring blocks etc. I would like to market them to research and industry. This is in the very early research stages. I thought several years ago I heard of a company who would set you up and/or buy the slides from you. Thank you for any help you can offer. --------------------------------- Do you Yahoo!? Yahoo! Domains - Claim yours for only $14.70/year _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From qiaolin_deng78 <@t> hotmail.com Sun May 23 10:17:41 2004 From: qiaolin_deng78 <@t> hotmail.com (dengqiaolin deng) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] chicken in situ hybridization Message-ID: Dear histonetter: Is there anyone having done in situ hybridization for chicken cryosection? One kind of problems I have met is that the structure is easy to be crashed and disattached from the slide although I am very careful with every washing steps. But for the mouse cryosection, everything is fine. Is there any improvement that I can do to avoid it? Thank you for any suugestions in advance. Qiaolin Deng _________________________________________________________________ MSN 8 with [1]e-mail virus protection service: 2 months FREE* References 1. http://g.msn.com/8HMBEN/2740??PS=47575 From brucea <@t> unimelb.edu.au Mon May 24 01:23:00 2004 From: brucea <@t> unimelb.edu.au (Bruce Abaloz) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] GILL's Hematoxylin No.2....PLEASE Message-ID: I have searched 'high & low' (so to speak) & CANNOT find a recipe for Gill's Hematoxylin No. 2 . PLEASE - would some kind soul email me the recipe (to be used in the 'Charukian Method Oil Red O Dextrin Method)..........also, am after Nile Red to be used in the Spectrofluorometric studies of the lipid probe - Nile red.....is there 'another' name for Nile Red?? Cheers & THANKYOU very much in advance. Cheers, Bruce in OZ. -- BRUCE ABALOZ PH:61383446282 HISTOLOGIST FAX:61383447909 DEPT.of ZOOLOGY EMAIL: brucea@unimelb.edu.au THE UNIVERSITY Of MELBOURNE. VICTORIA.AUSTRALIA 3010 Nobody can make you feel inferior without your permission. - Eleanor Roosevelt DANCE LIKE NO-ONE'S WATCHING ******************************************************************************** This electronic message and all contents contain information which may be privileged, confidential or otherwise protected from disclosure.The information is intended to be for the addressee only. If you are not the addressee, any disclosure, copy, distribution or use of the contents of this message is prohibited. If you have received this electronic message in error, please notify us immediately and destroy the original message and all copies. From c.m.vanderloos <@t> amc.uva.nl Mon May 24 02:18:45 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] RE: Dead-End system & BMA/MMA embedded tissue Message-ID: <12048611cd49.11cd49120486@amc.uva.nl> Dear Sven, I can just confirm your observation: Once I tried to do a TUNEL staining on methacarn-fixed, paraffin-embedded(=similar to Carnoy, except the alcohol is here replaced by mehtanol) tissue along with standard formalin-fixed tonsil as positive control. The apoptotic nuclei in the formalin-fixed tissue were nicely stained but the staining of the methacarn-fixed tissue was terrible as you described. Apparently, the TUNEL staining is not fitting with this alcohol-based fixative. Hope this helps, Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Sven van Eijl Date Fri, 21 May 2004 17:28:39 +0100 To histonet@lists.utsouthwestern.edu Subject [Histonet] Dead-End system & BMA/MMA embedded tissue Dear Histonetters, This will be my first posting to this list, and I am by no means a real histologists, so bear with me please... We are having a problem using Promega's Dead-End colorimetric TUNEL kit on plastic embedded lungs. In the manual are only guidelines for formalin fixed and parrafin embedded or frozen tissue. We fixed the lungs using Carnoy's and embedded in BMA/MMA. I have had some tips from Promega about modifications to the protocol to account for the use of other fixatives than formalin, but am wondering if it is the fixative or the plastic that is causing the problems. We get a nice brown staining of all nuclei at all times, also in the negative controls. It is not a question of general high background, the rest of the tissue is clear. Any suggestions are highly appreciated. Thanks, Sven From mab70 <@t> medschl.cam.ac.uk Mon May 24 07:19:01 2004 From: mab70 <@t> medschl.cam.ac.uk (Margaret Blount) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Formalin fixation of cells for paraffin embedding? Message-ID: <6C21D947F7189448AB447C57BF2992B4025F16CE@mius.medlan.cam.ac.uk> Another cheap alternative is cigarette papers. Just leave out the contents! Margaret Margaret Blount Chief Technician Clinical Biochemistry University of Cambridge Addenbrooke's Hospital Hills Road Cambridge CB2 2QR -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bill Blank Sent: Friday, May 21, 2004 3:55 PM To: histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Formalin fixation of cells for paraffin embedding? At 8:52 AM -0500 5/21/04, Marsha Linville wrote: >Wrap the pellet in lens paper,and put it in a cassette to fix and process >with your tissues. We use those papers the hairdressers use when curling hair. Much cheaper than lens paper. Bill -- ______________ Bill Blank, MD Heartland Lab, Inc _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From L.D.Ridley <@t> bham.ac.uk Mon May 24 07:33:14 2004 From: L.D.Ridley <@t> bham.ac.uk (LD Ridley) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Brevican Message-ID: <1085401994.93865d20L.D.Ridley@bham.ac.uk> Hi All, Does anyone out there have any experience of using a Brevican antibody. Brevican is a member of the lectican family of proteoglycans expressed in the brain. We are trying to utilise this antibody in formalin fixed paraffin embedded brain tumour samples and would like to ask for some advice from anyone who has worked with Brevican. Thank you Lee Lee Ridley Research Associate L.D.Ridley@bham.ac.uk Cancer Research UK Institute for Cancer Studies University of Birmingham Vincent Drive Edgbaston Birmingham B15 2TT UK +44 (0)121 414 4472 From L.D.Ridley <@t> bham.ac.uk Mon May 24 07:36:58 2004 From: L.D.Ridley <@t> bham.ac.uk (LD Ridley) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Brevican Message-ID: <1085402218.93865d20L.D.Ridley@bham.ac.uk> Hi All, Does anyone out there have any experience of using a Brevican antibody. Brevican is a member of the lectican family of proteoglycans expressed in the brain. We are trying to utilise this antibody in formalin fixed paraffin embedded brain tumour samples and would like to ask for some advice from anyone who has worked with Brevican. Thank you Lee Lee Ridley Research Associate L.D.Ridley@bham.ac.uk Cancer Research UK Institute for Cancer Studies University of Birmingham Vincent Drive Edgbaston Birmingham B15 2TT UK +44 (0)121 414 4472 From rfail <@t> toolkitmail.com Mon May 24 07:52:02 2004 From: rfail <@t> toolkitmail.com (rfail) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] ALK-Phos staining of OCT? Message-ID: <40b1eff2.109.4c9b.1652605500@toolkitmail.com> Frozen sections stained with Mel-5 using an alkaline phosphatase are showing a pink blush surrounding the tissue on the slide. I washed the slides in running water for 30 minutes, then rinsed in buffer (TRIS), applied all our usual blocks, protein block, biotin blocking reagents, and levamisole, it's still there. The intensity is the same in the lower dilutions as in the higher. It does not interfere in the interpretation of the stain, but our docs are used to very "clean" IHC slides with both DAB and ALK-Phos procedures. Any help on how to get rid of this blush would be appreciated. Rena Fail, HT(ASCP) QIHC Medical University of South Carolina Charleston, SC From TJJ <@t> Stowers-Institute.org Mon May 24 08:40:16 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Re: chicken in situ hybridization Message-ID: Qiaolin, Are you using proteinase K enzyme digestion in your protocol? You may not need it on very young chick embryo cryosections. We've found that we don't need it most times on E2 whole mount ISH. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org Date: Sun, 23 May 2004 15:17:41 +0000 From: "dengqiaolin deng" Subject: [Histonet] chicken in situ hybridization To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain Dear histonetter: Is there anyone having done in situ hybridization for chicken cryosection? One kind of problems I have met is that the structure is easy to be crashed and disattached from the slide although I am very careful with every washing steps. But for the mouse cryosection, everything is fine. Is there any improvement that I can do to avoid it? Thank you for any suugestions in advance. Qiaolin Deng From bryand <@t> netbistro.com Mon May 24 08:46:02 2004 From: bryand <@t> netbistro.com (Bryan Llewellyn) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] GILL's Hematoxylin No.2....PLEASE In-Reply-To: References: Message-ID: <40B1FC9A.3090402@netbistro.com> All three are on StainsFile at: http://members.pgonline.com/~bryand/StainsFile/stain/hemalum/gill.htm Bryan Llewellyn Bruce Abaloz wrote: > I have searched 'high & low' (so to speak) & CANNOT find a recipe for > Gill's Hematoxylin No. 2 . PLEASE - would some kind soul email me the > recipe (to be used in the 'Charukian Method Oil Red O Dextrin > Method)..........also, am after Nile Red to be used in the > Spectrofluorometric studies of the lipid probe - Nile red.....is there > 'another' name for Nile Red?? Cheers & THANKYOU very much in advance. > Cheers, > > Bruce in OZ. From michael_lafriniere <@t> memorial.org Mon May 24 09:10:43 2004 From: michael_lafriniere <@t> memorial.org (LaFriniere, Mike) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Steven Slapp Info Message-ID: During the Region III Meeting this past weekend in Birmingham, I was informed that Steven Slapp was in a terrible accident. If anybody has information regarding Steven please contact me. Thank You Michael LaFriniere NSH Region III Director 423-495-6117 From amosbrooks <@t> earthlink.net Mon May 24 09:16:08 2004 From: amosbrooks <@t> earthlink.net (amosbrooks@earthlink.net) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] BCL6 w/o EDTA Message-ID: <1963539.1085408168926.JavaMail.root@bert.psp.pas.earthlink.net> Hi, If this helps, we use DAKO Hi pH retrieval solution in a steamer for 20 min. We use Envision for a detection kit as the Hi pH retrieval solution yeilds a lot of biotin staining. We use Novocastra primary ab at 1:5. Our results are good and if you use a really good charged slide and dry the sections well before staining the morphology is fine. good luck, Amos Brooks Message: 5 Date: Fri, 21 May 2004 13:46:27 -0400 From: Browning Deb Subject: [Histonet] BCL6, EDTA buffer To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <3AADFB88753AD31189C100902786B91C0E2781EC@hch_nt_exchange.hhsc.ca> Content-Type: text/plain; charset="iso-8859-1" Good afternoon: As much as I dislike EDTA buffer due to the poor morphology (especially if not properly fixed) that it renders, I have not had much success with BCL6 staining with any other buffers. I would appreciate any information on how to improve the morphology of the tissue, the use of other buffers, or any success stories, since I just can't seem to pull the rabbit out of the hat here. Thank you. From aglow <@t> wistar.upenn.edu Mon May 24 09:35:27 2004 From: aglow <@t> wistar.upenn.edu (Elsa Aglow) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] job posting Message-ID: The Wistar Institute is an independent, non-profit, medical research institute located on the University of Pennsylvania campus. We enjoy a reputation as one of the nation's oldest independent institutions devoted to basic biomedical research and training. Long known for our pioneering work and ground-breaking accomplishments in finding cures for diseases once thought incurable, we have been honored for the discovery of vaccines, genetic and molecular therapies and detection methods for disorders as varied as brain tumors, breast cancer, AIDS, Alzheimer's, multiple sclerosis and heart disease. We have three research programs: Gene Expression and Regulation, Molecular and Cellular Oncogenesis, and Immunology. The Wistar Institute has an immediate opening in our Histotechnology Core Facility. A Bachelor's degree with experience in all aspects of histologic technicques, in particular animal research histologic procedures, is required. Must have the ability to troubleshoot and multi-task independently on various projects. Good interpersonal skills are necessary as this position requires frequent interaction with research scientist. We offer an excellent benefits package, including tuition assistance. Apply to The Wistar Institute, HR Dept., 3601 Spruce St., Phila., PA 19104 EOE/AA/M/F/D/V. recruit@wistar.upenn.edu. For more information visit our website at www.wistar.upenn.edu. From qiaolin_deng78 <@t> hotmail.com Mon May 24 10:22:12 2004 From: qiaolin_deng78 <@t> hotmail.com (dengqiaolin deng) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Re: chicken in situ hybridization Message-ID: Hi, Teri: Thank you for your points. In my protocol, I treat the slides 5min with proteinase K, but the my chicken embryo is E4. Maybe I should shorten the time. Also, I want to ask how you do the hybridization step. In our lab, we drop 100ul hybridization solution with probe on the slide and then put a coverslip on it. The day after, we remove the coverslip by dipping the slides in the ssc solution. I find it is also a reason for losing the structure. Do you have other way to do it? Best regards Qiaolin >From: "Johnson, Teri" >To: >Subject: [Histonet] Re: chicken in situ hybridization >Date: Mon, 24 May 2004 08:40:16 -0500 > >Qiaolin, > >Are you using proteinase K enzyme digestion in your protocol? You may >not need it on very young chick embryo cryosections. We've found that >we don't need it most times on E2 whole mount ISH. > >Teri Johnson >Managing Director Histology Facility >Stowers Institute for Medical Research >1000 E. 50th St. >Kansas City, Missouri 64110 >tjj@stowers-institute.org > > > > >Date: Sun, 23 May 2004 15:17:41 +0000 >From: "dengqiaolin deng" >Subject: [Histonet] chicken in situ hybridization >To: histonet@lists.utsouthwestern.edu >Message-ID: >Content-Type: text/plain > > > Dear histonetter: > > Is there anyone having done in situ hybridization for >chicken > cryosection? One kind of problems I have met is that the structure >is > easy to be crashed and disattached from the slide although I am >very > careful with every washing steps. But for the mouse >cryosection, > everything is fine. Is there any improvement that I can do to >avoid > it? > Thank you for any suugestions in advance. > > Qiaolin Deng >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ The new [1]MSN 8: smart spam protection and 2 months FREE* References 1. http://g.msn.com/8HMBEN/2737??PS=47575 From histosci <@t> shentel.net Mon May 24 10:22:38 2004 From: histosci <@t> shentel.net (HSRL) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] CD31 Staining contract lab Message-ID: <000901c441a2$f79b6440$0200a8c0@HSRLMAIN> Dear Netters, We have a client who is need of CD31 staining. We do not offer this stain so I am looking to sub this out to a contract laboratory. Please e-mail me with your contact information. Thank you, Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 tomgalati@hsrl.org www.hsrl.org From TJJ <@t> Stowers-Institute.org Mon May 24 10:38:26 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Re: chicken in situ hybridization Message-ID: <1126709185-20541295@pathology.swmed.edu> Try lowering the concentration of proteinase K rather than the time. If you have an additional slide, try it with no pretreatment and see if that makes a difference. We remove the coverslip the same way, by dipping it into the SSC. If that isn't working for you, try just leaving the slides in SSC without the dipping motion and letting the coverslips just fall off on their own. How long do you let your cryosections air-dry before doing your protocol? And do you use any adhesive on your slide (+ charged, poly-l-lysine, etc.) Teri -----Original Message----- From: dengqiaolin deng [mailto:qiaolin_deng78@hotmail.com] Sent: Monday, May 24, 2004 10:22 AM To: Johnson, Teri Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: chicken in situ hybridization Hi, Teri: Thank you for your points. In my protocol, I treat the slides 5min with proteinase K, but the my chicken embryo is E4. Maybe I should shorten the time. Also, I want to ask how you do the hybridization step. In our lab, we drop 100ul hybridization solution with probe on the slide and then put a coverslip on it. The day after, we remove the coverslip by dipping the slides in the ssc solution. I find it is also a reason for losing the structure. Do you have other way to do it? Best regards Qiaolin From convmcm <@t> cc.usu.edu Mon May 24 10:36:36 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] GILL's Hematoxylin No.2....PLEASE In-Reply-To: <40B1FC9A.3090402@netbistro.com> Message-ID: <000c01c441a4$e681e150$4a737b81@Cygnus> Bryan, It's been some time since I visited the Stains File... thanks for the reminder! It's an awesome site. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn Sent: Monday, May 24, 2004 6:46 AM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] GILL's Hematoxylin No.2....PLEASE All three are on StainsFile at: http://members.pgonline.com/~bryand/StainsFile/stain/hemalum/gill.htm Bryan Llewellyn Bruce Abaloz wrote: > I have searched 'high & low' (so to speak) & CANNOT find a recipe for > Gill's Hematoxylin No. 2 . PLEASE - would some kind soul email me the > recipe (to be used in the 'Charukian Method Oil Red O Dextrin > Method)..........also, am after Nile Red to be used in the > Spectrofluorometric studies of the lipid probe - Nile red.....is there > 'another' name for Nile Red?? Cheers & THANKYOU very much in advance. > Cheers, > > Bruce in OZ. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Sarah.Blachford <@t> ngh.nhs.uk Mon May 24 10:48:12 2004 From: Sarah.Blachford <@t> ngh.nhs.uk (Blachford, Sarah - Histopath Main) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] dermatitis herpetiformis Message-ID: <854FA06D8BE2D611ABA30030052D1DF6FA58EA@NGH_EXCHANGE1> What antibodies do people use for diagnosing this. And do you use immunofluorescence or avidin biotin Sarah Northampton General Hospital NHS Trust Cliftonville, Northampton NN1 5BD This e-mail may contain confidential information and/or copyright material and is intended for the use of the addressee only. Any unauthorised use may be unlawful. The contents of this e-mail may be subject to public disclosure under the NHS Code of Openness or the Freedom of Information Act 2000. Unless legally exempt, the confidentiality of the message and your reply cannot be guaranteed. If you receive this e-mail by mistake, please advise the sender immediately. Thank you. From Terry.Marshall <@t> rothgen.nhs.uk Mon May 24 10:52:29 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] dermatitis herpetiformis Message-ID: The diagnostic immunologic pattern is granular IgA on the basement membrane region. Histologically the disease is identical to another in which the IgA is linear (called soundly enough, linear IgA disease), and very similar to bullous pemphigoid in which basement membrane IgG is found. Your panel should therefore include both. Immunofluorescence is unfortunately, the only way to go. 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: Blachford, Sarah - Histopath Main [mailto:Sarah.Blachford@ngh.nhs.uk] Sent: 24 May 2004 16:48 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] dermatitis herpetiformis What antibodies do people use for diagnosing this. And do you use immunofluorescence or avidin biotin Sarah Northampton General Hospital NHS Trust Cliftonville, Northampton NN1 5BD This e-mail may contain confidential information and/or copyright material and is intended for the use of the addressee only. Any unauthorised use may be unlawful. The contents of this e-mail may be subject to public disclosure under the NHS Code of Openness or the Freedom of Information Act 2000. Unless legally exempt, the confidentiality of the message and your reply cannot be guaranteed. If you receive this e-mail by mistake, please advise the sender immediately. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kspencer <@t> utmem.edu Mon May 24 11:11:11 2004 From: kspencer <@t> utmem.edu (Kathleen Spencer) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Steven Slapp Info In-Reply-To: Message-ID: I just heard that he is critical, multiple head injuries from a motorcycle accident. A good friend of his and co-worker is here at my lab today, her name is Dorothy Murphy, she is with Hacker Bright. She is upset because that is all she was told. kathleen On Monday, May 24, 2004, at 09:10 AM, LaFriniere, Mike wrote: > During the Region III Meeting this past weekend in Birmingham, I was > informed that Steven Slapp was in a terrible accident. If anybody has > information regarding Steven please contact me. > > Thank You > > Michael LaFriniere > NSH Region III Director > 423-495-6117 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Mon May 24 11:39:26 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Dead white Europeans Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3B38@exchsrv01.barrynet.barry.edu> Some classical music was composed by dead Americans, black (Scott Joplin) and white (Victor Herbert George Gershwin). 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 Gary Gill Sent: Tuesday, May 18, 2004 1:04 PM To: 'Connie McManus'; 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems You may have heard that classical music was composed by dead white Europeans. Well, Harris hematoxylin was composed by a dead white American (physician at Jefferson Hospital in 1904). Gill hematoxylin was composed by a live white American. So if you want to liven things up, go with Gill's! Gary Gill (one and the same) PS -- No royalties involved, thanks to bad advice in 1972 from corporate counsel for Johns Hopkins Medical School. -----Original Message----- From: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: Tuesday, May 18, 2004 10:41 AM To: 'Marshall Terry Dr,Consultant Histopathologist'; 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Wow. What a lot of interesting comments!! I agree with Terry re the agitation. When I watch the stainer do those dips (I can program how many, but NOT the briskness), I wonder if you could even call it agitation. My hand dips are very brisk. Also, I don't bother letting the slides stay in the alcohols for 1 minute or 2, I give the slides about 20 -30 good brisk dips in each solution, then the timed rinses & staining. This has always been far more satisfactory to me than those sllllooooowwwww dips from the machine. As for the kind of hematoxylin, someone suggested I throw out the Harris and do Gills III. I've tried Gill III before and I much prefer the Harris. So it's just a matter of personal preference on that... AND what your pathologist likes *G* Everyone having an nice Tuesday??? *g* Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, May 18, 2004 7:35 AM To: Connie McManus; Petia P Stefanova; Megan Kear; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems Connie remarks: "In truth, I prefer my hand stained sections better than when they're stained automatically." When I first saw what I call x-y stainers, I thought that we had in this, something that reproduced hand staining. Not so. What I think is lacking is the brisk agitation necessary to break down the interface between old and new solution. I'm not so sure about the rinsing either. 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: Connie McManus [mailto:convmcm@cc.usu.edu] Sent: 18 May 2004 15:09 To: 'Petia P Stefanova'; 'Megan Kear'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E stain problems We use almost the exact same protocol... we use Surgipath Harris, but we prepare eosin in-house. One thing I am amazed at in this protocol is the length of time in the acid alcohol. Do you use an autostainer? We have a Leica. The time is set to 1 second in the acid ETOH and sometimes the sections are almost too differentiated. I can't imagine 6 seconds in the acid ETOH!! Even when I do H&E manually, I dip the slides in and quickly put them in running water. I must have a very strong solution, I guess. Hmmmm. Interesting. In truth, I prefer my hand stained sections better than when they're stained automatically. Just wondering and blabbering (hey, it's Tuesday, what do you expect??) Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Petia P Stefanova Sent: Monday, May 17, 2004 6:45 AM To: Megan Kear; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] H&E stain problems Hi, I use Harris's hematoxylin which is also regressive and purchase my hematoxylin and eosin /alcohol-based/ from www.surgipath.com. I get very good H&E staining with this protocol. REAGENT TIME Xylene 3 min. Xylene 3 min. Abs. alc. 2 min. Abs. alc. 2 min 95% alc. 2 min 80% alc. 2 min Wash /tap water/ 30 sec. Hematoxylin 8 min. Wash /tap water/ 2 min. Acid alcohol 6 sec. Wash /tap water/ 2 min. Wash /tap water/ 5 min 80% alc. 30 sec. Eosin 15 sec. 95% alc. 10 sec. Abs. alc. 30 sec. Abs. alc. 30 sec. Xylene 1 min. Xylene 1 min. Xylene Exit Hope it helps! Petia _______________________________________________ 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 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 rfail <@t> toolkitmail.com Mon May 24 11:54:45 2004 From: rfail <@t> toolkitmail.com (rfail) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] RE: [IHCRG] ALK-PHOS> staining of OCT? Message-ID: <40b228d5.13e.27aa.766800564@toolkitmail.com> Mel-5 is for melanoma and stains melanocytes so DAB is not an option Have you tried using HRP instead of AP? I had a similar > problem with gut tissue on BVD (Bovine Viral Diarrhea) > except that in addition to this blush, I also got this > gorgeous red outline of all the microvilli even after > levamasole quenching... not what you want to see, but it's > artistically appealing. I concluded that gut is not a > good tissue to use with AP. There are some tissues that > have too much endogenous AP and quenching (blocking) just > isn't enough... you need to use a different detection > system. I don't really know if this is your problem or > not, but it's one thing you might want to consider. > > Connie McManus > Utah Veterinary Diagnostics Laboratory > Utah State University > Logan, UT > Phone: 435/797-1891 > fax: 435/797-2805 > > > -----Original Message----- > From: Rena Fail [mailto:rfail@charleston.net] > Sent: Monday, May 24, 2004 2:37 AM > To: histonet@lists.utsouthwetern.edu; Ihcrg > Subject: [IHCRG] ALK-PHOS> staining of OCT? > > Frozen sections stained with Mel-5 using an alkaline > phosphatase procedure are showing a pink blush surrounding > the tissue on the slide.I washed the slides in running > water for 30 minutes, then rinsed in buffer (TRIS), > applied all our usual blocks, protein block, biotin > blocking reagents, and levamisole, it's still there. The > intensity is exactly the same in the lower dilutions as in > the higher. It doesn't interfere with the interpretation > of the stain, but our docs are used to very 'clean" IHC > slides for DAB and ALK-Phos. I used treated slides. Any > advice on how to get rid of this blush would be > appreciated. > > Rena Fail, HT(ASCP)QIHC > IHC/SS > Medical University of SC > 165 Ashley Ave. > Charleston, SC 29425 > > > > [Non-text portions of this message have been removed] > > > > > Yahoo! Groups Links > > > > > > > > ------------------------ Yahoo! Groups Sponsor > ---------------------~--> Make a clean sweep of pop-up > ads. Yahoo! Companion Toolbar. Now with Pop-Up Blocker. > Get it for free! > http://us.click.yahoo.com/L5YrjA/eSIIAA/yQLSAA/asSolB/TM > ---------------------------------------------------------- > -----------~-> > > > Yahoo! Groups Links > > <*> To visit your group on the web, go to: > http://groups.yahoo.com/group/IHCRG/ > > <*> To unsubscribe from this group, send an email to: > IHCRG-unsubscribe@yahoogroups.com > > <*> Your use of Yahoo! Groups is subject to: > http://docs.yahoo.com/info/terms/ > > From convmcm <@t> cc.usu.edu Mon May 24 12:11:18 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] RE: [IHCRG] ALK-PHOS> staining of OCT? In-Reply-To: <40b228d5.13e.27aa.766800564@toolkitmail.com> Message-ID: <001201c441b2$20e15940$4a737b81@Cygnus> You don't need to use DAB... you can use AEC. It has a nice dark red stain. This is soluble in ETOH and xylene, so you need to use an aqueous mounting media. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: rfail@toolkitmail.com [mailto:rfail@toolkitmail.com] Sent: Monday, May 24, 2004 9:55 AM To: Connie McManus Cc: histonet@lists.utsouthwestern.edu Subject: RE: [IHCRG] ALK-PHOS> staining of OCT? Mel-5 is for melanoma and stains melanocytes so DAB is not an option Have you tried using HRP instead of AP? I had a similar > problem with gut tissue on BVD (Bovine Viral Diarrhea) > except that in addition to this blush, I also got this > gorgeous red outline of all the microvilli even after > levamasole quenching... not what you want to see, but it's > artistically appealing. I concluded that gut is not a > good tissue to use with AP. There are some tissues that > have too much endogenous AP and quenching (blocking) just > isn't enough... you need to use a different detection > system. I don't really know if this is your problem or > not, but it's one thing you might want to consider. > > Connie McManus > Utah Veterinary Diagnostics Laboratory > Utah State University > Logan, UT > Phone: 435/797-1891 > fax: 435/797-2805 > > > -----Original Message----- > From: Rena Fail [mailto:rfail@charleston.net] > Sent: Monday, May 24, 2004 2:37 AM > To: histonet@lists.utsouthwetern.edu; Ihcrg > Subject: [IHCRG] ALK-PHOS> staining of OCT? > > Frozen sections stained with Mel-5 using an alkaline > phosphatase procedure are showing a pink blush surrounding > the tissue on the slide.I washed the slides in running > water for 30 minutes, then rinsed in buffer (TRIS), > applied all our usual blocks, protein block, biotin > blocking reagents, and levamisole, it's still there. The > intensity is exactly the same in the lower dilutions as in > the higher. It doesn't interfere with the interpretation > of the stain, but our docs are used to very 'clean" IHC > slides for DAB and ALK-Phos. I used treated slides. Any > advice on how to get rid of this blush would be > appreciated. > > Rena Fail, HT(ASCP)QIHC > IHC/SS > Medical University of SC > 165 Ashley Ave. > Charleston, SC 29425 > > > > [Non-text portions of this message have been removed] > > > > > Yahoo! Groups Links > > > > > > > > ------------------------ Yahoo! Groups Sponsor > ---------------------~--> Make a clean sweep of pop-up > ads. Yahoo! Companion Toolbar. Now with Pop-Up Blocker. > Get it for free! > http://us.click.yahoo.com/L5YrjA/eSIIAA/yQLSAA/asSolB/TM > ---------------------------------------------------------- > -----------~-> > > > Yahoo! Groups Links > > <*> To visit your group on the web, go to: > http://groups.yahoo.com/group/IHCRG/ > > <*> To unsubscribe from this group, send an email to: > IHCRG-unsubscribe@yahoogroups.com > > <*> Your use of Yahoo! Groups is subject to: > http://docs.yahoo.com/info/terms/ > > From Rcartun <@t> harthosp.org Mon May 24 12:16:58 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] dermatitis herpetiformis Message-ID: Immunofluorescence (specifically IgA, C3, and fibrin) on frozen tissue. Richard Cartun >>> "Blachford, Sarah - Histopath Main" 05/24/04 11:48AM >>> What antibodies do people use for diagnosing this. And do you use immunofluorescence or avidin biotin Sarah Northampton General Hospital NHS Trust Cliftonville, Northampton NN1 5BD This e-mail may contain confidential information and/or copyright material and is intended for the use of the addressee only. Any unauthorised use may be unlawful. The contents of this e-mail may be subject to public disclosure under the NHS Code of Openness or the Freedom of Information Act 2000. Unless legally exempt, the confidentiality of the message and your reply cannot be guaranteed. If you receive this e-mail by mistake, please advise the sender immediately. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From convmcm <@t> cc.usu.edu Mon May 24 12:18:44 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Steven Slapp Info In-Reply-To: Message-ID: <001401c441b3$2b3a9ef0$4a737b81@Cygnus> This is awful!! Can we send him some kind of Histonet greeting card? Please try to keep us informed on how he's doing. Here's hoping he's back with us soon. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kathleen Spencer Sent: Monday, May 24, 2004 9:11 AM To: LaFriniere, Mike Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Steven Slapp Info I just heard that he is critical, multiple head injuries from a motorcycle accident. A good friend of his and co-worker is here at my lab today, her name is Dorothy Murphy, she is with Hacker Bright. She is upset because that is all she was told. kathleen On Monday, May 24, 2004, at 09:10 AM, LaFriniere, Mike wrote: > During the Region III Meeting this past weekend in Birmingham, I was > informed that Steven Slapp was in a terrible accident. If anybody has > information regarding Steven please contact me. > > Thank You > > Michael LaFriniere > NSH Region III Director > 423-495-6117 > _______________________________________________ > 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 web <@t> histosoft.com Mon May 24 13:11:19 2004 From: web <@t> histosoft.com (web@histosoft.com) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Get Well Steven! Message-ID: <1085422279.40b23ac7818ba@webmail2.catalog.com> If the reports of Steven Slap's accident are true, then we are deeply saddened by the incident. HistoSoft would like to wish Steven a rapid recovery and condolences to EM Science and related families for the unfortunate circumstances. I know this speaks for many when we say "GET WELL SOON, Steven!". Best wishes to a rapid recovery, Todd Todd Sherman President HistoSoft Corporation "Biology in a new form..." Home: www.histosoft.com Member Services: www.myhistosoft.com From mariatere <@t> infovia.com.ar Mon May 24 13:20:19 2004 From: mariatere <@t> infovia.com.ar (mariatere@infovia.com.ar) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Unsuscribe, please Message-ID: <293580-220045124182019146@M2W103.mail2web.com> Please unsuscribe me for a while. I will be on holidays from 29th May , to 13th July. Ht. Maria Teresa Dominguez Anatomic Pathology Service Hospital Regional R?o Grande. Tierra del Fuego, Argentina. -------------------------------------------------------------------- mail2web - Check your email from the web at http://mail2web.com/ . From kspencer <@t> utmem.edu Mon May 24 13:22:55 2004 From: kspencer <@t> utmem.edu (Kathleen Spencer) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Steven Slapp Info In-Reply-To: <001401c441b3$2b3a9ef0$4a737b81@Cygnus> Message-ID: <60A303EB-ADAF-11D8-9D9B-000393967904@utmem.edu> I spoke with Dorothy again. She said she tried to call him at the hospital and he could not recieve calls, and someone called his father in California and he told them that Steven was not expected to make it. Sorry to bring bad news. -Kathleen On Monday, May 24, 2004, at 12:18 PM, Connie McManus wrote: > This is awful!! Can we send him some kind of Histonet greeting card? > Please try to keep us informed on how he's doing. > Here's hoping he's back with us soon. > > Connie McManus > Utah Veterinary Diagnostics Laboratory > Utah State University > Logan, UT > Phone: 435/797-1891 > fax: 435/797-2805 > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kathleen > Spencer > Sent: Monday, May 24, 2004 9:11 AM > To: LaFriniere, Mike > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Steven Slapp Info > > I just heard that he is critical, multiple head injuries from a > motorcycle accident. A good friend of his and co-worker is here at my > lab today, her name is Dorothy Murphy, she is with Hacker Bright. She is > > upset because that is all she was told. > > kathleen > On Monday, May 24, 2004, at 09:10 AM, LaFriniere, Mike wrote: > >> During the Region III Meeting this past weekend in Birmingham, I was >> informed that Steven Slapp was in a terrible accident. If anybody has >> information regarding Steven please contact me. >> >> Thank You >> >> Michael LaFriniere >> NSH Region III Director >> 423-495-6117 >> _______________________________________________ >> 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 JWEEMS <@t> sjha.org Mon May 24 13:40:28 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Steven Slapp Info Message-ID: Can you get word to him that our spirits are with him. He can be told that anyway. This is terrible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Kathleen Spencer Sent: Monday, May 24, 2004 2:23 PM To: Connie McManus Cc: histonet@lists.utsouthwestern.edu; 'LaFriniere,Mike' Subject: Re: [Histonet] Steven Slapp Info I spoke with Dorothy again. She said she tried to call him at the hospital and he could not recieve calls, and someone called his father in California and he told them that Steven was not expected to make it. Sorry to bring bad news. -Kathleen On Monday, May 24, 2004, at 12:18 PM, Connie McManus wrote: > This is awful!! Can we send him some kind of Histonet greeting card? > Please try to keep us informed on how he's doing. > Here's hoping he's back with us soon. > > Connie McManus > Utah Veterinary Diagnostics Laboratory > Utah State University > Logan, UT > Phone: 435/797-1891 > fax: 435/797-2805 > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kathleen > Spencer > Sent: Monday, May 24, 2004 9:11 AM > To: LaFriniere, Mike > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Steven Slapp Info > > I just heard that he is critical, multiple head injuries from a > motorcycle accident. A good friend of his and co-worker is here at my > lab today, her name is Dorothy Murphy, she is with Hacker Bright. She is > > upset because that is all she was told. > > kathleen > On Monday, May 24, 2004, at 09:10 AM, LaFriniere, Mike wrote: > >> During the Region III Meeting this past weekend in Birmingham, I was >> informed that Steven Slapp was in a terrible accident. If anybody has >> information regarding Steven please contact me. >> >> Thank You >> >> Michael LaFriniere >> NSH Region III Director >> 423-495-6117 >> _______________________________________________ >> 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 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 EMarquez <@t> crch.hawaii.edu Mon May 24 14:02:54 2004 From: EMarquez <@t> crch.hawaii.edu (Eddie Marquez) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] unsuscribe Message-ID: please take me off the list for a while, I'll be off island for sometime. Thank You eddie M From myrtelina <@t> hotmail.com Mon May 24 14:48:17 2004 From: myrtelina <@t> hotmail.com (Myrtelina Fernandez) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] MarkeLab Powdered Formalin Message-ID: Our lab its considering implementing the use of powdered formalin, Do you have any experience with the use of this product? Thanks, M Fernández Lab. Supervisor Myrtelina Fernández _________________________________________________________________ Tired of spam? Get [1]advanced junk mail protection with MSN 8. References 1. http://g.msn.com/8HMBEN/2734??PS=47575 From Gillian.Barlow <@t> cshs.org Mon May 24 15:01:15 2004 From: Gillian.Barlow <@t> cshs.org (Barlow, Gillian) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] unsubscribe Message-ID: <3CFAA0108952D111A5BF00805FA6FB0F079475B4@PEDSNTAS.csmc.edu> please unsubscribe me as I will be on medical leave for some time Many thanks Gillian Gillian M. Barlow, PhD Postdoctoral Fellow Laboratory of Julie Korenberg, PhD, MD Cedars-Sinai Medical Center Davis Bldg, Lab 2007 110 George Burns Rd Los Angeles, CA 90048 Phone: (310) 423 7650 Fax: (310) 423 0302 From DMBCMP <@t> aol.com Mon May 24 16:01:25 2004 From: DMBCMP <@t> aol.com (DMBCMP@aol.com) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] TO STEVEN and FAMILY Message-ID: <42.4ec75fe7.2de3bca5@aol.com> Dear Steven: The Histology Department at Fresno Community Hospital, Fresno California, want you to know that our thoughts are with you. We are deeply saddened. Dannie Blake From asmith <@t> mail.barry.edu Mon May 24 17:48:45 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Steven Slapp Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3B39@exchsrv01.barrynet.barry.edu> My grandfather's doctor told him at 80 that he had 6 months to live: he lived another 13 years. I hope your doctor is as wrong as my grandfather's. With heartfelt wishes for your recovery, Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 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 mrsgbd2001 <@t> yahoo.com Mon May 24 18:09:00 2004 From: mrsgbd2001 <@t> yahoo.com (Gareth Davis) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Re: rat turbinates In-Reply-To: <3.0.6.32.20040524082517.00c44e68@gemini.msu.montana.edu> Message-ID: <20040524230900.59323.qmail@web13310.mail.yahoo.com> Hello all, Superfrost Plus slides are wonderful. So far anyway. I successfully H&E stained my rat turbinate cryosections (after acetone/alcohol fixation), have not tried immunohistochemistry yet. Thanks to everyone for your help. Gareth Davis Research Assistant Tennessee State University Nashville, Tennessee Gayle Callis wrote: 5 micrometers without problems, but I do cut sucrose cryoprotected at -25 to -26C, and use AccuEdge high profile, some knives are pure junk! At 03:49 PM 5/14/2004 -0700, you wrote: >Gayle, Just wanted to get your opinion. Thanks, Gareth Do you Yahoo!? >SBC Yahoo! - Internet access at a great low price. 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) --------------------------------- Do you Yahoo!? Friends. Fun. Try the all-new Yahoo! Messenger From JColCLEFA <@t> aol.com Mon May 24 18:14:08 2004 From: JColCLEFA <@t> aol.com (JColCLEFA@aol.com) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] BCL6 Message-ID: I use dako's Bcl6 @1:20, Biogenex Supersensitive concentrated HRP/DAB kit, Cellmarque's trilogy for 25 Min @ 98C and get good staining, no BG and minimal disruption from EDTA although It's an obvious difference from non EDTA retrieval. From malcolm_gillham <@t> health.qld.gov.au Mon May 24 19:10:51 2004 From: malcolm_gillham <@t> health.qld.gov.au (Malcolm Gillham) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] unsubscribe Message-ID: Please unsubscribe me *********************************************************************************** This email, including any attachments sent with it, is confidential and for the sole use of the intended recipient(s). This confidentiality is not waived or lost, if you receive it and you are not the intended recipient(s), or if it is transmitted/received in error. Any unauthorised use, alteration, disclosure, distribution or review of this email is prohibited. It may be subject to a statutory duty of confidentiality if it relates to health service matters. If you are not the intended recipient(s), or if you have received this email in error, you are asked to immediately notify the sender by telephone or by return email. You should also delete this email and destroy any hard copies produced. *********************************************************************************** From anissachoi <@t> hotmail.com Mon May 24 21:59:28 2004 From: anissachoi <@t> hotmail.com (Anissa Choi) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Immuno slide problem Message-ID: We run our immuno on Ventana Nexus IHC. We place a positive control section on every slide together with patient 's tissue section. We use charged slides with a red edge square which is right next to the barcode label . The square is labelled "control" and that is where we place the positive control section. We had a case of a punch skin bx on which the pathologist requested LCA, L26 and CD3. Because of the small size of specimen, 2 sections were picked up on each slide. There was no problem with LCA, the positive control and the 2 skin sections stained strongly . For L26, the control worked but only 1 skin section ,which was at the bottom end of the slide, was immunostained strongly for L26. The other skin section in the middle and lying next to the positive control was completely negative for L26 but nuclei were stained with hematoxylin. The same staining pattern was seen in CD3-----the skin section in the middle did not get stained while the control and the other section worked just fine. Ventana suggested us to switch to other kind of slides, saying some stuff from the red edge may get onto the slides and cause staining problem. We requested a maintenance check on the stainer and Venatana could not find anything wrong with the machine. We recut the section using same kind of slides and repeated L26 and CD3. We got normal result with both skin sections stained . We also re-run the origional problem slides on Ventana but they turned out the same, the skin section in the middle still not stained. I am convinced that there is something on the slides that is blocking the staining but I am puzzled by the fact that the nuclei are stained . I would like to get some comments or opinions on this. I also hope to hear from people using the same type of slides to see if they have any staining problems. Thanks in advance for your inputs. Anissa Choi _________________________________________________________________ STOP MORE SPAM with the MSN Premium and get 2 months FREE* http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines From soumya25 <@t> nbrc.ac.in Mon May 24 22:55:12 2004 From: soumya25 <@t> nbrc.ac.in (soumya) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Unsubscribe Message-ID: <006501c4420c$15303c60$3e00a8c0@nbrc.btisnet.ac.in> Please unsubscribe me from the Histonet mailing list. Thanks, Dr. Soumya Iyengar Scientist National Brain Research Centre, NH-8, Manesar, Haryana - 122050, India Ph. no. 0124-2338922-8926 From t-sherman <@t> comcast.net Tue May 25 01:05:37 2004 From: t-sherman <@t> comcast.net (Todd Sherman) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Immuno slide problem Message-ID: <40B2E231.2060802@comcast.net> -----BEGIN PGP SIGNED MESSAGE----- Hash: SHA1 Anissa, I have not used the Ventana Nexus IHC or any slides of the type you describe, but I have a question for your QC follow-up. What happens when you use only positive control sections on the slide, i.e. a row of positive sections only? Do you observe the same unequal distribution of positive staining in a similar (or for that matter, different) pattern? If I understand you correctly, you observed the following: (Hopefully this schematic will resolve properly in your email reader. It is in ASCII format so you should be able to cut, copy, and paste this graphic representation on your computers notepad editor.) ~ Label +Control Specimen L Specimen R +-------+------------+-----------------------------+ | | +Con LCA + | LCA + LCA + | | | | | | | +Con L26 + | L26 - L26 + | | | | | | | +Con CD3 + | CD3 - CD3 + | +-------+------------+-----------------------------+ ~ == ==== ~ ==== ======== ~ ====== ============ ~ ======== ================ ~ ========== ==================== ~ Red "Box" Specimen Area ~ Hematoxylin+ Hematoxylin+ Notes: + indicates positive immunodetection - - indicates negative immunodetection = indicates levels of polarized IHC staining via histogram (more bars equates to higher intensity) It would seem to me, as you have alluded, that the red barrier material is acting as a sort of hydrophobic barrier (like a PAP pen) or a physical one whereby a slightly tilted slide tray might cause pooling of liquid volumes. The red "raised" barrier could impede the natural flow of liquid to the far end (label terminal vs label proximal) of the slide and allow the polar staining pattern that you see. If you mount a row of positive control sections on the same slide, immunostain as before, and observe the same pattern, it implicates the slide. If you change slides (i.e. sans red barrier), mount the same row of positive control sections, and observe only positive staining near the terminal end (opposite label), then that implicates the immunostainer or possibly the tray. It might be out of balance or tilted. Since I'm not familiar with your staining procedure, this is just troubleshooting. Incubation times for immunoreagents are typically much longer than for hematoxylin, so the sections will stain much more readily for hematoxylin. It wouldn't surprise me that hematoxylin staining works while IHC detection doesn't work because of the nature of the chemical reactions and thermodynamics that are occurring. An adequate volume of reagent must coat the entire section at all times, as you know, so a bi-polar pooling of reagents exacerbated by that "red barrier" could explain your staining patterns. Let us know what happens with all positive controls on a slide and let the experts consult. Hope this helps, Todd Todd Sherman President HistoSoft Corporation "Biology in a new form..." Home: www.histosoft.com Member Services: www.myhistosoft.com ________________________________________________________________________ From: "Anissa Choi" We run our immuno on Ventana Nexus IHC. We place a positive control section on every slide together with patient 's tissue section. We use charged slides with a red edge square which is right next to the barcode label. The square is labelled "control" and that is where we place the positive control section. We had a case of a punch skin bx on which the pathologist requested LCA, L26 and CD3. Because of the small size of specimen, 2 sections were picked up on each slide. There was no problem with LCA, the positive control and the 2 skin sections stained strongly. For L26, the control worked but only 1 skin section ,which was at the bottom end of the slide, was immunostained strongly for L26. The other skin section in the middle and lying next to the positive control was completely negative for L26 but nuclei were stained with hematoxylin. The same staining pattern was seen in CD3-----the skin section in the middle did not get stained while the control and the other section worked just fine. Ventana suggested us to switch to other kind of slides, saying some stuff from the red edge may get onto the slides and cause staining problem. We requested a maintenance check on the stainer and Venatana could not find anything wrong with the machine. We recut the section using same kind of slides and repeated L26 and CD3. We got normal result with both skin sections stained. We also re-run the origional problem slides on Ventana but they turned out the same, the skin section in the middle still not stained. I am convinced that there is something on the slides that is blocking the staining but I am puzzled by the fact that the nuclei are stained. I would like to get some comments or opinions on this. I also hope to hear from people using the same type of slides to see if they have any staining problems. Thanks in advance for your inputs. Anissa Choi -----BEGIN PGP SIGNATURE----- Version: GnuPG v1.2.3 (MingW32) Comment: Using GnuPG with MultiZilla - http://enigmail.mozdev.org iD8DBQFAsuIwEmHXdrslGRcRAhX5AKDBFrtQ3/3FJ2kVNdi9HFXtZvqnYgCghUP4 0QaUjbw2EpOePUYnbY3lqVE= =pwvT -----END PGP SIGNATURE----- From qiaolin_deng78 <@t> hotmail.com Tue May 25 02:32:07 2004 From: qiaolin_deng78 <@t> hotmail.com (dengqiaolin deng) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] RE:Re: chicken in situ hybridization Message-ID: Hi, Teri: Thank you very much for your further suggestions. As to the slides, we use SuperFrost Plus slides which is ready to use. So I don't do anything to the slides. Before starting in situ, I only air-dry the slides briefly. Actully, I didn't realizae it is an important step before you point it out. How long do you usually do? Qiaolin >From: "Johnson, Teri" >To: "dengqiaolin deng" >CC: >Subject: RE: [Histonet] Re: chicken in situ hybridization >Date: Mon, 24 May 2004 10:38:26 -0500 > >Try lowering the concentration of proteinase K rather than the time. If >you have an additional slide, try it with no pretreatment and see if >that makes a difference. We remove the coverslip the same way, by >dipping it into the SSC. If that isn't working for you, try just >leaving the slides in SSC without the dipping motion and letting the >coverslips just fall off on their own. How long do you let your >cryosections air-dry before doing your protocol? And do you use any >adhesive on your slide (+ charged, poly-l-lysine, etc.) > >Teri > > -----Original Message----- > From: dengqiaolin deng [mailto:qiaolin_deng78@hotmail.com] > Sent: Monday, May 24, 2004 10:22 AM > To: Johnson, Teri > Cc: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Re: chicken in situ hybridization > > > > Hi, Teri: > > > Thank you for your points. In my protocol, I treat the slides >5min with proteinase K, but the my chicken embryo is E4. Maybe I should >shorten the time. Also, I want to ask how you do the hybridization step. >In our lab, we drop 100ul hybridization solution with probe on the slide >and then put a coverslip on it. The day after, we remove the coverslip >by dipping the slides in the ssc solution. I find it is also a reason >for losing the structure. Do you have other way to do it? > > Best regards > > Qiaolin > > > > _________________________________________________________________ Add photos to your e-mail with [1]MSN 8. Get 2 months FREE*. References 1. http://g.msn.com/8HMAEN/2746??PS=47575 From h.newbery <@t> ed.ac.uk Tue May 25 04:14:26 2004 From: h.newbery <@t> ed.ac.uk (Helen Newbery) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Spleen autofluorescence Message-ID: <1085476466.40b30e723e67f@staffmail.ed.ac.uk> Hi histonetters, Apologies if my query is really basic but I am wanting to do immunofluorescent antibody staining on mouse spleen sections (paraffin-embedded) and am seeing a lot of autofluorescence, which I presume is due to platelets. Is there any way around this problem? Helen Newbery, Medical Genetics Section, Molecular Medicine Centre, Western General Hospital, Edinburgh, EH4 2XU. 0131 651 1047 From mab70 <@t> medschl.cam.ac.uk Tue May 25 05:28:20 2004 From: mab70 <@t> medschl.cam.ac.uk (Margaret Blount) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Steven Slapp Info Message-ID: <6C21D947F7189448AB447C57BF2992B4025F16D3@mius.medlan.cam.ac.uk> I add my good wishes for Steven's rapid recovery. Margaret Margaret Blount Chief Technician Clinical Biochemistry University of Cambridge Addenbrooke's Hospital Hills Road Cambridge CB2 2QR -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Connie McManus Sent: Monday, May 24, 2004 6:19 PM To: 'Kathleen Spencer'; 'LaFriniere, Mike' Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Steven Slapp Info This is awful!! Can we send him some kind of Histonet greeting card? Please try to keep us informed on how he's doing. Here's hoping he's back with us soon. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kathleen Spencer Sent: Monday, May 24, 2004 9:11 AM To: LaFriniere, Mike Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Steven Slapp Info I just heard that he is critical, multiple head injuries from a motorcycle accident. A good friend of his and co-worker is here at my lab today, her name is Dorothy Murphy, she is with Hacker Bright. She is upset because that is all she was told. kathleen On Monday, May 24, 2004, at 09:10 AM, LaFriniere, Mike wrote: > During the Region III Meeting this past weekend in Birmingham, I was > informed that Steven Slapp was in a terrible accident. If anybody has > information regarding Steven please contact me. > > Thank You > > Michael LaFriniere > NSH Region III Director > 423-495-6117 > _______________________________________________ > 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 Hitesh.Asarpota <@t> med.ge.com Tue May 25 07:00:27 2004 From: Hitesh.Asarpota <@t> med.ge.com (Asarpota, Hitesh (MED, Intern)) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Cost of Ventana Benchmark XT? Message-ID: <6EDD9DC298BF9C48AC3EF66EA92596BC14E398E4@frbucmsx03medge> Hello, Does someone know the approx cost of Cost of Ventana Benchmark XT in the U.S ? Tx, Hitesh Asarpota ---------------------------------------------------------- GE HealthCare Business Development Tel: +33 01 30 70 9224 Cell: +33 06 1171 4370 ---------------------------------------------------------- From asmith <@t> mail.barry.edu Tue May 25 08:50:59 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Retic nomenclature Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3B3A@exchsrv01.barrynet.barry.edu> These days, "reticulin" is usually used to designate a specific tissue component, viz., type III collagen. Type III collagen usually stains black with reticulin stains. Type I collagen usually stains brown. Laidlaw's diammine silver carbonate gives the best results in my hands. Although the old tinctorial and metal precipitation methods are not quite as specific as immunohistochemical methods, they cost about 1/10 th as much for materials and usually take less time to perform. 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 RSRICHMOND@aol.com Sent: Saturday, May 15, 2004 2:24 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Retic nomenclature Lance Erickson at Primary Children's Medical Center in Salt Lake City Utah asks about the appropriate name for the reticulum stain, Gordon and Sweets or any other. The historically sanctioned name is reticulum stain, and there is no reason to change it. Thirty or forty years ago "reticulin" was sometimes considered to be a specific tissue component that the stain demonstrated, but since this is no longer thought to be the case the word "reticulin" should be abandoned. - Supposedly the PAS stain has similar specificity. Reticulum stains go back to the Chandler Foot era of surgical pathology (1930's-early 1940's), when it was thought that special stains would revolutionize surgical pathology in the way that immunohistochemistry actually did in the 1980's. By the late 1960's the stain was out of use in many large centers; I don't think I ever saw one when I was at Johns Hopkins between 1964 and 1972. I always thought of it as an old codger's stain when I was young. Now that I AM an old codger, I'm surprised it's still around. Bob Richmond Samurai Pathologist Gastonia NC _______________________________________________ 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 MinHan.Tan <@t> vai.org Tue May 25 10:09:08 2004 From: MinHan.Tan <@t> vai.org (Tan, MinHan) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Spleen autofluorescence Message-ID: <74D0F0AB07F2E647A02D839ED79520F9C233DD@VAIEXCH02.vai.org> If you think it is autofluorescence due to tissue, which is possible, view it before adding any antibody. Actually, I think formalin-fixed tissue tends to have high autofluorescence. I attach here a post which I found on Histonet. From: "J. A. Kiernan" ------------------------------------------------------------------------ -------- Entering autofluorescence as a search term at http://www.histosearch.com brought up 100 hits all dated in the last 3 years. This is a problem many people have, and the number and variety of remedies indicate that there's no perfect solution. In one recent comparative study, Baschong, Suetterlin and Laeng (J. Histochem. Cytochem. 49:1565-1571, 2001) found that staining for 30 min with 0.1% sudan black B in 70% alcohol, after immunofluorescent staining, solved most autofluorescence problems. After staining the dye was washed off with a jet of a modified Hanks's solution and then immersed in the same solution for 10 minutes. The authors said this washing was necessary to avoid formation of a black precipitate. John Kiernan London, Canada ----------------------------------- "Hines, Edith M." wrote: > > > I am having some difficulty overcoming background staining on > > formalin-fixed, paraffin-embedded, human lung tissue sections. I have > tried blocking > > the tissues with chromotrope 2R, and with ammonium chloride. Both of > these > > methods have not significantly reduced the amount of autofluorescence that > I > > observe. > > Any advice that may be able to provide would be greatly > > appreciated. > > > > Thank you for your time, > > Edie Hines > > Research Technologist > > Mayo Clinic Scottsdale > > hines.edith@mayo.edu > > -----Original Message----- From: Helen Newbery [mailto:h.newbery@ed.ac.uk] Sent: Tuesday, May 25, 2004 5:14 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Spleen autofluorescence Hi histonetters, Apologies if my query is really basic but I am wanting to do immunofluorescent antibody staining on mouse spleen sections (paraffin-embedded) and am seeing a lot of autofluorescence, which I presume is due to platelets. Is there any way around this problem? Helen Newbery, Medical Genetics Section, Molecular Medicine Centre, Western General Hospital, Edinburgh, EH4 2XU. 0131 651 1047 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 DOOLEEO <@t> shands.ufl.edu Tue May 25 10:18:04 2004 From: DOOLEEO <@t> shands.ufl.edu (Elaine Dooley) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] mamoglobin Message-ID: Dear Histonetter, Vendors,.... Does anyone know of a commercial source for an anti-mamoglobin antibody that works on formalin fixed paraffin embedded material? Does it need a pretreatment? Thanks in advance Elaine Dooley HTL Shands Teaching Hospital Gainesville FL 352-265-0111 ext 72117 From mward <@t> wfubmc.edu Tue May 25 10:33:35 2004 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] alpha-HCG antibody Message-ID: <61135F0455D33347B5AAE209B903A304076A4E7E@EXCHVS2.medctr.ad.wfubmc.edu> I am looking for a vendor for alpha-HCG. We used to get it from Zymed and they no longer carry it. I have looked through the usual catalogs but keep coming up empty. Thanks in advance for your help. Martha Ward Wake Forest University Baptist Medical Center From Stacy_McLaughlin <@t> cooley-dickinson.org Tue May 25 10:43:36 2004 From: Stacy_McLaughlin <@t> cooley-dickinson.org (Stacy McLaughlin) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Grosslab 1" adhesive ruler strips Message-ID: <3D502BBF5356D31184650090275B750D0346C78B@mail.cooley-dickinson.org> Hello out there in Histoland. I hope someone can help me with this. I am looking for a 1" wide adhesive ruler strip that will stick to the front portion of the grossing station. It's marked up to 100cm increments. Thermo Electron (formerly Shandon) does not manufacture these anymore. Does anyone know of where I could get these? Thanks in advance for your help. Stacy THIS E-MAIL MESSAGE 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 the reader of this e-mail message is not the intended recipient, or the employee or agent responsible for delivering the message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this e-mail message is strictly prohibited. If you have received this e-mail message in error, please immediately notify Cooley Dickinson Healthcare at 413-582-2000 and delete or shred the original message and all copies thereof. Thank you. From amos <@t> dressi.com Tue May 25 10:50:51 2004 From: amos <@t> dressi.com (Amy R.) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Steven Slapp Message-ID: <7B284BA8386B394DA05712C1DFD0C7AB07228F@01server.diversifiedrealestate.local> Best wishes Steven. My father had a heart attack at 50. The doc said 3/4 of his heart dried up like a sponge & he wouldn't live another 24 hours. We had 4 more years with my dad---Anything's possible. We'll keep sending prayers. Amy Senn Managing Director of Appraisal Services Diversified National Service Center 100 West Mall Plaza, Carnegie PA 15106 (p) 888-345-6867 ext 262 (f) 412-446-0020 (e) amos@dressi.com Yahoo: diversifiednsc "Energy and persistence conquer all things." ~ Benjamin Franklin This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From GDawson <@t> Milw.Dynacare.com Tue May 25 10:59:26 2004 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] IHC Cocktails: Can we charge multiples? Message-ID: All, I recently attended a conference where I was informed that we can charge for mulitple antibodies when we do an IHC cocktail. For instance; my melanoma cocktail has 3 different antibodies in it and we've always just charged one 88342 for it, but could we charge 88342 X 3 for each of these that we run? I'm just wondering if anyone out there does charge for each antibody in their IHC cocktails and, if so, whether or not any problems arose from the practice. Also, are there any references that say in black and white that this is all on the up & up? Thanx In Advance, Glen Dawson Milwaukee, WI From juan.gutierrez <@t> christushealth.org Tue May 25 11:04:00 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] alpha-HCG antibody Message-ID: USBiolabs has one, though I've never used it. You might want to give them a ring. 1-800-520-3011 www.usbio.net -----Original Message----- From: Martha Ward [mailto:mward@wfubmc.edu] Sent: Tuesday, May 25, 2004 10:34 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] alpha-HCG antibody I am looking for a vendor for alpha-HCG. We used to get it from Zymed and they no longer carry it. I have looked through the usual catalogs but keep coming up empty. Thanks in advance for your help. Martha Ward Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mrsgbd2001 <@t> yahoo.com Tue May 25 11:11:02 2004 From: mrsgbd2001 <@t> yahoo.com (Gareth Davis) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Cost of Ventana Benchmark XT? In-Reply-To: <6EDD9DC298BF9C48AC3EF66EA92596BC14E398E4@frbucmsx03medge> Message-ID: <20040525161102.40421.qmail@web13308.mail.yahoo.com> Our lab bought two last year. The list price was $120,000 each. Gareth Davis PathGroup Nashville, Tennessee "Asarpota, Hitesh (MED, Intern)" wrote: Hello, Does someone know the approx cost of Cost of Ventana Benchmark XT in the U.S ? Tx, Hitesh Asarpota ---------------------------------------------------------- GE HealthCare Business Development Tel: +33 01 30 70 9224 Cell: +33 06 1171 4370 ---------------------------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? Friends. Fun. Try the all-new Yahoo! Messenger From JWEEMS <@t> sjha.org Tue May 25 10:53:31 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Steven Slapp Message-ID: Can anyone give update today? Thanks, j -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Amy R. Sent: Tuesday, May 25, 2004 11:51 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Steven Slapp Best wishes Steven. My father had a heart attack at 50. The doc said 3/4 of his heart dried up like a sponge & he wouldn't live another 24 hours. We had 4 more years with my dad---Anything's possible. We'll keep sending prayers. Amy Senn Managing Director of Appraisal Services Diversified National Service Center 100 West Mall Plaza, Carnegie PA 15106 (p) 888-345-6867 ext 262 (f) 412-446-0020 (e) amos@dressi.com Yahoo: diversifiednsc "Energy and persistence conquer all things." ~ Benjamin Franklin This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. _______________________________________________ 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 james.zimmerman <@t> pharma.novartis.com Tue May 25 10:41:25 2004 From: james.zimmerman <@t> pharma.novartis.com (james.zimmerman@pharma.novartis.com) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Antigen Message-ID: Hello, We're having trouble with tissue staying on the slide with EDTA antigen retrieval (both microwave and steam). Has anyone encountered this problem and if so how did you solve it ? Thanks, JPZ From ploykasek <@t> phenopath.com Tue May 25 11:07:42 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] mamoglobin In-Reply-To: Message-ID: Mammaglobin is available from Zeta Corporation located in Sierra Madre, CA phone #626-355-2053. We use this antibody with a heat retrieval in citrate buffer. Patti Loykasek PhenoPath Laboratories Seattle, WA > Dear Histonetter, Vendors,.... > > Does anyone know of a commercial source for an anti-mamoglobin antibody > that works on formalin fixed paraffin embedded material? Does it need a > pretreatment? > > Thanks in advance > Elaine Dooley HTL > Shands Teaching Hospital > Gainesville FL > 352-265-0111 ext 72117 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kzhong888 <@t> yahoo.com Tue May 25 11:53:52 2004 From: kzhong888 <@t> yahoo.com (dfs dsaf) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] NEw to histonet Message-ID: <20040525165352.53336.qmail@web41609.mail.yahoo.com> Hi all: i am a mohs tech new to histo net. i was wondering if any one has any old manuals for IEC cryostats or even old IEC cryostats for sale. Thanks Kirk Zhong, Vincent C Hung MD. INC. --------------------------------- Do you Yahoo!? Friends. Fun. Try the all-new Yahoo! Messenger From Rcartun <@t> harthosp.org Tue May 25 12:26:43 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] IHC Cocktails: Can we charge multiples? Message-ID: What antibody cocktails are you referring to? I would think that in order to charge multiple antibodies in a "cocktail" you would need to be able to discriminate the patterns of immunoreactivity for each antibody. For example, you should not be able to charge for two antibodies performed on the same slide when they both yield cytoplasmic immunoreactivity unless, of course, you use two different chromogens. Personally, we should be doing everything within our power to reduce the cost of health care; not make it more expensive. Richard Cartun >>> "Dawson, Glen" 05/25/04 11:59AM >>> All, I recently attended a conference where I was informed that we can charge for mulitple antibodies when we do an IHC cocktail. For instance; my melanoma cocktail has 3 different antibodies in it and we've always just charged one 88342 for it, but could we charge 88342 X 3 for each of these that we run? I'm just wondering if anyone out there does charge for each antibody in their IHC cocktails and, if so, whether or not any problems arose from the practice. Also, are there any references that say in black and white that this is all on the up & up? Thanx In Advance, Glen Dawson Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue May 25 12:46:10 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Autofluorescence problems in spleen Message-ID: <3.0.6.32.20040525114610.00c04170@gemini.msu.montana.edu> Since autofluorescence problems tend to be light greenish/yellowish tints, the problem is usually fainter than a good strong signal from Alexa 488 and sometimes fluorescein. One can use a brighter fluorophore (Alexa 488) or better yet, use the autofluorescence to your advantage and use a contrasting color fluorophore e.b. Rhodamine X (Jackson ImmunoResearch) TRITC-aka another rhodamine, Texas Red, or Alexa 546. If you are using eGFP, then you will have problems, unless the autofluorescence masks the GFP locale. In that case a good antiGFP antibody, then come back with a fluorophore that brightens up what you want to see. Formalin does create more autofluorescence but if you KNOW what is autofluorescing, you can sort it out better. There is a wonderful review on autofluorescence that I have in PDF file, but I cannot attach that to Histonet. If you contact me privately, I will send to you. The publication deals with GFP, fixatives and autofluorescence, but still applies to immunofluorescent staining. Also, go to Clontech website and look up the manual titled Living Colours, also for GFP, but this tidy little publication will tell you more details about what causes problems and how some people deal with it when working with GFP (Green Fluorescent Protein). People have attempted to get rid of autofluorescence, but we have far less problems with frozen sections to avoid formalin or paraformaldehyde fixation (the latter two are prefered fixatives for GFP) although the problem still exists to some extent with acetone or acetone/alcohol fixed frozen sections. 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 LESCHUKC <@t> trinity-health.org Tue May 25 13:36:35 2004 From: LESCHUKC <@t> trinity-health.org (Carmen Leschuk) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] NIST thermometers Message-ID: Hello, I purchased NIST spirit-filled thermometers a few years ago and recently pulled the certification that I received from them and noticed that it says it is only valid for one year! In lieu of the new CAP question ANP.23090, I am wondering if I need to purchase a different type of NIST thermometer. Any suggestions? Also, does anyone have a procedure on QC of their thermometers? Thank you! Carmen Leschuk, HT (ASCP) Supervisor, SJMO-Anatomic Pathology (248)858-6231 From Jackie.O'Connor <@t> abbott.com Tue May 25 13:47:09 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] NIST thermometers Message-ID: Our calibration department regularly calibrates thermometers, pipets, equipment (i.e., waterbaths, refrigerators) to ensure they are reporting accurate temperatures and measurements. Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotherapeutics "Carmen Leschuk" Sent by: histonet-bounces@lists.utsouthwestern.edu 05/25/2004 01:36 PM To: histonet@lists.utsouthwestern.edu, histonet-request@lists.utsouthwestern.edu cc: Subject: [Histonet] NIST thermometers Hello, I purchased NIST spirit-filled thermometers a few years ago and recently pulled the certification that I received from them and noticed that it says it is only valid for one year! In lieu of the new CAP question ANP.23090, I am wondering if I need to purchase a different type of NIST thermometer. Any suggestions? Also, does anyone have a procedure on QC of their thermometers? Thank you! Carmen Leschuk, HT (ASCP) Supervisor, SJMO-Anatomic Pathology (248)858-6231 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mprice26 <@t> juno.com Tue May 25 14:06:56 2004 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Osmium Tetroxide Protocol Message-ID: <20040525.120714.16726.81860@webmail28.nyc.untd.com> Hi Histonetters, When handling Osmium Tetroxide, is a chemical fume gard such as an Bench Top fume absorber appropriate to use for this chemical? Also, should you wear other PPE besides gloves, gown etc. Should you wear a mask also? Thank you. Marsha Price ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! From carl.hobbs <@t> kcl.ac.uk Tue May 25 14:28:29 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] re antigen Message-ID: <004201c4428e$7544aa00$7dcc9a51@home> same problem with EDTA but what's the pH of your solution??....I use either citric acid at pH6 or TRIS at pH10. Get excellent results, tho' citric is not so kind to my tissues, usually. Don't understand it.........got to try all pHs and the posted range of solutions to find your own way ( two different labs can have opposite results with same solution...right, Stan???) --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.690 / Virus Database: 451 - Release Date: 22/05/2004 From Stacy_McLaughlin <@t> cooley-dickinson.org Tue May 25 14:36:45 2004 From: Stacy_McLaughlin <@t> cooley-dickinson.org (Stacy McLaughlin) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Osmium Tetroxide Protocol Message-ID: <3D502BBF5356D31184650090275B750D0346C78F@mail.cooley-dickinson.org> Marsha, When in doubt, I look at the product's MSDS. It should have all the info you need for safe handling of the chemical. Stacy -----Original Message----- From: mprice26@juno.com [mailto:mprice26@juno.com] Sent: Tuesday, May 25, 2004 2:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Osmium Tetroxide Protocol Hi Histonetters, When handling Osmium Tetroxide, is a chemical fume gard such as an Bench Top fume absorber appropriate to use for this chemical? Also, should you wear other PPE besides gloves, gown etc. Should you wear a mask also? Thank you. Marsha Price ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet THIS E-MAIL MESSAGE 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 the reader of this e-mail message is not the intended recipient, or the employee or agent responsible for delivering the message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this e-mail message is strictly prohibited. If you have received this e-mail message in error, please immediately notify Cooley Dickinson Healthcare at 413-582-2000 and delete or shred the original message and all copies thereof. Thank you. From Stanley.Stylli <@t> mh.org.au Tue May 25 14:37:40 2004 From: Stanley.Stylli <@t> mh.org.au (Stylli, Stanley) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] re antigen Message-ID: <00519F9F41D2844D9C75BEE1F6AC09AF01992D29@rmhmail1.ssg.org.au> carl...you took the words right out of my mouth ! I totally agree..... nothing is more frustrating when you are using the identical protocol as someone else and you get the opposite results ! (must be the water down under !) -----Original Message----- From: Carl [mailto:carl.hobbs@kcl.ac.uk] Sent: Wednesday, 26 May 2004 5:28 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] re antigen same problem with EDTA but what's the pH of your solution??....I use either citric acid at pH6 or TRIS at pH10. Get excellent results, tho' citric is not so kind to my tissues, usually. Don't understand it.........got to try all pHs and the posted range of solutions to find your own way ( two different labs can have opposite results with same solution...right, Stan???) --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.690 / Virus Database: 451 - Release Date: 22/05/2004 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lesley <@t> vancouverbc.net Tue May 25 14:57:18 2004 From: lesley <@t> vancouverbc.net (Lesley Weston) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Osmium Tetroxide Protocol In-Reply-To: <20040525.120714.16726.81860@webmail28.nyc.untd.com> Message-ID: It's better than nothing, but not really adequate. OsO4 is really nasty stuff; you should treat it with great caution. A mask won't help at all, unless you mean a full-scale respirator - it's fumes you're worried about, not particles - but you should certainly wear gloves and a lab-coat, and change the gloves frequently. If you can't use a real fume hood, properly-fitting goggles might be an idea, too; the fumes can fix the cornea permanently, and possibly even affect the retina. Do you have the MSDS sheet for OsO4? That would give you all the information you need. Lesley Weston. on 25/05/2004 12:06 PM, mprice26@juno.com at mprice26@juno.com wrote: > > Hi Histonetters, > > When handling Osmium Tetroxide, is a chemical fume gard such as an Bench Top > fume absorber appropriate to use for this chemical? > > Also, should you wear other PPE besides gloves, gown etc. Should you wear a > mask also? > > Thank you. > > Marsha Price > > ________________________________________________________________ > The best thing to hit the Internet in years - Juno SpeedBand! > Surf the Web up to FIVE TIMES FASTER! > Only $14.95/ month - visit www.juno.com to sign up today! > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Tue May 25 15:06:45 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Osmium Tetroxide Protocol Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA206EB58@usca0082k08.labvision.apogent.com> Here's a rule of thumb for osmium: if you can smell it, it is fixing you! (from experience - it smells a bit like chlorine). And if yo can smell it, your eyes are in danger - that is the primary concern for osmium. You are much better off using a true fume hood. Tim Morken Lab Vision - Neomarkers www.labvision.com -----Original Message----- From: mprice26@juno.com [mailto:mprice26@juno.com] Sent: Tuesday, May 25, 2004 12:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Osmium Tetroxide Protocol Hi Histonetters, When handling Osmium Tetroxide, is a chemical fume gard such as an Bench Top fume absorber appropriate to use for this chemical? Also, should you wear other PPE besides gloves, gown etc. Should you wear a mask also? Thank you. Marsha Price ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> colobio.com Tue May 25 15:18:42 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] IHC Cocktails: Can we charge multiples? In-Reply-To: Message-ID: I agree with Rich on this, why should you charge extra when you have not done extra work and the antibody cocktails don't usually cost anymore than single antibodies, actually when you think about it these cocktail procedures should probably be cheaper for the patient not more. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Richard Cartun Sent: Tuesday, May 25, 2004 11:27 AM To: histonet@lists.utsouthwestern.edu; GDawson@Milw.Dynacare.com Subject: Re: [Histonet] IHC Cocktails: Can we charge multiples? What antibody cocktails are you referring to? I would think that in order to charge multiple antibodies in a "cocktail" you would need to be able to discriminate the patterns of immunoreactivity for each antibody. For example, you should not be able to charge for two antibodies performed on the same slide when they both yield cytoplasmic immunoreactivity unless, of course, you use two different chromogens. Personally, we should be doing everything within our power to reduce the cost of health care; not make it more expensive. Richard Cartun >>> "Dawson, Glen" 05/25/04 11:59AM >>> All, I recently attended a conference where I was informed that we can charge for mulitple antibodies when we do an IHC cocktail. For instance; my melanoma cocktail has 3 different antibodies in it and we've always just charged one 88342 for it, but could we charge 88342 X 3 for each of these that we run? I'm just wondering if anyone out there does charge for each antibody in their IHC cocktails and, if so, whether or not any problems arose from the practice. Also, are there any references that say in black and white that this is all on the up & up? Thanx In Advance, Glen Dawson Milwaukee, WI _______________________________________________ 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 mrsgbd2001 <@t> yahoo.com Tue May 25 16:10:04 2004 From: mrsgbd2001 <@t> yahoo.com (Gareth Davis) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Good cryosections of rat retina? Message-ID: <20040525211004.13100.qmail@web13306.mail.yahoo.com> Hello all, This week I am trying to get good cryosections of rat eyes. We are mostly interested in the retinas. The eyes I have were from animals perfused with 4% Paraformaldehyde, eyes were post-fixed with same, then cryopreserved. Eyes were placed in OCT for cutting. The eyes were not cut in half before placing in OCT. I am having a little trouble getting good sections. I would just like to get some ideals of how others prepare animal eyes for sectioning. Thanks, Gareth Davis Research Assistant Tennessee State University Nashville, Tennessee --------------------------------- Do you Yahoo!? Friends. Fun. Try the all-new Yahoo! Messenger From ploykasek <@t> phenopath.com Tue May 25 16:30:48 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:23:01 2005 Subject: FW: [Histonet] IHC Cocktails: Can we charge multiples? In-Reply-To: Message-ID: ------ Forwarded Message From: Patti Loykasek Date: Tue, 25 May 2004 09:18:29 -0700 To: "Dawson, Glen" Subject: Re: [Histonet] IHC Cocktails: Can we charge multiples? Glen, it has been my understanding that you can charge for the antibodies in a cocktail if you can detect each antibody separately. In other words, if the staining pattern of the antibodies is different. We have used a melanoma cocktail in the past, but no longer do this for several reasons. All of the antibodies in the cocktail gave the same cytoplasmic staining pattern, and we only charged for one. Also, if 1 antibody was not performing optimally you would never know it since all have the same staining pattern. I certainly am no charging/compliance guru, just my thoughts on the subject. Patti Loykasek PhenoPath Laboratories Seattle, WA> > All, > > I recently attended a conference where I was informed that we can charge for > mulitple antibodies when we do an IHC cocktail. For instance; my melanoma > cocktail has 3 different antibodies in it and we've always just charged one > 88342 for it, but could we charge 88342 X 3 for each of these that we run? > > I'm just wondering if anyone out there does charge for each antibody in > their IHC cocktails and, if so, whether or not any problems arose from the > practice. Also, are there any references that say in black and white that > this is all on the up & up? > > Thanx In Advance, > > Glen Dawson > Milwaukee, WI > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------ End of Forwarded Message From peptolab <@t> hamptons.com Tue May 25 18:17:37 2004 From: peptolab <@t> hamptons.com (peptolab) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] Price gouging on cocktails?? Message-ID: <001f01c442ae$7906fba0$687bbd18@JEFF> Glen- If your cocktail is positive, can you say which of the three melanoma markers (I presume HMB 45, Mart-1, and tyrosinase) are responsible for the positivity? If no, then maybe only one charge is appropriate as this is a screening test and you are only screening one slide. Does the antibody cocktail cost three times the cost of any single component marker? Are you running controls for all three markers separately or just a control for the cocktail. Maybe one component isn't staining well- hw would you know? I vote for one charge. Jeff Silverman HT HTL QIHC (ASCP) From yichaowu <@t> hotmail.com Tue May 25 19:50:59 2004 From: yichaowu <@t> hotmail.com (yichao wu) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] ROOM TEMPERATURE RE: C4d antibody Message-ID: Dear Gareth, Thank you for your attention. Yes I agree the monoclonal antibody A213 may be not appliable on paraffin-embedded sections without pretreatment. I immerse the sections in 88% formic acid/ddH2O for 20 minutes just at room temperature. I think the extreme acid environment would help to uncover the antigen. And I also tried other pretreatments such as digestion by 0.1% typsin, heat pretreatment in citric acid and so on.But the signal did not shown up. However, I should mention that the fixation solution is also very important besides the paraffin embedding. I wonder what kind of fixation solution you use. Actually ours is a little different from normal 10% formalin even if I suppose that sections fixed by 10% buffered formalin may be tried with formic acid as well. Besides, I use FITC-conjugated Rabbit-anti-mouse IgG as the secondary antibody later in my experiment.And the fluorescence signals seem more easier to be observed by the eye. As a postgraduate student myself, I am not very experienced in the field of immunohistology. However, I have get warm help from histonet. And I do wish to communicate with more histologists here. Sincerely, Yichao Yichao WU, MD Department of Medicine Nanjing University School of Medicine Nanjing, China Email: yichaowu@hotmail.com TEL: +86-25-80860805 >From: "Gareth Bruce" >To: >Subject: C4d antibody >Date: Tue, 25 May 2004 16:25:57 +0100 > >Dear Yichao WU, > > > >I noticed on histonet that you have experience with Quidel's antibody A213 >to C4d. > > > >We have been experiencing some difficulty using this antibody in IHC with >paraffin sections. > >You recommend 20 minutes of 88% formic acid pretreatment. > >May I ask you what temperature this was done at? > > > >Thank you so very much. > > > >Best regards > > > >Gareth Bruce > > > _________________________________________________________________ Add photos to your e-mail with MSN 8. Get 2 months FREE*. http://join.msn.com/?page=features/featuredemail From bills <@t> icpmr.wsahs.nsw.gov.au Wed May 26 00:58:38 2004 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:23:01 2005 Subject: [Histonet] BK virus detection. Message-ID: <000001c442e6$7e280600$e287080a@wsahs.nsw.gov.au> Dear Histonetters, I am writing for a colleague who performs all our Immunohistochemistry. She is asking: "Can anyone help us with a reliable method of the detection of BK virus in Renal Transplant biopsies. We use a Chemicon anti-BK virus and treat the sections in citrate buffer in a pressure cooker in the microwave. A polymer based peroxidase kit is used for detection. Unfortunately this method gives us a great deal of non-specific staining and almost every nucleus of every cell in the tissue is stained, no what the dilution of the antibody. We would be grateful if anyone can impart the secret of their success with us. Many thanks Jane" Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 __________________________________________________________________ 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 marshall <@t> cormack.uct.ac.za Wed May 26 05:50:33 2004 From: marshall <@t> cormack.uct.ac.za (Marshall) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] exotic species/chief&parietal cells Message-ID: <40B47679.9622A362@cormack.uct.ac.za> Hi all We are looking at the gastric mucosa of the Cape Mole Rat and can not distinguish chief cells on the basis of morphology. There seem to be more parietal cells and we can not be sure of the chief cells. Is there a stain we can use to distinguish between these to cell types as the Hematoxylin & Eosin stain is not making this obvious. Thanks Sharon Marshall Dept. Human Biology University of Cape Town South Africa e-mail: marshall@cormack.uct.ac.za From ASelf <@t> gmhsc.com Wed May 26 06:33:36 2004 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] H&E automatic slide stainer Message-ID: <39836CD6DB61654E8F95A35898C921860A86FD@exchange.gmhpost.com> Hello everyone, We are looking to budget an H&E automatic slide stainer..... we are a small histology lab with limited space and just two of us working. We are looking for something that is small/compact but at the same time within our budget. Does anyone have any recommendations or any suggestions with this. We have come across one stainer from Richard Alan that is within our spending(less than $15,000). It is a compact linear stainer - does anyone have any experience with this type of stainer. If so how do you like it. Thanks for your help, Amy NOTE: The information contained in this message may be privileged, 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 this message and deleting it from your computer. Thank you. From jqb7 <@t> cdc.gov Wed May 26 06:40:53 2004 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] H&E automatic slide stainer Message-ID: Many years ago I had a linear stainer and had no problems with it. I would suggest that you see if Richard-Allan will let you demo it. That will give you the best shot to see if it fits your needs. Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Self Sent: Wednesday, May 26, 2004 7:34 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] H&E automatic slide stainer Hello everyone, We are looking to budget an H&E automatic slide stainer..... we are a small histology lab with limited space and just two of us working. We are looking for something that is small/compact but at the same time within our budget. Does anyone have any recommendations or any suggestions with this. We have come across one stainer from Richard Alan that is within our spending(less than $15,000). It is a compact linear stainer - does anyone have any experience with this type of stainer. If so how do you like it. Thanks for your help, Amy NOTE: The information contained in this message may be privileged, 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 this message and deleting it from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vandries <@t> vub.ac.be Wed May 26 07:03:01 2004 From: vandries <@t> vub.ac.be (Veronique Andriessen) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] ki67 Message-ID: Hello histonetters, I'm searching for a Ki67 antibody that will work on frozen sections of rat pancreas. Can you give me some advice? Best regards, Veronique Andriessen BAS Lab. Molecular Liver Cell Biology, Free University Brussels (VUB) Belgium From Sarah.Blachford <@t> ngh.nhs.uk Wed May 26 08:05:09 2004 From: Sarah.Blachford <@t> ngh.nhs.uk (Blachford, Sarah - Histopath Main) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] (no subject) Message-ID: <854FA06D8BE2D611ABA30030052D1DF6FA58ED@NGH_EXCHANGE1> When performing immunofluorescence can you then add a substrate or any thing else to turn this into a non fluorosecent slide which can then be visible down a light microscope. S Northampton General Hospital NHS Trust Cliftonville, Northampton NN1 5BD This e-mail may contain confidential information and/or copyright material and is intended for the use of the addressee only. Any unauthorised use may be unlawful. The contents of this e-mail may be subject to public disclosure under the NHS Code of Openness or the Freedom of Information Act 2000. Unless legally exempt, the confidentiality of the message and your reply cannot be guaranteed. If you receive this e-mail by mistake, please advise the sender immediately. Thank you. From la.sebree <@t> hosp.wisc.edu Wed May 26 08:01:54 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] IHC Cocktails: Can we charge multiples? Message-ID: We charge one 88342 for all our antibody cocktails. 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 -----Original Message----- From: Dawson, Glen [mailto:GDawson@Milw.Dynacare.com] Sent: Tuesday, May 25, 2004 10:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC Cocktails: Can we charge multiples? All, I recently attended a conference where I was informed that we can charge for mulitple antibodies when we do an IHC cocktail. For instance; my melanoma cocktail has 3 different antibodies in it and we've always just charged one 88342 for it, but could we charge 88342 X 3 for each of these that we run? I'm just wondering if anyone out there does charge for each antibody in their IHC cocktails and, if so, whether or not any problems arose from the practice. Also, are there any references that say in black and white that this is all on the up & up? Thanx In Advance, Glen Dawson Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Wed May 26 08:10:20 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Good cryosections of rat retina? Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3B3B@exchsrv01.barrynet.barry.edu> One of my former students, James Gentile (now a prosthodontist in Philadelphia), got excellent results by using a fine gauge hypodermic needle to withdraw some of the vitreous humor and injecting Bouin's fluid into the vitreous chamber. He then immersed the eye in Bouin's for a few days. You might get similar results by injecting 4% paraformaldehyde into the vitreous chamber. 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 Gareth Davis Sent: Tuesday, May 25, 2004 5:10 PM To: Histonet Subject: [Histonet] Good cryosections of rat retina? Hello all, This week I am trying to get good cryosections of rat eyes. We are mostly interested in the retinas. The eyes I have were from animals perfused with 4% Paraformaldehyde, eyes were post-fixed with same, then cryopreserved. Eyes were placed in OCT for cutting. The eyes were not cut in half before placing in OCT. I am having a little trouble getting good sections. I would just like to get some ideals of how others prepare animal eyes for sectioning. Thanks, Gareth Davis Research Assistant Tennessee State University Nashville, Tennessee --------------------------------- Do you Yahoo!? Friends. Fun. Try the all-new Yahoo! Messenger _______________________________________________ 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 asmith <@t> mail.barry.edu Wed May 26 08:35:19 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Osmium Tetroxide Protocol Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3B3C@exchsrv01.barrynet.barry.edu> Osmium tetroxide is a gas, usually sold dissolved in water. It has a nasty tendency to evaporate from the solution. It reacts instantly with amino alcohols (cell membranes) and with cytokeratins (skin and cornea) to leave black deposits of osmium metal. Deposits in the cornea interfere with vision for many weeks. You must use it in a fume hood that draws really well. In one of my postdoctoral positions we didn't trust our fume hood and used osmium tetroxide outdoors only on days when the wind speed was 15 mph or greater. One, of course, was careful to stay upwind of one's work! Goggles or wrap-around safety glasses are also essential in case the #@$* stuff spatters. 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 mprice26@juno.com Sent: Tuesday, May 25, 2004 3:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Osmium Tetroxide Protocol Hi Histonetters, When handling Osmium Tetroxide, is a chemical fume gard such as an Bench Top fume absorber appropriate to use for this chemical? Also, should you wear other PPE besides gloves, gown etc. Should you wear a mask also? Thank you. Marsha Price ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! _______________________________________________ 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 asmith <@t> mail.barry.edu Wed May 26 08:42:33 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] exotic species/chief&parietal cells Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3B3D@exchsrv01.barrynet.barry.edu> Bowie's pepsin stain usually stains chief cells very well. D.J. Bowie (1936) Method for studying pepsinogen granules in gastric glands. Anatomical Record 64: 357-367. 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 Marshall Sent: Wednesday, May 26, 2004 6:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] exotic species/chief&parietal cells Hi all We are looking at the gastric mucosa of the Cape Mole Rat and can not distinguish chief cells on the basis of morphology. There seem to be more parietal cells and we can not be sure of the chief cells. Is there a stain we can use to distinguish between these to cell types as the Hematoxylin & Eosin stain is not making this obvious. Thanks Sharon Marshall Dept. Human Biology University of Cape Town South Africa e-mail: marshall@cormack.uct.ac.za _______________________________________________ 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 GDawson <@t> Milw.Dynacare.com Wed May 26 08:33:11 2004 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] RE: Price gouging on cocktails?? Message-ID: Wow, A legitimate question asked in an appropriate list-server leading to accusations of price gouging. I should have left the actual question out of my original post and just took a poll on how everyone feels about the out of control cost of health-care because that seems to be the more interesting topic. Moral/Ethical issues aside, I truly wanted to hear of any experience histo-land had with this issue because I actually did hear it at a conference (the presenter's name will be withheld to protect him from accusations of holding patients down and rifling through their pockets) and it actually did come up in my lab. Now I remember why I switched over to just reading the histonet and not daring to post anything in such a politically correct environment. I'll contact the workshop presenter to get the low-down on this question. Believe it or not, my intentions were not to price gouge patients that are already being raked over the coals by the health-care system in this country. IMHO; we should look at other issues such as the need to hire whole fleets of personelle just to wade through the hundreds of insurance companies and their accompanying issues. The histo lab is, as a rule, putting out a very reasonably priced service with profit margins on tests being kept low to out compete the rival lab that may try to take your business. If enough expensive antibodies are added to a cocktail, it can become less than profitable. I realize that the 88342 charge carries some exhorbitant professional fees along with it but perhaps a lab out there has been able to somehow raise just the technical fee a bit on cocktails??? Any insights on this topic would be much appreciated. More "how dare you's" would not. Thank-you, Glen Dawson -----Original Message----- From: peptolab [mailto:peptolab@hamptons.com] Sent: Tuesday, May 25, 2004 5:18 PM To: GDawson@Milw.Dynacare.com Cc: HistoNet Server Subject: Price gouging on cocktails?? Glen- If your cocktail is positive, can you say which of the three melanoma markers (I presume HMB 45, Mart-1, and tyrosinase) are responsible for the positivity? If no, then maybe only one charge is appropriate as this is a screening test and you are only screening one slide. Does the antibody cocktail cost three times the cost of any single component marker? Are you running controls for all three markers separately or just a control for the cocktail. Maybe one component isn't staining well- hw would you know? I vote for one charge. Jeff Silverman HT HTL QIHC (ASCP) From pwg1 <@t> cdc.gov Wed May 26 09:00:25 2004 From: pwg1 <@t> cdc.gov (Greer, Patricia) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] (no subject) Message-ID: Sarah, We sometimes use FITC labeled antibodies as our primary antibody, followed by either a mouse or rabbit anti-FITC then continue with your usual immuno procedure - in our case we use the LSAB2 kit from Dako. Pat Greer Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road Atlanta, GA 30333 When performing immunofluorescence can you then add a substrate or any thing else to turn this into a non fluorosecent slide which can then be visible down a light microscope. S Northampton General Hospital NHS Trust Cliftonville, Northampton NN1 5BD From gcallis <@t> montana.edu Wed May 26 09:23:03 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Post immunofluorescence immunostaining In-Reply-To: <854FA06D8BE2D611ABA30030052D1DF6FA58ED@NGH_EXCHANGE1> Message-ID: <3.0.6.32.20040526082303.00c0ecd0@gemini.msu.montana.edu> I suppose one could pull the coverslip at risk of damaging section and then come back with antiPE, antiFITC, or go to Molecular Probes and see if they have an antiAlexa 488 antibody. There are some nice antibodies for detecting fluorophores out there. I would suggest coverslipping with PBS for viewing with fluorescent microscope and do this immediately, without using any coverslip edge sealant. Soak off coverslip and try immunostaining. DAKO has an excellent Rabbit antiFITC, (do not use the rabbit antiFITC-HRP conjugate, it doesn't work as well per Chris van der Loos suggestion/experience) then come back with an antiRabbit secondary (Donkey antirabbit-biotin) etc. Just make sure you are careful with blocking and secondary hosts to prevent nonspecific cross reaction detection. At 03:05 PM 5/26/2004 +0200, you wrote: > >When performing immunofluorescence can you then add a substrate or any thing >else to turn this into a non fluorosecent slide which can then be visible >down a light microscope. > >S > > >Northampton General Hospital NHS Trust >Cliftonville, Northampton NN1 5BD > >This e-mail may contain confidential information and/or copyright material >and is intended for the use of the addressee only. Any unauthorised use >may be unlawful. The contents of this e-mail may be subject to public >disclosure under the NHS Code of Openness or the Freedom of Information Act >2000. Unless legally exempt, the confidentiality of the message and your >reply cannot be guaranteed. If you receive this e-mail by mistake, please >advise the sender immediately. >Thank you. > >_______________________________________________ >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 ian.montgomery <@t> bio.gla.ac.uk Wed May 26 09:36:57 2004 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:23:02 2005 Subject: Fwd: [Histonet] exotic species/chief&parietal cells Message-ID: <6.1.0.6.2.20040526152915.02d120d0@udcf.gla.ac.uk> Sharon, Haematoxylin, Eosin Phloxine and Tartrazine is a nice stain for these cells. Lendrum, AC (1947) J. Path. & Bact. 40. 399-404. If you cannot get the paper let me know and I'll e-mail the method. Ian. >Hi all >We are looking at the gastric mucosa of the Cape Mole Rat and can not >distinguish chief cells on the basis of morphology. There seem to be >more parietal cells and we can not be sure of the chief cells. Is there >a stain we can use to distinguish between these to cell types as the >Hematoxylin & Eosin stain is not making this obvious. >Thanks >Sharon Marshall >Dept. Human Biology >University of Cape Town >South Africa >e-mail: marshall@cormack.uct.ac.za > >_______________________________________________ >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 lizchlipala <@t> premierhistology.com Wed May 26 10:22:19 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] ki67 In-Reply-To: Message-ID: <000001c44335$3f97fef0$74d48a80@LIZ> Veronique We use a Rat Ki-67 from Dakocytomation (M7248) we use this on formalin fixed paraffin embedded specimens with steam retrieval. I'm assuming it would work on frozen sections. I have a protocol and some images if you would like me to send them to you. Thanks Liz Elizabeth A. Chlipala, BS, HTL(ASCP) 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 MCBD, Room A3B40 Boulder, Colorado 80309 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Veronique Andriessen Sent: Wednesday, May 26, 2004 5:03 AM To: Histonet Subject: [Histonet] ki67 Hello histonetters, I'm searching for a Ki67 antibody that will work on frozen sections of rat pancreas. Can you give me some advice? Best regards, Veronique Andriessen BAS Lab. Molecular Liver Cell Biology, Free University Brussels (VUB) Belgium _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mhorne <@t> upei.ca Wed May 26 10:57:26 2004 From: mhorne <@t> upei.ca (Margaret Horne) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Osmium Tetroxide Protocol In-Reply-To: <20040525.120714.16726.81860@webmail28.nyc.untd.com> Message-ID: <40B49436.28709.1006818@localhost> Hi , when I handle osmium I always : a) Carry the Osmium to the fume hood in a plastic container . ie. the Osmium solution is in a glass jar with a ground glass stopper and heavily parafilmed. Also the jar is covered in aluminum foil ( to avoid degradation from the light). This jar is Always carried about in a plastic container with a lid. Heavy rubbermaid or tupperware is good. Note that you will see the plastic container turn black over time with the Osmium fumes escaping. We keep it in the " Dirty " fridge , not the " Clean" fridge with our antibodies. b) The plastic container is opened in the fume hood and I wear a labcoat and am Doubled Gloved. If the outer glove shows areas of black , then the glove integrety has been compromised and I replace the outer glove. Double gloving can be uncomfortably tight. I found that a nitrile for the inner glove and a latex for the outer glove is comfortable for me but it depends on the fit of the gloves you use . In a pinch I have used two pair of latex. c) Any glasswear that I use , I leave in the fume hood overnight for the fumes to evaporate. d) I keep a container of oil in the fume hood when i am working with the osmium so I can pour oil on a spill if necesary or to neutralise the osmium if mixed with , say , blood , because it binds to lipids preferentially. I have used vegetable oil but it goes rancid over time. Mineral oil is fine. Remember that osmium fumes alone are sometimes used to fix tissue. So working without proper ventilation means your eyeballs are going to become fixed. Insist on a proper fume hood. Other than that , it is not bad stuff to work with :-) Margaret ps. remember that osmium penetrates only 1 mm in depth so your tissue has to be diced small. Margaret Horne , Histology Teaching Assistant, Dept. of B.SC., Atlantic Veterinary College, U.P.E.I., 550 University Ave., Charlottetown, P.E.I., C1A 4P3 Canada From komo <@t> bu.edu Wed May 26 11:02:16 2004 From: komo <@t> bu.edu (Rob Komorowski) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Anyone know of a good cartoon showing a basic immuno protocol? In-Reply-To: <200405051558.i45FwOBf018057@relay9.bu.edu> Message-ID: Hi all, I'm trying to find a good step by step cartoon showing a primary antibody linking up with an antigen, a biotinylated secondary antibody, a strepHRP step, and a DAB reaction. I know...sounds specific...but anything close would be helpful for teaching. Thanks! Rob K From tpmorken <@t> labvision.com Wed May 26 11:21:32 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Osmium Tetroxide Protocol and mineral oil? Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA206EB5C@usca0082k08.labvision.apogent.com> Margaret wrote: <> I have not ever seen a reference of mineral oil being used to "neutralize" osmium. Osmium reacts with unsaturated oils and mineral oil, as far as I know is fully saturated. In fact, I imagine that what happens is that the osmium is simply dissolved in the mineral oil (similar to how it dissolves in xylene). I really doubt the osmium is being neutralized by the mineral oil. The following references give the proper procdure for using corn oil (poly unsaturated) to neutralize osmium. http://www.cbwinfo.com/Chemical/HistandMisc/oso4.shtml http://www.wfubmc.edu/ehs/sop/osmimtw.pdf http://www.proscitech.com.au/catalogue/notes/c010.htm If you have a reference to support the mineral oil neutralization I'd certainly like to see it. Tim Morken -----Original Message----- From: Margaret Horne [mailto:mhorne@upei.ca] Sent: Wednesday, May 26, 2004 5:57 AM To: histonet@lists.utsouthwestern.edu; mprice26@juno.com Subject: Re: [Histonet] Osmium Tetroxide Protocol Hi , when I handle osmium I always : a) Carry the Osmium to the fume hood in a plastic container . ie. the Osmium solution is in a glass jar with a ground glass stopper and heavily parafilmed. Also the jar is covered in aluminum foil ( to avoid degradation from the light). This jar is Always carried about in a plastic container with a lid. Heavy rubbermaid or tupperware is good. Note that you will see the plastic container turn black over time with the Osmium fumes escaping. We keep it in the " Dirty " fridge , not the " Clean" fridge with our antibodies. b) The plastic container is opened in the fume hood and I wear a labcoat and am Doubled Gloved. If the outer glove shows areas of black , then the glove integrety has been compromised and I replace the outer glove. Double gloving can be uncomfortably tight. I found that a nitrile for the inner glove and a latex for the outer glove is comfortable for me but it depends on the fit of the gloves you use . In a pinch I have used two pair of latex. c) Any glasswear that I use , I leave in the fume hood overnight for the fumes to evaporate. d) I keep a container of oil in the fume hood when i am working with the osmium so I can pour oil on a spill if necesary or to neutralise the osmium if mixed with , say , blood , because it binds to lipids preferentially. I have used vegetable oil but it goes rancid over time. Mineral oil is fine. Remember that osmium fumes alone are sometimes used to fix tissue. So working without proper ventilation means your eyeballs are going to become fixed. Insist on a proper fume hood. Other than that , it is not bad stuff to work with :-) Margaret ps. remember that osmium penetrates only 1 mm in depth so your tissue has to be diced small. Margaret Horne , Histology Teaching Assistant, Dept. of B.SC., Atlantic Veterinary College, U.P.E.I., 550 University Ave., Charlottetown, P.E.I., C1A 4P3 Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed May 26 11:22:46 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Re:good cartoon showing a basic immunoprotocol In-Reply-To: References: <200405051558.i45FwOBf018057@relay9.bu.edu> Message-ID: <3.0.6.32.20040526102246.00c08980@gemini.msu.montana.edu> DAKO has these in their Handbook, also any good immunostaining textbook, but beware, if you are using their publications, you must ask permission to use to not violate copyright laws. Jackson Immunoresearch has cartoons also. You can be creative and draw your own, right into a Powerpoint presentation. Very easy to do with all the bells and whistles in "draw". That way you see have as a template but do not violate copyright. Your own creations are usually far nicer for colors, etc. Scanned pictures, cartoons from publications are are generally grainy anyway. When you do the lecture, you can add animation to make each step (primary detecting antigen, secondary detecting primary, etc be much more dynamic - have done it for my student lectures and workshops. I find a single flat picture handed to students during lecture is confusing, but if it is done step by step, they grasp concepts faster. A big help is to download the DAKO handbook as a pdf, burn to CD's and hand to students for their use in future. DAKO has sent me a dozen handbooks for class use in past, but CD was very nice and compact. > 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 zandra <@t> gwu.edu Wed May 26 11:51:36 2004 From: zandra <@t> gwu.edu (Alexandra de Sousa) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] SMI-32 troubles Message-ID: <16182c9161a83f.161a83f16182c9@gwu.edu> Hello. I have trying to use SMI-32 as a marker in primate visual cortex for some time, and my results have been consistantly poor. I get lots of background, and sparse, faintly stained neurons. My sections are mostly immersion fixed so I use an antigen retrieval. I have been using the Vector ABC method, has any one had better results with PAP or something else? I would really appreciate any advice from people who have done immuno on primates, or have figured out ways to improve SMI-32 results! Thanks! Alexandra de Sousa From gcallis <@t> montana.edu Wed May 26 11:53:41 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Osmium Tetroxide Protocol and mineral oil? In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA206EB5C@usca0082k08.labvisi on.apogent.com> Message-ID: <3.0.6.32.20040526105341.00c0d400@gemini.msu.montana.edu> I agree, corn oil was what I was told also. We also had our chemical safety people collect this waste. At 09:21 AM 5/26/2004 -0700, you wrote: >Margaret wrote: ><the osmium so I can pour oil on a spill if necesary or to neutralise >the osmium if mixed with , say , blood , because it binds to lipids >preferentially. I have used vegetable oil but it goes rancid over time. >Mineral oil is fine.>> > >I have not ever seen a reference of mineral oil being used to "neutralize" >osmium. Osmium reacts with unsaturated oils and mineral oil, as far as I >know is fully saturated. In fact, I imagine that what happens is that the >osmium is simply dissolved in the mineral oil (similar to how it dissolves >in xylene). I really doubt the osmium is being neutralized by the mineral >oil. The following references give the proper procdure for using corn oil >(poly unsaturated) to neutralize osmium. > >http://www.cbwinfo.com/Chemical/HistandMisc/oso4.shtml >http://www.wfubmc.edu/ehs/sop/osmimtw.pdf >http://www.proscitech.com.au/catalogue/notes/c010.htm > >If you have a reference to support the mineral oil neutralization I'd >certainly like to see it. > >Tim Morken > > > >-----Original Message----- >From: Margaret Horne [mailto:mhorne@upei.ca] >Sent: Wednesday, May 26, 2004 5:57 AM >To: histonet@lists.utsouthwestern.edu; mprice26@juno.com >Subject: Re: [Histonet] Osmium Tetroxide Protocol > > >Hi , when I handle osmium I always : > >a) Carry the Osmium to the fume hood in a plastic container . ie. >the Osmium solution is in a glass jar with a ground glass stopper >and heavily parafilmed. Also the jar is covered in aluminum foil ( to >avoid degradation from the light). This jar is Always carried about in >a plastic container with a lid. Heavy rubbermaid or tupperware is >good. Note that you will see the plastic container turn black over >time with the Osmium fumes escaping. We keep it in the " Dirty " >fridge , not the " Clean" fridge with our antibodies. > >b) The plastic container is opened in the fume hood and I wear a >labcoat and am Doubled Gloved. If the outer glove shows areas of >black , then the glove integrety has been compromised and I >replace the outer glove. Double gloving can be uncomfortably tight. >I found that a nitrile for the inner glove and a latex for the outer >glove is comfortable for me but it depends on the fit of the gloves >you use . In a pinch I have used two pair of latex. > >c) Any glasswear that I use , I leave in the fume hood overnight for >the fumes to evaporate. > >d) I keep a container of oil in the fume hood when i am working with >the osmium so I can pour oil on a spill if necesary or to neutralise >the osmium if mixed with , say , blood , because it binds to lipids >preferentially. I have used vegetable oil but it goes rancid over time. >Mineral oil is fine. > > > Remember that osmium fumes alone are sometimes used to >fix tissue. So working without proper ventilation means your >eyeballs are going to become fixed. Insist on a proper fume hood. > > > Other than that , it is not bad stuff to work with >:-) > > Margaret > >ps. remember that osmium penetrates only 1 mm in depth so your >tissue has to be diced small. > > >Margaret Horne , >Histology Teaching Assistant, >Dept. of B.SC., >Atlantic Veterinary College, U.P.E.I., >550 University Ave., Charlottetown, >P.E.I., C1A 4P3 >Canada > > >_______________________________________________ >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 d.gregg <@t> juno.com Wed May 26 12:03:19 2004 From: d.gregg <@t> juno.com (d.gregg@juno.com) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Counterstain to use with blue and red substrates Message-ID: <20040526.130320.2888.0.d.gregg@juno.com> Hi all, I am using blue (NBT) and red (AEC) substrates in a double stain and wondered if anyone has a recommendation of a nuclear counterstain that may work with this. I could use metanil yellow a a cytoplasmic stain but would prefer a primarily nuclear stain. I have used Methyl green but it has covered some of the blue in the past. Thanks. Douglas Gregg Plum Island Animal Disease Center Greenport, NY From mhorne <@t> upei.ca Wed May 26 12:39:56 2004 From: mhorne <@t> upei.ca (Margaret Horne) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Osmium Tetroxide Protocol and mineral oil? In-Reply-To: <3.0.6.32.20040526105341.00c0d400@gemini.msu.montana.edu> References: <0556BE8AC5551E4E8AF6BB9E42509BA206EB5C@usca0082k08.labvisi on.apogent.com> Message-ID: <40B4AC3A.28313.15E3E26@localhost> You are right , I replied to Tim but realise that I forgot to send to the rest of the list. I had always used the corn oil , but the last time it was rancid and by boss didn't like the smell. He said the mineral oil would be ok though I was uneasy , but as the rat had been put under we couldn't wait. Tim is right , mineral oil will not work at all. Corn oil is recommended. In one of the web sites he mentioned, there is a mention of using sodium bisulfite for decontamination. Any one ever use that? Margaret Margaret Horne , Histology Teaching Assistant, Dept. of B.SC., Atlantic Veterinary College, U.P.E.I., 550 University Ave., Charlottetown, P.E.I., C1A 4P3 Canada From gcallis <@t> montana.edu Wed May 26 13:24:00 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Counterstain to use with blue and red substrates In-Reply-To: <20040526.130320.2888.0.d.gregg@juno.com> Message-ID: <3.0.6.32.20040526122400.00c167a0@gemini.msu.montana.edu> Personally, we prefer NO counterstain. That way two colors are separate and distinct without the problem you just described. No chromogen is masked by the nuclear staining and any confusion is eliminated. The master of Double staining, Dr Chris van der Loos also does this, and I also suggest you try using VECTOR Blue sometime with AEC, the contrast is superb. At 01:03 PM 5/26/2004 -0400, you wrote: >Hi all, >I am using blue (NBT) and red (AEC) substrates in a double stain and >wondered if anyone has a recommendation of a nuclear counterstain that >may work with this. I could use metanil yellow a a cytoplasmic stain but >would prefer a primarily nuclear stain. I have used Methyl green but it >has covered some of the blue in the past. Thanks. > >Douglas Gregg >Plum Island Animal Disease Center >Greenport, NY > >_______________________________________________ >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 c.m.vanderloos <@t> amc.uva.nl Wed May 26 13:43:38 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] RE: ki67 Message-ID: <1a0b881a1a4e.1a1a4e1a0b88@amc.uva.nl> Dear Veronique, We got very good results using the new rabbit monoclonal Ki67, SP6 from Labvision/Neomarkers. Fix your cryostat sections with buffered formalin for 5 min at room temperature and wash with TBS or PBS. Block endogenous peroxidase and start staining as usual. A two step EnVision or PowerVision detection will do. Acetone-fixation will cause an ugly diffuse staining of the proliferating nuclei. ]Good luck with staining! Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From "Veronique Andriessen" Date Wed, 26 May 2004 14:03:01 +0200 To "Histonet" Subject [Histonet] ki67 Hello histonetters, I'm searching for a Ki67 antibody that will work on frozen sections of rat pancreas. Can you give me some advice? Best regards, Veronique Andriessen BAS Lab. Molecular Liver Cell Biology, Free University Brussels (VUB) Belgium From anissachoi <@t> hotmail.com Wed May 26 14:04:19 2004 From: anissachoi <@t> hotmail.com (Anissa Choi) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Immuno slide problem Message-ID: Thanks to all who replied to my question. Please see if the following will help to gain new insight into my problem. 1. The red box is on the bottom of our slides. I do not think it would interfere with the vortex action of the stainer. If the vortex mixing is not working or weak, I would expect the section on the bottom end of the slide not stained as reagent is dispensed onto the red box area on Ventana stainer. Unlike the Ventana Benchmark with slides lying directly flat on the tray. our slides are clipped on both ends with spaces between slides and the tray. 2.The problem is not repeating. We recut using slides from the same batch and everthing was fine. The other day, out of a run of 20 slides, 2 slides did not work( both control and test did not work).We checked dispenser, AEC kit etc and could not find anything wrong. We repeated without altering anything, same control, same AEC kit, same protease , same antibody dispenser and slides from same batch. The repeated run was fine. 3. Ventana suggested the red paint may get onto some slides and causes patchy and uneven staining. I was told that the particles from the red stuff affect the staining reaction on the Ventana stainer. 4. As I have mentioned in my last e-mail, I put the origional problem slides( the skin section in the middle did not get stained while the control and the other section worked just fine) back on the stainer and ran it again after the stainer was thoroughtly checked and serviced by Ventana. I still could not get the section in the middle stained. 5. Is it reasonable to conclude that we have a slide problem ( occasional defective slides in the batch), rather than a machine problem ? Would you switch to other type of charged slides? Anissa Choi _________________________________________________________________ http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines From GDawson <@t> Milw.Dynacare.com Wed May 26 14:32:53 2004 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] RE: BK virus detection Message-ID: Bill, I use this same antibody for BK virus on IHC. What you need to do is use enzyme rather than heat for epitope retrieval. I use DAKO proteinase K for 4 minutes, 35 minute primary, 30 min. with DAKO Envision+ monoclonal, and 7 minutes of DAKO DAB+. My dilution is at 1:6000. FYI: Don't be shy about demanding a different lot of antibody from this company for their BK virus as I have gotten only 2 out of 4 lots to work. The other 2 lots were a lost cause and I wasted at least 50 slides trying to get them working to no avail. The good antibody lots worked immediately and I just had to fine tune them a bit. Try it out at about 1:2000 first and you should see striking results with BK virus positive kidney. When the antibody is working, it works extremely well. Good Luck, Glen Dawson BS, HT & QIHC (ASCP) Immunohistology Coordinator Milwuakee, WI From ajohnson <@t> aipathology.com Wed May 26 14:44:43 2004 From: ajohnson <@t> aipathology.com (Amy Johnson) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Substitute for Uranyl nitrate Message-ID: <016A64931E1ED511B12C0002B3026080308657@SERV001> Our lab has used up the last of our supply of Uranyl nitrate and need to order it again. I had read an article awhile back that there was a substitute for it, can anyone help us out with this information Thanks Amylin Johnson AIP From ihc <@t> unipathllc.com Wed May 26 15:18:09 2004 From: ihc <@t> unipathllc.com (UniPath IHC) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] RE: BK virus detection In-Reply-To: Message-ID: <000001c4435e$90639a50$4500a8c0@unipath02> Do you know of a source for control slides for JC/BK virus Ab? Brianna Jackson, BS, QIHC UniPath Denver, CO -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dawson, Glen Sent: Wednesday, May 26, 2004 1:33 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: BK virus detection Bill, I use this same antibody for BK virus on IHC. What you need to do is use enzyme rather than heat for epitope retrieval. I use DAKO proteinase K for 4 minutes, 35 minute primary, 30 min. with DAKO Envision+ monoclonal, and 7 minutes of DAKO DAB+. My dilution is at 1:6000. FYI: Don't be shy about demanding a different lot of antibody from this company for their BK virus as I have gotten only 2 out of 4 lots to work. The other 2 lots were a lost cause and I wasted at least 50 slides trying to get them working to no avail. The good antibody lots worked immediately and I just had to fine tune them a bit. Try it out at about 1:2000 first and you should see striking results with BK virus positive kidney. When the antibody is working, it works extremely well. Good Luck, Glen Dawson BS, HT & QIHC (ASCP) Immunohistology Coordinator Milwuakee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kennedya <@t> email.cs.nsw.gov.au Wed May 26 15:47:30 2004 From: kennedya <@t> email.cs.nsw.gov.au (Andrew Kennedy) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] osmium and mineral oil Message-ID: <200405270645697.SM01316@crgcsls813> With regards to using different oils for mopping up Osmium spills, our EM lab use full cream milk powder - it doesn't go rancid and since they only handle small amounts of osmium a small spill can be absorbed by the milk powder and neutralized by the cream content. Simply use in excess and sweep it up with a dustpan and broom for appropriate disposal. Andrew Kennedy Senior Science Officer Anatomical Pathology Concord Repatriation General Hospital Hospital Road Concord NSW 2139 ph: +612 9767 6115 Fax +612 9767 8427 "corpora non agunt nisi fixata" From peptolab <@t> hamptons.com Wed May 26 16:05:19 2004 From: peptolab <@t> hamptons.com (peptolab) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] So sorry, have a cocktail ;-) Message-ID: <002101c44365$27f0b1e0$687bbd18@JEFF> I did not mean to sound accusatory Glen, sorry if I raised your hackles. But I stand by my opinions, which were echoed by some luminaries in this field. As for the law, I don't know but I bet it says that if you are preparing and reading only one set of slides, you better charge for one only, unless perhaps each component can give you different information regarding the patient's tumor using that methodology. Time for a cocktail. Jeff Silverman From convmcm <@t> cc.usu.edu Wed May 26 16:23:51 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Anyone know of a good cartoon showing a basic immuno protocol? In-Reply-To: Message-ID: <000601c44367$be055710$4a737b81@Cygnus> I think Ivan Roitt's website, www.fleshandbones.com/immunology/roitt has a lot of great teaching stuff. You might wan to check it out. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rob Komorowski Sent: Wednesday, May 26, 2004 9:02 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Anyone know of a good cartoon showing a basic immuno protocol? Hi all, I'm trying to find a good step by step cartoon showing a primary antibody linking up with an antigen, a biotinylated secondary antibody, a strepHRP step, and a DAB reaction. I know...sounds specific...but anything close would be helpful for teaching. Thanks! Rob K _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From convmcm <@t> cc.usu.edu Wed May 26 16:27:47 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Substitute for Uranyl nitrate In-Reply-To: <016A64931E1ED511B12C0002B3026080308657@SERV001> Message-ID: <000701c44368$4abe6cf0$4a737b81@Cygnus> Check out the EM suppliers such as EM Science, Ted Pella, PolySciences, etc. TEM still requires the use of UA, so it is still available. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Johnson Sent: Wednesday, May 26, 2004 12:45 PM To: Histonet (E-mail) Subject: [Histonet] Substitute for Uranyl nitrate Our lab has used up the last of our supply of Uranyl nitrate and need to order it again. I had read an article awhile back that there was a substitute for it, can anyone help us out with this information Thanks Amylin Johnson AIP _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From peoshel <@t> wisc.edu Wed May 26 16:39:07 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] test Message-ID: Test. I've had no messages for 2+ days now. Odd. -- Philip Oshel Supervisor, BBPIC microscopy facility 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 JWEEMS <@t> sjha.org Wed May 26 16:38:00 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Steven Slapp Message-ID: Anyone heard from Steven today? Thanks, Joyce 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 convmcm <@t> cc.usu.edu Wed May 26 17:05:27 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Osmium Tetroxide Protocol In-Reply-To: <40B49436.28709.1006818@localhost> Message-ID: <000801c4436d$8d425f50$4a737b81@Cygnus> Margaret Horne Wrote: >>>Double gloving can be uncomfortably tight. I found that a nitrile for the inner glove and a latex for the outer glove is comfortable for me but it depends on the fit of the gloves you use . In a pinch I have used two pair of latex.<<<< Good idea to 2X the gloves, but has anyone checked to see what TYPE of glove is best to use with OsO4? I remember an incident that happened many years ago about someone who was using a heavy metal compound of some kind (mercuric, as I recall). She used latex -- or manybe it was nitrile --gloves. The compound went right through the gloves. It was extremely toxic and the woman died. I wish I could remember all the details of this thing. It made national news. I always think of this when protecting myself from toxic chemicals. Anyway, check the MSDS on OsO4 to see what kind of gloves are best to wear. This also reminds me of the good old days before safety was observed in the labs. When I was learning EM, we handled OsO4, Uranyl nitrate, lead citrate all without gloves. We tried to be very careful, but I can remember getting black on my fingernails and on my fingers from the OsO4. Who knows what else I contaminated myself with... Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Margaret Horne Sent: Wednesday, May 26, 2004 8:57 AM To: histonet@lists.utsouthwestern.edu; mprice26@juno.com Subject: Re: [Histonet] Osmium Tetroxide Protocol Hi , when I handle osmium I always : a) Carry the Osmium to the fume hood in a plastic container . ie. the Osmium solution is in a glass jar with a ground glass stopper and heavily parafilmed. Also the jar is covered in aluminum foil ( to avoid degradation from the light). This jar is Always carried about in a plastic container with a lid. Heavy rubbermaid or tupperware is good. Note that you will see the plastic container turn black over time with the Osmium fumes escaping. We keep it in the " Dirty " fridge , not the " Clean" fridge with our antibodies. b) The plastic container is opened in the fume hood and I wear a labcoat and am Doubled Gloved. If the outer glove shows areas of black , then the glove integrety has been compromised and I replace the outer glove. Double gloving can be uncomfortably tight. I found that a nitrile for the inner glove and a latex for the outer glove is comfortable for me but it depends on the fit of the gloves you use . In a pinch I have used two pair of latex. c) Any glasswear that I use , I leave in the fume hood overnight for the fumes to evaporate. d) I keep a container of oil in the fume hood when i am working with the osmium so I can pour oil on a spill if necesary or to neutralise the osmium if mixed with , say , blood , because it binds to lipids preferentially. I have used vegetable oil but it goes rancid over time. Mineral oil is fine. Remember that osmium fumes alone are sometimes used to fix tissue. So working without proper ventilation means your eyeballs are going to become fixed. Insist on a proper fume hood. Other than that , it is not bad stuff to work with :-) Margaret ps. remember that osmium penetrates only 1 mm in depth so your tissue has to be diced small. Margaret Horne , Histology Teaching Assistant, Dept. of B.SC., Atlantic Veterinary College, U.P.E.I., 550 University Ave., Charlottetown, P.E.I., C1A 4P3 Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From conniegrubaugh <@t> hotmail.com Wed May 26 21:13:42 2004 From: conniegrubaugh <@t> hotmail.com (connie grubaugh) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Steve Slapp Message-ID: Has there been any update on Steve's condition? Connie G. _________________________________________________________________ [1]Is your PC infected? Get a FREE online computer virus scan from McAfee® Security. References 1. http://g.msn.com/8HMAENUS/2755??PS=47575 From mab70 <@t> medschl.cam.ac.uk Thu May 27 02:53:17 2004 From: mab70 <@t> medschl.cam.ac.uk (Margaret Blount) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] exotic species/chief&parietal cells Message-ID: <6C21D947F7189448AB447C57BF2992B4025F16DB@mius.medlan.cam.ac.uk> Dear Sharon, I remembered Harry Cook when you posted your question. He always had very good transparencies when he gave his lectures and I thought he showed us sections of just what you are looking for so I had a look through his book:Histological Demonstration Techniques. Here are the methods he mentions: PAS-Toluidine Blue-Aurantia (Cook, 1962, Stain Technology 37, 317) and Methasol Fast Blue-PAS-Alcian Yellow (Maxwell, 1963, Stain Technology, 38, 286). I haven't tried these, but if Cook recommended them they should be OK. I attach the methods, but I don't know if you can still get all the components. Good luck anyway. Best wishes Margaret Margaret Blount Chief Technician Clinical Biochemistry University of Cambridge Addenbrooke's Hospital Hills Road Cambridge CB2 2QR -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Sent: Wednesday, May 26, 2004 11:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] exotic species/chief&parietal cells Hi all We are looking at the gastric mucosa of the Cape Mole Rat and can not distinguish chief cells on the basis of morphology. There seem to be more parietal cells and we can not be sure of the chief cells. Is there a stain we can use to distinguish between these to cell types as the Hematoxylin & Eosin stain is not making this obvious. Thanks Sharon Marshall Dept. Human Biology University of Cape Town South Africa e-mail: marshall@cormack.uct.ac.za _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.m.vanderloos <@t> amc.uva.nl Thu May 27 05:06:56 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] RE: IHC cartoon 3-step StrepHRP Message-ID: <2631b62653cb.2653cb2631b6@amc.uva.nl> Dear Rob, Separately I send you a cartoon of a 3-step indirect StrepHRP technique. It's a PowerPoint file that also contains a step-by-step animation. Have fun with it! Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Rob Komorowski Date Wed, 26 May 2004 12:02:16 -0400 To Subject [Histonet] Anyone know of a good cartoon showing a basic immuno protocol? Hi all, I'm trying to find a good step by step cartoon showing a primary antibody linking up with an antigen, a biotinylated secondary antibody, a strepHRP step, and a DAB reaction. I know...sounds specific...but anything close would be helpful for teaching. Thanks! Rob K From pengbw <@t> sjtu.edu.cn Thu May 27 05:27:38 2004 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] anti-CD11b Message-ID: <20040527102738.93AD1111B89C@sjtu.edu.cn> Dear all, Would anyone recommend a rat anti mouse CD11b mAb to to identify monocytes, possibly can be used both in IHC and FC. Thanks in advance! Baowei Peng SJTU Shanghai,200030 china From c.m.vanderloos <@t> amc.uva.nl Thu May 27 05:59:47 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] RE: Counterstain to use with blue and red substrates Message-ID: <26b2fe26cda9.26cda926b2fe@amc.uva.nl> Dear Douglas, I presume that the Methyl green that has "covered some of the blue" as you said, was due to the fact that it diffuses away after aqueous mounting needed after using AEC as chromogen. You may try to replace AEC by Vector NovaRed or Romulin Red (BioCare), two peroxidase chromogens that stand organic mounting using VectaMount (a non-xylene based organic mountant) very well. Although the red color is more brownish than with AEC it still contrasts nicely with the dark-blue NBT/BCIP reaction product. Perhaps now your Methyl green counterstain will work for you. Lots of success and let us know! Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From d.gregg@juno.com Date Wed, 26 May 2004 13:03:19 -0400 To histonet@lists.utsouthwestern.edu Subject [Histonet] Counterstain to use with blue and red substrates Hi all, I am using blue (NBT) and red (AEC) substrates in a double stain and wondered if anyone has a recommendation of a nuclear counterstain that may work with this. I could use metanil yellow a a cytoplasmic stain but would prefer a primarily nuclear stain. I have used Methyl green but it has covered some of the blue in the past. Thanks. Douglas Gregg Plum Island Animal Disease Center Greenport, NY From juan.gutierrez <@t> christushealth.org Thu May 27 06:56:51 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Substitute for Uranyl nitrate Message-ID: Phosphomolybdic Acid. The percentage escapes me, either 1%, 5% or 10%. Juan C. Gutierrez, HT(ASCP) Histology Supervisor Christus Santa Rosa Hospital 333 N. Santa Rosa Ave. San Antonio, TX 78207 -----Original Message----- From: Amy Johnson [mailto:ajohnson@aipathology.com] Sent: Wednesday, May 26, 2004 2:45 PM To: Histonet (E-mail) Subject: [Histonet] Substitute for Uranyl nitrate Our lab has used up the last of our supply of Uranyl nitrate and need to order it again. I had read an article awhile back that there was a substitute for it, can anyone help us out with this information Thanks Amylin Johnson AIP _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From qiaolin_deng78 <@t> hotmail.com Thu May 27 07:04:29 2004 From: qiaolin_deng78 <@t> hotmail.com (dengqiaolin deng) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] chicken explant in situ hybridization Message-ID: Dear histonetter: Is there anyone having done whole-mount in situ hybridization for chicken explant? I have tried the same protocol as the one used for whole embryo. But I got some problems caused by the collagen used for explants. I would greatly appreciate someone give me some suggestions. Qiaolin Deng _________________________________________________________________ Tired of spam? Get advanced junk mail protection with MSN 8. http://join.msn.com/?page=features/junkmail From pwg1 <@t> cdc.gov Thu May 27 07:05:01 2004 From: pwg1 <@t> cdc.gov (Greer, Patricia) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Anyone know of a good cartoon showing a basic immunoprotocol? Message-ID: Rob, Look in a catalog that sells antibodies - I know Dakocytomation has graphics in their catalog showing each step in their various kits. Pat Greer Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road Atlanta, GA 30333 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rob Komorowski Sent: Wednesday, May 26, 2004 12:02 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Anyone know of a good cartoon showing a basic immunoprotocol? Hi all, I'm trying to find a good step by step cartoon showing a primary antibody linking up with an antigen, a biotinylated secondary antibody, a strepHRP step, and a DAB reaction. I know...sounds specific...but anything close would be helpful for teaching. Thanks! Rob K _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From m.vankempen <@t> erasmusmc.nl Thu May 27 09:28:08 2004 From: m.vankempen <@t> erasmusmc.nl (m. van mkempen) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Is biotin "inactivated" by fixation with paraformaldehyde? Message-ID: <40B5FAF8.AF9E27F5@erasmusmc.nl> Hello everybody, I have a question about biotin and fixation in paraformaldehyde. The experiment is as follows: Intact lungs from adult mice were labeld with HBSS containing Sulfo-Biotin-X-NHS. The solution was injected through the trachea and the labeling took 15min at 4 degrees Celcius Afterwards the lungs were flushed with PBS/glycin to remove any unreacted biotin The lungs were fixed in 4% paraformaldehyde for 48h Tissue was embedded in parafin and cut into 4 micron sections I used DAKO ABC (streptavidin-HRP) and DAB to detect the biotin labeled proteins. Since Sulfo-Biotin-X-NHS is water soluble I would expect to see a small line of labeled proteins on top of the lung epithelium. But after a few tries I am unable to detect anything. Should I use HIER, switch completely to cryosections or...? One the internet I read that endogenous biotin is partly inactivated by formalin fixation. Could this explain the problems I have? Best regards, Niall Oudesluys (ErasmusMC, Rotterdam, The Netherlands) -- ---------------------------- *- mkempen -* MAILTO:m.vankempen@erasmusmc.nl ----------------------------------------- From Stefan.Wacha <@t> fmi.ch Thu May 27 09:58:34 2004 From: Stefan.Wacha <@t> fmi.ch (Wacha, Stefan) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] microwave fixation Message-ID: Hi Histonetters, I would like to do immunohistochemistry for synaptic markers on sections of dry-ice frozen mouse brains. I heard that the best synaptic staining patterns are achieved with a fixative called NEOFIX combined with 30 secs microwave fixation. Neofix, produced by MERCK, is unfortunately not commercially available anymore and I was not successful to find a replacement. Would anyone have an idea where to get a similar fixative or has a synaptic staining protocol for sections that works better than PFA? Thanks in advance! Stefan Wacha Friedrich Miescher Institute, Basel, Switzerland From michael_lafriniere <@t> memorial.org Thu May 27 10:00:56 2004 From: michael_lafriniere <@t> memorial.org (LaFriniere, Mike) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Steve Slapp Message-ID: Steven has been moved out of intensive care, still in a coma and on respirator. The hospital is Bay State Medical Springfield Mass. Michael LaFriniere -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of connie grubaugh Sent: Wednesday, May 26, 2004 10:14 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Steve Slapp Has there been any update on Steve's condition? Connie G. _________________________________________________________________ [1]Is your PC infected? Get a FREE online computer virus scan from McAfee(r) Security. References 1. http://g.msn.com/8HMAENUS/2755??PS=47575 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Stephen.Eyres <@t> sanofi-synthelabo.com Thu May 27 10:22:29 2004 From: Stephen.Eyres <@t> sanofi-synthelabo.com (Stephen.Eyres@sanofi-synthelabo.com) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Substitute for Uranyl nitrate Message-ID: Hi, Does anyone know of an alternative to cacodylate buffer, which also very toxic. Cheers Steve ---------------------------------------------------------- Le pr?sent message, ainsi que les pi?ces ou annexes qui s'y trouvent ?ventuellement jointes, s'adressent exclusivement ? celles des personnes qu'ils d?signent comme destinataires. Constituant de ce fait une correspondance priv?e ? caract?re confidentiel, leur contenu est prot?g? par le secret des correspondances ?mises, transmises ou re?ues par la voie des t?l?communications. Si ce message ?lectronique vous est parvenu fortuitement, veuillez avoir l'obligeance de le d?truire, puis d'en aviser l'exp?diteur dans les meilleurs d?lais. The Information in this e-mail belongs to Sanofi-Synthelabo, is intended for the use of the individual or entity to which it is addressed, and may contain information that is privileged, confidential, or exempt from disclosure under applicable law. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution, or use of, or reliance on, the contents of this e-mail is prohibited. If you have received this e-mail in error, please notify us immediately by replying back to the sending e-mail address, and delete this e-mail message from your computer. ---------------------------------------------------------- From mab70 <@t> medschl.cam.ac.uk Thu May 27 10:28:43 2004 From: mab70 <@t> medschl.cam.ac.uk (Margaret Blount) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Steve Slapp Message-ID: <6C21D947F7189448AB447C57BF2992B4025F16DC@mius.medlan.cam.ac.uk> Steven, There are lots of prayers winging their way to you. Hold on in there. With love Margaret -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of LaFriniere, Mike Sent: Thursday, May 27, 2004 4:01 PM To: connie grubaugh; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Steve Slapp Steven has been moved out of intensive care, still in a coma and on respirator. The hospital is Bay State Medical Springfield Mass. Michael LaFriniere -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of connie grubaugh Sent: Wednesday, May 26, 2004 10:14 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Steve Slapp Has there been any update on Steve's condition? Connie G. _________________________________________________________________ [1]Is your PC infected? Get a FREE online computer virus scan from McAfee(r) Security. References 1. http://g.msn.com/8HMAENUS/2755??PS=47575 _______________________________________________ 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 mcauliff <@t> umdnj.edu Thu May 27 13:52:51 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Substitute for cacodylate In-Reply-To: References: Message-ID: <40B63903.9020504@umdnj.edu> Phosphate buffers have been around a long time. Lots of different formulas and concentrations. Geoff Stephen.Eyres@sanofi-synthelabo.com wrote: >Hi, > >Does anyone know of an alternative to cacodylate buffer, which also very >toxic. > >Cheers > >Steve > >_______________________________________________ >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 lizchlipala <@t> premierhistology.com Thu May 27 11:02:19 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] anti-CD11b In-Reply-To: <20040527102738.93AD1111B89C@sjtu.edu.cn> Message-ID: <001301c44404$00595930$74d48a80@LIZ> Baowei We have used BD's rat anti mouse CD11b. (Cat Number: 550282). This antibody will only work on frozen sections and unless things have changed over the period of a year I'm not aware of any CD11b marker for mouse that will work on FFPE tissues. If there is anyone out there that knows different please reply. I have a protocol and images if you like. Liz Elizabeth A. Chlipala, BS, HTL(ASCP) 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 MCBD, Room A3B40 Boulder, Colorado 80309 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Baowei Peng Sent: Thursday, May 27, 2004 3:28 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] anti-CD11b Dear all, Would anyone recommend a rat anti mouse CD11b mAb to to identify monocytes, possibly can be used both in IHC and FC. Thanks in advance! Baowei Peng SJTU Shanghai,200030 china From histojock <@t> hotmail.com Thu May 27 11:13:31 2004 From: histojock <@t> hotmail.com (Histo Jock) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Re: Tissue Arrays Message-ID: Making tissue arrays by re-embedding affects staining "down the road" because of basic chemistry. Each time a specimen is exposed to air there will be some loss of some antigens and nucleic acids due to oxidation from atmospheric ozone, acids, etc circulating in the lab. Not all antigens are affected, not every antibody has a problem with it, but it does happen and is well recognized in tissue array labs. Many labs have documented this phenomenon. Most notable is David Rimm's lab at Yale that has made an art out of preserving antigens on tissue arrays by storing them in a special cabinet filled with nitrogen gas. They see dramatic losses in staining intensity in sections left in air for just a few days. The problem with reheating tissue array cores is that this oxidation effect is grossly accelerated by the higher temperatures and air is allowed to penetrate farther into the core once the paraffin has softened from the heat. Some labs have stopped anealing cores into tissue array blocks at 37 degrees because they see a loss in staining. I have had presonal experience with loss of in-situ signal in blocks annealed at 32 degrees versus ones kept at room temp. I remember at least one study that shows slides stored at higher tempuatures (25 degrees, I think) lose some anitgens in fairly short order. If you haven't seen a problem it's probably because of the antibodies you're using. Many polyclonals will do fine, many monoclonals won't. The more the heat+time+exposure to the atmosphere the more the effect. As with everything else in histotechnology it just depends on your specific circumstances. The basic message is that less physical manipulation of a specimen is ALWAYS better than more. There's no reason to heat a specimen to make an array when you can do it perfectly well at room temp. Re-embedded tissue arrays may work great for a lot of things, but experience shows that they do have problems in some applications. HistoJock >Date: Mon, 10 May 2004 04:49:52 +0000 >From: "Thom Jensen" >Subject: Re: [Histonet] Tissue Array >To: Histonet@lists.utsouthwestern.edu >Message-ID: >Content-Type: text/plain > > > How does melting paraffin embedded tissues effect the staining down > the road? That doesn't make since. I have made dozens of multiple > punch arrays by melting the punches and embedding them as you would > normally embed tissues and it has never effected the staining, "DOWN > THE ROAD....." > > Thom >>From: "Histo Jock" >>To: Histonet@lists.utsouthwestern.edu >>Subject: Re: [Histonet] Tissue Array >>Date: Fri, 07 May 2004 20:03:31 -0400 >> >> >>You might want to be careful using Zymed's arrays. I don't think >>they are made with the standard coring method. Rather, I think that >>they use some sort of melting / re-embedding process that can effect >>staining down the road. >> >>HistoJock _________________________________________________________________ Express yourself with the new version of MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From barbara.wright2 <@t> dnax.org Thu May 27 11:41:10 2004 From: barbara.wright2 <@t> dnax.org (Wright, Barbara (SPRI 2)) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] RE: IHC cartoon 3-step StrepHRP Message-ID: <29B25753F6B1D51196110002A589D44401C95B4D@PALMSG30.us.schp.com> Chris, Could you send that same cartoon - would love to see it. Thanks Barb -----Original Message----- From: C.M. van der Loos [mailto:c.m.vanderloos@amc.uva.nl] Sent: Thursday, May 27, 2004 3:07 AM To: histonet@lists.utsouthwestern.edu Cc: komo@bu.edu Subject: [Histonet] RE: IHC cartoon 3-step StrepHRP Dear Rob, Separately I send you a cartoon of a 3-step indirect StrepHRP technique. It's a PowerPoint file that also contains a step-by-step animation. Have fun with it! Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Rob Komorowski Date Wed, 26 May 2004 12:02:16 -0400 To Subject [Histonet] Anyone know of a good cartoon showing a basic immuno protocol? Hi all, I'm trying to find a good step by step cartoon showing a primary antibody linking up with an antigen, a biotinylated secondary antibody, a strepHRP step, and a DAB reaction. I know...sounds specific...but anything close would be helpful for teaching. Thanks! Rob K _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************* 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 convmcm <@t> cc.usu.edu Thu May 27 13:44:55 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Steve Slapp In-Reply-To: Message-ID: <001601c4441a$b49d9080$4a737b81@Cygnus> Thanks, Michael, for that update. I'm so glad to hear that he's moved out of intensive care. I hope that means he'll pull through. Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of LaFriniere, Mike Sent: Thursday, May 27, 2004 8:01 AM To: connie grubaugh; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Steve Slapp Steven has been moved out of intensive care, still in a coma and on respirator. The hospital is Bay State Medical Springfield Mass. Michael LaFriniere -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of connie grubaugh Sent: Wednesday, May 26, 2004 10:14 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Steve Slapp Has there been any update on Steve's condition? Connie G. _________________________________________________________________ [1]Is your PC infected? Get a FREE online computer virus scan from McAfee(r) Security. References 1. http://g.msn.com/8HMAENUS/2755??PS=47575 _______________________________________________ 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 pruegg <@t> colobio.com Thu May 27 14:09:18 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] caspase 3 and pcna In-Reply-To: Message-ID: Jackie and all, could you please advise me on doing caspase 3 and pcna on cells grown on chamber slides, I mostly need advise with caspase, what antibody do you use? Thank you all, Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jackie.O'Connor@abbott.com Sent: Friday, May 21, 2004 8:11 AM To: FreidaC@aol.com Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Embedding I worked in a derm lab for a few years, and it seemed best to embed the skins on a slight angle to the mold, so that when you cut through the soft tissue first, your knife wasn't hitting a straight line of (sometimes) hard epidermis. Also, if you cut through the epidermis first, there is a tendency to damage the knife edge, or drag hard particles through the soft tissue, creating scratch artifacts in your section. Jackie O' FreidaC@aol.com Sent by: histonet-bounces@lists.utsouthwestern.edu 05/21/2004 08:46 AM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Embedding I have a question for all of the derm and hard tissue people out there. How do you embed the sections? Do you cut the dense or hard tissue first or last - or do you embed at an angle? One individual is questioning the answer given in the study guide and so far I have different answers from those I have consulted. Thanks, Freida Carson _______________________________________________ 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 lizchlipala <@t> premierhistology.com Thu May 27 14:20:37 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] caspase 3 and pcna In-Reply-To: Message-ID: <000001c4441f$b4282020$74d48a80@LIZ> Patsy I have done caspase 3 staining on some organotypic cultures (EpiOralT), but these have been processed and embedded into paraffin. The antibody I use is from cell signaling technologies, catalog #9661. I have also used this antibody on rat and mouse specimens. Liz Elizabeth A. Chlipala, BS, HTL(ASCP) 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 MCBD, Room A3B40 Boulder, Colorado 80309 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Thursday, May 27, 2004 12:09 PM To: Jackie.O'Connor@abbott.com Cc: histonet@lists.utsouthwestern.edu Subject: [Histonet] caspase 3 and pcna Jackie and all, could you please advise me on doing caspase 3 and pcna on cells grown on chamber slides, I mostly need advise with caspase, what antibody do you use? Thank you all, Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jackie.O'Connor@abbott.com Sent: Friday, May 21, 2004 8:11 AM To: FreidaC@aol.com Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Embedding I worked in a derm lab for a few years, and it seemed best to embed the skins on a slight angle to the mold, so that when you cut through the soft tissue first, your knife wasn't hitting a straight line of (sometimes) hard epidermis. Also, if you cut through the epidermis first, there is a tendency to damage the knife edge, or drag hard particles through the soft tissue, creating scratch artifacts in your section. Jackie O' FreidaC@aol.com Sent by: histonet-bounces@lists.utsouthwestern.edu 05/21/2004 08:46 AM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Embedding I have a question for all of the derm and hard tissue people out there. How do you embed the sections? Do you cut the dense or hard tissue first or last - or do you embed at an angle? One individual is questioning the answer given in the study guide and so far I have different answers from those I have consulted. Thanks, Freida Carson _______________________________________________ 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 mhorne <@t> upei.ca Thu May 27 12:38:52 2004 From: mhorne <@t> upei.ca (Margaret Horne) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Osmium Tetroxide Protocol In-Reply-To: <000801c4436d$8d425f50$4a737b81@Cygnus> References: <40B49436.28709.1006818@localhost> Message-ID: <40B5FD7B.16379.8435A7@localhost> Hi Connie, Most of the MSDS's don't specify but the second website Tim recommended specifies Nitrile. In fact ,Safeskin Purple Nitrile Xtra Exam Gloves in particular. So I am assuming that the other MSDS's are outdated. Thanks again Tim. Margaret Margaret Horne , Histology Teaching Assistant, Dept. of B.SC., Atlantic Veterinary College, U.P.E.I., 550 University Ave., Charlottetown, P.E.I., C1A 4P3 Canada From pruegg <@t> colobio.com Thu May 27 15:04:37 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] anti-CD11b In-Reply-To: <001301c44404$00595930$74d48a80@LIZ> Message-ID: Liz is right about BD CD11b, I never was able to get it to work on ffpe mouse tissue, I have although had some limited success with DAKO CD68 PGM1 clone which only stains macrophages and monocytes and not all the other myleoid cells, but it is not easy and inconsistant in my hands. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Elizabeth Chlipala Sent: Thursday, May 27, 2004 10:02 AM To: pengbw@sjtu.edu.cn; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] anti-CD11b Baowei We have used BD's rat anti mouse CD11b. (Cat Number: 550282). This antibody will only work on frozen sections and unless things have changed over the period of a year I'm not aware of any CD11b marker for mouse that will work on FFPE tissues. If there is anyone out there that knows different please reply. I have a protocol and images if you like. Liz Elizabeth A. Chlipala, BS, HTL(ASCP) 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 MCBD, Room A3B40 Boulder, Colorado 80309 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Baowei Peng Sent: Thursday, May 27, 2004 3:28 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] anti-CD11b Dear all, Would anyone recommend a rat anti mouse CD11b mAb to to identify monocytes, possibly can be used both in IHC and FC. Thanks in advance! Baowei Peng SJTU Shanghai,200030 china _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From info <@t> instrumedics.com Thu May 27 15:07:00 2004 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Heat and tissue arrays Message-ID: <01af01c44426$2bf79b70$6401a8c0@INSTRUMEDICS22> Histo Jock's explanation of the effect of heat on tissue cores alerts us to the possible loss of antigenicity in cores exposed to heat in the preparation of the recipient block. The reasons certainly seem very plausible. We have discussed the possibility that the water bath can also have an adverse effect on some antigens as well as other cellular molecules.. Bernice Instrumedics From herme013 <@t> umn.edu Thu May 27 17:12:50 2004 From: herme013 <@t> umn.edu (Yves Heremans) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] (no subject) Message-ID: <200405272212.i4RMCo75012964@trojan.software.umn.edu> Hi all, I am looking for a good paraffin embedding protocol for rat liver and subsequent albumin staining. Tissue specimens are about 0.5 cm3. The tissue that I have embedded until now seems too dry: upon sectioning, the tissue disintegrates and the paraffin comes apart from the tissue itself. Soaking in ice water improves the sectioning but I want to avoid having to do this in the future. I use Paramat and the following embedding protocol : fix in 4% PFA for 18-20h at 4°C 1h in 70% ethanol 1h in 80% ethanol 1h in 96% ethanol 2h in 100% ethanol overnight in 100% ethanol 2h in a 1:1 mix of 100% ethanol and histoclear 3h in histoclear overnight in histoclear 2h in a 1:1 mix of histoclear and paramat 3h in paramat 3h in paramat I have to include the overnight incubations since everything is done manually. I have tried shorter dehydration times (30 minutes in 85-95-100 and 100% ethanol, 2 x 30 minutes in histoclear and 3 x 1h in paramat) but without any improvement. 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 From AnthonyH <@t> chw.edu.au Thu May 27 17:15:26 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Steve Slapp Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E1CD@simba.kids> Steve, The Australian Histonetters have you in our thoughts and prayers. We will hear from you soon. 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 ********************************************************************** 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 gcallis <@t> montana.edu Thu May 27 17:40:16 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] anti-CD11b In-Reply-To: <20040527102738.93AD1111B89C@sjtu.edu.cn> Message-ID: <3.0.6.32.20040527164016.00c0d930@gemini.msu.montana.edu> BD Pharmingen has this antibody. You can use it IHC, Immunofluorescent staining and for FACS analysis. Serotec also has rat antiMouse CD11b - depends on who is more convenient for purchase. for At 06:27 PM 5/27/2004 +0800, you wrote: >Dear all, > >Would anyone recommend a rat anti mouse CD11b mAb to to identify monocytes, possibly can be used both in IHC and FC. >Thanks in advance! > >Baowei Peng >SJTU >Shanghai,200030 >china _______________________________________________ >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 degaboh <@t> rice.edu Thu May 27 17:49:34 2004 From: degaboh <@t> rice.edu (ZP) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Osmium Tetroxide Protocol In-Reply-To: <20040527161017.032F719917@fungible5.mail.rice.edu> Message-ID: <20040527224932.679F31DB13@handler2.mail.rice.edu> > Good idea to 2X the gloves, but has anyone checked to see what TYPE of > glove is best to use with OsO4? I remember an incident that happened > many years ago about someone who was using a heavy metal compound of > some kind (mercuric, as I recall). She used latex -- or manybe it was > nitrile --gloves. The compound went right through the gloves. It was > extremely toxic and the woman died. I wish I could remember all the > details of this thing. It made national news. I always think of this > when protecting myself from toxic chemicals. I think I remember that incident also, my organic chem professor was talking about it when the subject of DMSO came up...this lady (I think she was a professor?) was wearing gloves but there was a small hole...and she was working with DMSO and some other compound...the DMSO penetrated her skin with the toxic compound and she died. That's what I remember, but that was more than 6 years ago when I heard about it, so my memory may not be perfect. Zarana Patel degaboh@rice.ed From histo20 <@t> hotmail.com Thu May 27 18:52:36 2004 From: histo20 <@t> hotmail.com (Paula Wilder) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Substitute for Uranyl Nitrate Message-ID: Zinc formalin works well as a substitute Paula Wilder St. Joseph Medical Center Towson, MD 21204 _________________________________________________________________ Learn to simplify your finances and your life in Streamline Your Life from MSN Money. http://special.msn.com/money/0405streamline.armx From tissuearray <@t> hotmail.com Thu May 27 19:07:04 2004 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Heat and Tissue Arrays Message-ID: The only problem we have noticed in a loss of staining is if slides were made months prior to staining. If specimens are left in the paraffin block they seem to be fine when cut just before staining. Often we will make controls ahead and put them in the fridge to help preserve their staining properties. Frozen sections are not an option for us since we as well as all major institutions use paraffin processed blocks for routine specimens. Frozens are only created for special tests and quick diagnosis. It would be expensive if not impossible for us to story thousands of frozens like we are able to store paraffin tissue blocks. Thom Jensen wwww.arrayworkshop.com _________________________________________________________________ [1]Is your PC infected? Get a FREE online computer virus scan from McAfee® Security. References 1. http://g.msn.com/8HMAENUS/2755??PS=47575 From AXavier <@t> mednet.ucla.edu Thu May 27 19:56:36 2004 From: AXavier <@t> mednet.ucla.edu (Xavier, Anton) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Concerning the removal of Massons Trichrome Stain: Message-ID: <85042C7A1A0DD74EACF3145F32F37BA429F9A9@medmail6.mednet.ucla.edu> Dear All, I was wondering if anyone on Histonet knew of a way to remove the Massons Trichrome stain from a slide section? We would like to re-use these particular slides for immunocytochemistry purposes, and thus would like to completely remove the stain without any significant damage to the antigens on the slide. Thanks, Anton Xavier UCLA Vascular Surgery Research. Email: axavier@mednet.ucla.edu From pengbw <@t> sjtu.edu.cn Thu May 27 20:18:50 2004 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] anti-CD11b Message-ID: <20040528011850.BD86310D40AC@sjtu.edu.cn> Thanks you all, In fact, I\'m going to do it on frozen sections. The most prior component I want to stain is monocytes. So maybe I should check DAKO\'s mAb mentioned by Patsy. Baowei Peng From ctsblack <@t> capeheart.uct.ac.za Fri May 28 02:54:34 2004 From: ctsblack <@t> capeheart.uct.ac.za (Melanie Black) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Visualisation of GFP in cells. Message-ID: Hi There Can anybody shed some light on the processing of tissue containing GFP expressing cells for microscopy. Your comments will be appreciated Many Thanks Melanie Black. -- 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 p.vandeplas <@t> aurion.nl Fri May 28 04:02:30 2004 From: p.vandeplas <@t> aurion.nl (Peter van de Plas) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Is biotin "inactivated" by fixation with paraformaldehyde? In-Reply-To: <40B5FAF8.AF9E27F5@erasmusmc.nl> Message-ID: Dear Niall, The effect on antigenicity when fixing biotin with aldehyde depends on the characteristics of the detection system you are using. With the dot-spot test you have a easy approach to test this. Spot a dilution series of antigen (in your case a biotinylated protein) on a strips of nitrocellulose membrane and let dry. Individual strips are now incubated with the fixatives of interest, your control (reference) will be an unfixed strip. Wash in buffer and block free aldehydes in glycine. Remaining protein binding sites on the strip are blocked with BSA. Next step will be the incubation in your Streptavidin-HRP conjugate. Wash an develop the signal. We have a newsletter on this topic available. Please visit our website when you are interested in receiving a copy. In the past I did a comparable test for streptavidin-gold conjugates. I noticed a decrease in labeling density after fixation in paraform. There was only minor effect when using a polyclonal anti-biotin antibody. Hope this info is of help. Peter On Thursday, May 27, 2004, at 04:28 PM, m. van mkempen wrote: > Hello everybody, > > I have a question about biotin and fixation in paraformaldehyde. > > The experiment is as follows: > > Intact lungs from adult mice were labeld with HBSS containing > Sulfo-Biotin-X-NHS. The solution was injected through the trachea and > the labeling took 15min at 4 degrees Celcius > > Afterwards the lungs were flushed with PBS/glycin to remove any > unreacted biotin > > The lungs were fixed in 4% paraformaldehyde for 48h > > Tissue was embedded in parafin and cut into 4 micron sections > > I used DAKO ABC (streptavidin-HRP) and DAB to detect the biotin labeled > proteins. Since Sulfo-Biotin-X-NHS is water soluble I would expect to > see a small line of labeled proteins on top of the lung epithelium. > > But after a few tries I am unable to detect anything. Should I use > HIER, > switch completely to cryosections or...? > > One the internet I read that endogenous biotin is partly inactivated by > formalin fixation. Could this explain the problems I have? > > Best regards, Niall Oudesluys (ErasmusMC, Rotterdam, The Netherlands) > > ----------- Peter van de Plas Aurion Costerweg 5 6702 AA Wageningen The Netherlands Phone: +31 317 497676 Fax: +31 317 415955 http://www.aurion.nl/ From ian.montgomery <@t> bio.gla.ac.uk Fri May 28 04:02:54 2004 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:23:02 2005 Subject: Fwd: RE: [Histonet] Substitute for Uranyl nitrate Message-ID: <6.1.0.6.2.20040528100042.02d35190@udcf.gla.ac.uk> Steve, Any phosphate buffer. Remember the final ultrastructure depends on the buffer of choice so make sure you choose correctly. Ian. >Hi, > >Does anyone know of an alternative to cacodylate buffer, which also very >toxic. > >Cheers > >Steve > >---------------------------------------------------------- >Le pr?sent message, ainsi que les pi?ces ou annexes qui s'y trouvent >?ventuellement jointes, s'adressent exclusivement ? celles des personnes >qu'ils d?signent comme destinataires. Constituant de ce fait une >correspondance priv?e ? caract?re confidentiel, leur contenu est prot?g? >par le secret des correspondances ?mises, transmises ou re?ues par la voie >des t?l?communications. Si ce message ?lectronique vous est parvenu >fortuitement, veuillez avoir l'obligeance de le d?truire, puis d'en aviser >l'exp?diteur dans les meilleurs d?lais. > >The Information in this e-mail belongs to Sanofi-Synthelabo, is intended >for the use of the individual or entity to which it is addressed, and may >contain information that is privileged, confidential, or exempt from >disclosure under applicable law. If you are not the intended recipient, you >are hereby notified that any disclosure, copying, distribution, or use of, >or reliance on, the contents of this e-mail is prohibited. If you have >received this e-mail in error, please notify us immediately by replying >back to the sending e-mail address, and delete this e-mail message from >your computer. >---------------------------------------------------------- > > >_______________________________________________ >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 t-sherman <@t> comcast.net Fri May 28 05:14:35 2004 From: t-sherman <@t> comcast.net (Todd Sherman) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Is biotin "inactivated" by fixation with paraformaldehyde? Message-ID: <40B7110B.6000601@comcast.net> -----BEGIN PGP SIGNED MESSAGE----- Hash: SHA1 Dear Niall, I'm not familiar with the detection system you have described, i.e. membrane-impermeant HBSS/Sulfo-Biotin-X-NHS (Excuse my ignorance but what is HBSS?) but the incubation time and temperature seems extremely brief? Is that what the product insert recommends or what some other lab has used successfully? Have you had success with this previously? Is +4deg C the optimum temperature of reactivity? The lung temperature will be elevated for a while until the reagent gets flushed through the entire system and equilibrates. Do you begin timing as soon as reagents enter the trachea or when the reagent has had a chance to reach the most terminal bronchioles? Did you detect any "staining" at the most proximal levels, i.e. sections where the reagent had the most time to bind, or were bronchus to terminal bronchioles equally devoid of DAB complex? Or another question: Were pulmonary secretions (mucous) flushed out prior to incubation with HBSS/Sulfo-Biotin-X-NHS? Maybe the secretions served as a physical barrier to your incubate. I know I'm not specifically answering your aldehyde issue but these other issues pop into my head first. You'll just have to excuse my ignorance. Now as long as there is free amine, then the aldehyde can bind and, I guess, deter subsequent binding by streptavidin-HRP. So if the Sulfo-Biotin-X-NHS has free unbound amine, then methylene-bridge formation will continue. I just don't know the binding sequence of the reagents involved. Is this the theoretical model? epithelia->HBSS->(Sulfo-Biotin-X-NHS)->xaldehyde ~ | ~ v ~ Streptavidin ~ | ~ HRP ~ | ~ v ~ DAB Which is the "lumenal end" of the Sulfo-Biotin-X-NHS complex? What does HBSS bind to? Hope this helps, Todd Todd Sherman President HistoSoft Corporation "Biology in a new form..." Home: www.histosoft.com Member Services: www.myhistosoft.com histonet-request@lists.utsouthwestern.edu wrote: | Today's Topics: | 23. Is biotin "inactivated" by fixation with paraformaldehyde? | (m. van mkempen) | | ---------------------------------------------------------------------- | Message: 23 Date: Thu, 27 May 2004 16:28:08 +0200 From: "m. van mkempen" Subject: [Histonet] Is biotin "inactivated" by fixation with paraformaldehyde? To: Histonetlist Message-ID: <40B5FAF8.AF9E27F5@erasmusmc.nl> Content-Type: text/plain; charset=us-ascii Hello everybody, I have a question about biotin and fixation in paraformaldehyde. The experiment is as follows: Intact lungs from adult mice were labeld with HBSS containing Sulfo-Biotin-X-NHS. The solution was injected through the trachea and the labeling took 15min at 4 degrees Celcius Afterwards the lungs were flushed with PBS/glycin to remove any unreacted biotin The lungs were fixed in 4% paraformaldehyde for 48h Tissue was embedded in parafin and cut into 4 micron sections I used DAKO ABC (streptavidin-HRP) and DAB to detect the biotin labeled proteins. Since Sulfo-Biotin-X-NHS is water soluble I would expect to see a small line of labeled proteins on top of the lung epithelium. But after a few tries I am unable to detect anything. Should I use HIER, switch completely to cryosections or...? One the internet I read that endogenous biotin is partly inactivated by formalin fixation. Could this explain the problems I have? Best regards, Niall Oudesluys (ErasmusMC, Rotterdam, The Netherlands) - -- ---------------------------- *- mkempen -* MAILTO:m.vankempen@erasmusmc.nl -----BEGIN PGP SIGNATURE----- Version: GnuPG v1.2.3 (MingW32) Comment: Using GnuPG with MultiZilla - http://enigmail.mozdev.org iD8DBQFAtxEKEmHXdrslGRcRAhcnAJ0dOv0YhoVOkCWnRQojeU14IVpuxACdFU+I bmgJ+tukvbu8u3AW8itsUIs= =VkSg -----END PGP SIGNATURE----- From David.Edmondson <@t> christie-tr.nwest.nhs.uk Fri May 28 05:17:43 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Concerning the removal of Massons Trichrome Stain: Message-ID: Anton If you are using heat mediated antigen retrieval then that will probably do the business and take out all the staining without doing anything untoward Dave Christie's Manchester UK -----Original Message----- From: Xavier, Anton [mailto:AXavier@mednet.ucla.edu] Sent: 28 May 2004 01:57 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Concerning the removal of Massons Trichrome Stain: Dear All, I was wondering if anyone on Histonet knew of a way to remove the Massons Trichrome stain from a slide section? We would like to re-use these particular slides for immunocytochemistry purposes, and thus would like to completely remove the stain without any significant damage to the antigens on the slide. Thanks, Anton Xavier UCLA Vascular Surgery Research. Email: axavier@mednet.ucla.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Fri May 28 07:31:47 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Santa Cruz Murine Osteocalcin Message-ID: Happy Friday before a holiday weekend everyone - well, in the US anyway. Happy regular Friday to everyone else! I'm in the throes of a serious migraine, but have to work up an osteocalcin antibody today. Can anyone take pity on me and provide me with a protocol for murine osteoblasts in decalcified paraffin embedded tibias? If you only knew how bad my head hurts . . . . . Jackie O' From DKUMISKI <@t> mail.mcg.edu Fri May 28 07:40:49 2004 From: DKUMISKI <@t> mail.mcg.edu (Donna Kumiski) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Counterstain to use with blue and red substrates Message-ID: What about using a fluorescent counter stain such as DAPI? You would of course have to have the correct filter to view it. I have included a link with the details of the stain. Donna http://www.arches.uga.edu/~apollo/InnoGenex%20Datasheet.htm ---------------------------------------------- Donna Kumiski HT (ASCP) Medical College of Georgia Dept. of Cellular Biology and Anatomy CB 1202 Augusta, Georgia 30912-2000 (706) 721-6278 dkumiski@mail.mcg.edu ----------------------------------------------- From t-sherman <@t> comcast.net Fri May 28 08:04:00 2004 From: t-sherman <@t> comcast.net (Todd Sherman) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Immuno slide problem Message-ID: <40B738C0.8080704@comcast.net> -----BEGIN PGP SIGNED MESSAGE----- Hash: SHA1 Anissa, Short answer, slides sound suspect. More elaboration below ****. From: "Anissa Choi" Thanks to all who replied to my question. Please see if the following will help to gain new insight into my problem. **** **** 2.The problem is not repeating. We recut using slides from the same batch and everything was fine. The other day, out of a run of 20 slides, 2 slides did not work( both control and test did not work). We checked dispenser, AEC kit etc and could not find anything wrong. We repeated without altering anything, same control, same AEC kit, same protease, same antibody dispenser and slides from same batch. The repeated run was fine. ****Assuming you had the same number of slides loaded, i.e. a bad slot (or slots) on the machine was not excluded from the follow-up run, it sounds like a bad batch of slides. **** 3. Ventana suggested the red paint may get onto some slides and causes patchy and uneven staining. I was told that the particles from the red stuff affect the staining reaction on the Ventana stainer. ****Its inconsistency would refute this argument if it was exclusively the red "paint" that was hindering the staining. Conceivably, Ventana's hypothesis is possible but the intermittent irregularity seems weird and inconclusive. May be a sealant or "post-painting" process in the manufacture of these slides that is leaving trace chemicals - so it's not the paint but something else coating the slides that is creating "non-stain zones", for lack of a better description. **** **** **** 5. Is it reasonable to conclude that we have a slide problem (occasional defective slides in the batch), rather than a machine problem ? Would you switch to other type of charged slides? ****I would try several brands of charged slides and run them concurrently with only positive and negative controls. Forget real specimens until you get this resolved. Record the slot number of positives that do not stain (assuming some of them do not stain). You may even need to try a very common, universal Ab that detects a very common antigen present in all tissues to ensure that your Ab doesn't introduce another set of issues. If another batch of slides results in this irregular pattern, then it almost has to be the Ventana. **** Hope this helps, Todd Todd Sherman President HistoSoft Corporation "Biology in a new form..." Home: www.histosoft.com Member Services: www.myhistosoft.com -----BEGIN PGP SIGNATURE----- Version: GnuPG v1.2.3 (MingW32) Comment: Using GnuPG with MultiZilla - http://enigmail.mozdev.org iD8DBQFAtzi/EmHXdrslGRcRAuuFAJ9JMBp2HdNGcL2qIFHsxMTCJh61lQCgs+0d o34yUZxPK9pL4vO+dHwMfYg= =uqCA -----END PGP SIGNATURE----- From hmcleod <@t> chempath.uct.ac.za Fri May 28 08:47:40 2004 From: hmcleod <@t> chempath.uct.ac.za (Mcleod) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] HYALURONIDASE Message-ID: <40B742FC.E6D86013@chempath.uct.ac.za> Dear All We need to order hyaluronidase. We currently are using a MERCK PRODUCT (K 27072808) which (acording to our local agency) is no longer available from Merck. Whose product do histonetters recommend .... ...and which of the Sigma products would you recommend? Thankyou for all responses Heather From mprice26 <@t> juno.com Fri May 28 09:11:45 2004 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] RE: Procedure for Sulfated Alcian Blue Message-ID: <20040528.071240.23103.115300@webmail18.nyc.untd.com> Histonetters, Does anyone have a procedure for "Sulfated Alcian Blue" they could share with me? Thank you. Marsha ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! From lchung <@t> ppmh.org Fri May 28 09:38:08 2004 From: lchung <@t> ppmh.org (Chung, Luong) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] snap freeze with liquid nitrogen procedure Message-ID: all, Anyone has a procedure for liquid nitrogen snap freeze tissue? Do you use talc and where can I get some of that? Thanks in advance for any advice. Bruce From tflore <@t> lsuhsc.edu Fri May 28 09:44:01 2004 From: tflore <@t> lsuhsc.edu (Flores, Teresa) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] snap freeze with liquid nitrogen procedure Message-ID: What technique is being performed on what type of tissues? Teresa -----Original Message----- From: Chung, Luong [mailto:lchung@ppmh.org] Sent: Friday, May 28, 2004 9:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] snap freeze with liquid nitrogen procedure all, Anyone has a procedure for liquid nitrogen snap freeze tissue? Do you use talc and where can I get some of that? Thanks in advance for any advice. Bruce _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JMahoney <@t> alegent.org Fri May 28 09:45:05 2004 From: JMahoney <@t> alegent.org (Janice A Mahoney) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Re: Tissue Arrays Message-ID: I'd be interested in knowing if anyone has had an issue with false positive staining of Her-2-neu on breast needle biopsies. Any ideas on why this happens? Thanks, Jan Janice Mahoney Histology/Cytology Coordinator Alegent Health Laboratory Phone (402)717-2889 Fax (402)717-5231 "There's NO excuse for Domestic Violence" Crisis Lines: Omaha YWCA 402-345-7273 Sarpy County 1-800-523-3666 Council Bluffs 1-712-328-0266 NE's Statewide 1-800-876-6238 >>> "Histo Jock" 05/27/2004 11:13:31 AM >>> Making tissue arrays by re-embedding affects staining "down the road" because of basic chemistry. Each time a specimen is exposed to air there will be some loss of some antigens and nucleic acids due to oxidation from atmospheric ozone, acids, etc circulating in the lab. Not all antigens are affected, not every antibody has a problem with it, but it does happen and is well recognized in tissue array labs. Many labs have documented this phenomenon. Most notable is David Rimm's lab at Yale that has made an art out of preserving antigens on tissue arrays by storing them in a special cabinet filled with nitrogen gas. They see dramatic losses in staining intensity in sections left in air for just a few days. The problem with reheating tissue array cores is that this oxidation effect is grossly accelerated by the higher temperatures and air is allowed to penetrate farther into the core once the paraffin has softened from the heat. Some labs have stopped anealing cores into tissue array blocks at 37 degrees because they see a loss in staining. I have had presonal experience with loss of in-situ signal in blocks annealed at 32 degrees versus ones kept at room temp. I remember at least one study that shows slides stored at higher tempuatures (25 degrees, I think) lose some anitgens in fairly short order. If you haven't seen a problem it's probably because of the antibodies you're using. Many polyclonals will do fine, many monoclonals won't. The more the heat+time+exposure to the atmosphere the more the effect. As with everything else in histotechnology it just depends on your specific circumstances. The basic message is that less physical manipulation of a specimen is ALWAYS better than more. There's no reason to heat a specimen to make an array when you can do it perfectly well at room temp. Re-embedded tissue arrays may work great for a lot of things, but experience shows that they do have problems in some applications. HistoJock >Date: Mon, 10 May 2004 04:49:52 +0000 >From: "Thom Jensen" >Subject: Re: [Histonet] Tissue Array >To: Histonet@lists.utsouthwestern.edu >Message-ID: >Content-Type: text/plain > > > How does melting paraffin embedded tissues effect the staining down > the road? That doesn't make since. I have made dozens of multiple > punch arrays by melting the punches and embedding them as you would > normally embed tissues and it has never effected the staining, "DOWN > THE ROAD....." > > Thom >>From: "Histo Jock" >>To: Histonet@lists.utsouthwestern.edu >>Subject: Re: [Histonet] Tissue Array >>Date: Fri, 07 May 2004 20:03:31 -0400 >> >> >>You might want to be careful using Zymed's arrays. I don't think >>they are made with the standard coring method. Rather, I think that >>they use some sort of melting / re-embedding process that can effect >>staining down the road. >> >>HistoJock _________________________________________________________________ Express yourself with the new version of MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri May 28 10:51:52 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] anti-CD11b In-Reply-To: <20040528011850.BD86310D40AC@sjtu.edu.cn> Message-ID: <3.0.6.32.20040528095152.00c12830@gemini.msu.montana.edu> Patsy wrote: I have although had some limited success with DAKO CD68 PGM1 clone which only stains macrophages and monocytes and not all the other myleoid cells, but it is not easy and inconsistant in my hands. Caveat: This is a mouse antiHuman CD68 for macrophages, and you will run into the problem of staining with mouse antibody on mouse tissue, a whole other problem introduced requiring mouse on mouse kits and adds to expense of procedure. You can avoid this problem by using CD11b aka Mac1 (Clone M1/70) Rat antiMouse Monoclonal from BD Pharmingen. Do a dilution panel starting at 10 ug/ml, normally you can have good staining at 1:500 - 1:1000 or more depending on the chromogen used, primary has original conc of 0.5mg/ml. and incubated for 30 min at room temperature. Be sure to use the isotype matched control or rat IgG as a negative control We have superb success with acetone(75ml/absolute ethanol (25 ml) fixation. Make sure you air dry the frozen sections OVERNIGHT AT ROOM TEMPERATURE, then fix in this fixative for 5 min at Room temperature, do not dry after fixation but go immediately to 3 changes of PBS (Dulbeccos) and proceed to other blocking steps (DAKO peroxidase block for frozen sections), biotin blocking (Vector), followed by IHC. For a secondary antibody, Goat antiRat bioitinylated, adsorbed to mouse and is an F(ab')2 frag, from Biosource or a Donkey antiRat biotinylated, F(ab')2 frag of IgG adsorbed to Mouse biotinylated from Jackson Immunoresearch is recommended. This secondary should be diluted in the normal serum block containing mouse serum (mouse serum concentration varies from 1 - 5%, we take the middle concentration of 2.5%). We also prefer to use this antibody biotinylated - this eliminates the secondary antibody, and you go directly to Strepavidin-HRP for 20 min. We do not use kits, and our Strepavidin-HRP comes from Biosource (0.5 to 0.6mg/ml diluted 1:500). For a block, we prefer normal serum 10% goat or Donkey (matched to host of secondary) with 2.5% mouse serum (heat inactivated) - Our morphology is excellent, better than acetone fixed frozen sections, and our results are very clean, without any background staining. Make sure you positive control is from an animal stimulated to produce macrophages in order to control the chromogen staining with a microscope. At 09:18 AM 5/28/2004 +0800, you wrote: >Thanks you all, >In fact, I\'m going to do it on frozen sections. >The most prior component I want to stain is monocytes. >So maybe I should check DAKO\'s mAb mentioned by Patsy. > >Baowei Peng _______________________________________________ >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 Fri May 28 11:04:07 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Counterstain to use with blue and redsubstrates In-Reply-To: Message-ID: <3.0.6.32.20040528100407.00c12830@gemini.msu.montana.edu> One can purchase Vectashield containing DAPI, it is aqueous mounting media (also comes in a form that sets up hard!), coverslip and stain with DAPI all at once! Very tidy. If you use VECTOR red as a chromogen, it fluoresces with UV light AND appears red via light microscopy. To have crisp Vector Red (I think the new DAKO red chromogen also fluoresces) be sure you follow brochure directions AND add Tween 20 to buffer when making up Vector Red. The best of both worlds. At 08:40 AM 5/28/2004 -0400, you wrote: >What about using a fluorescent counter stain such as DAPI? You would of >course have to > have the correct filter to view it. I have included a link with the >details of the stain. > Donna > > > http://www.arches.uga.edu/~apollo/InnoGenex%20Datasheet.htm > >---------------------------------------------- >Donna Kumiski HT (ASCP) >Medical College of Georgia >Dept. of Cellular Biology and Anatomy >CB 1202 >Augusta, Georgia 30912-2000 >(706) 721-6278 > dkumiski@mail.mcg.edu >----------------------------------------------- > > >_______________________________________________ >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 mab70 <@t> medschl.cam.ac.uk Fri May 28 11:23:07 2004 From: mab70 <@t> medschl.cam.ac.uk (Margaret Blount) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Counterstain to use with blue and redsubstrates Message-ID: <6C21D947F7189448AB447C57BF2992B4025F16E2@mius.medlan.cam.ac.uk> Sigma Fast Fast Red TR/maphthol AS-MX phosphate also fluoresces red. In case you are a Sigma fan. Margaret Margaret Blount Chief Technician Clinical Biochemistry University of Cambridge Addenbrooke's Hospital Hills Road Cambridge CB2 2QR -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gayle Callis Sent: Friday, May 28, 2004 5:04 PM To: Donna Kumiski; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Counterstain to use with blue and redsubstrates One can purchase Vectashield containing DAPI, it is aqueous mounting media (also comes in a form that sets up hard!), coverslip and stain with DAPI all at once! Very tidy. If you use VECTOR red as a chromogen, it fluoresces with UV light AND appears red via light microscopy. To have crisp Vector Red (I think the new DAKO red chromogen also fluoresces) be sure you follow brochure directions AND add Tween 20 to buffer when making up Vector Red. The best of both worlds. At 08:40 AM 5/28/2004 -0400, you wrote: >What about using a fluorescent counter stain such as DAPI? You would of >course have to > have the correct filter to view it. I have included a link with the >details of the stain. > Donna > > > http://www.arches.uga.edu/~apollo/InnoGenex%20Datasheet.htm > >---------------------------------------------- >Donna Kumiski HT (ASCP) >Medical College of Georgia >Dept. of Cellular Biology and Anatomy >CB 1202 >Augusta, Georgia 30912-2000 >(706) 721-6278 > dkumiski@mail.mcg.edu >----------------------------------------------- > > >_______________________________________________ >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 CKByrne <@t> labvision.com Fri May 28 11:36:36 2004 From: CKByrne <@t> labvision.com (Kyle-Byrne, Carrie - Labvision) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Immuno slide problem Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20A9712@usca0082k08.labvision.apogent.com> I think most everyone (or I hope they are) is aware the Erie Scientific makes the majority (but certainly not all) of the available charged slides on the market in the US. The slides sold by Ventana are manufactured by Erie. I would suggest calling them and asking if they've had any other reports of problems with they type of slide you are using. Carrie Kyle-Byrne Sr. Research Assoc., Pathology Lab Lab Vision, Corp. 47790 Westinghouse Ave. Fremont, CA 94539 ckbyrne@labvision.com www.labvision.com -----Original Message----- From: Todd Sherman [mailto:t-sherman@comcast.net] Sent: Friday, May 28, 2004 6:04 AM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Immuno slide problem -----BEGIN PGP SIGNED MESSAGE----- Hash: SHA1 Anissa, Short answer, slides sound suspect. More elaboration below ****. From: "Anissa Choi" Thanks to all who replied to my question. Please see if the following will help to gain new insight into my problem. **** **** 2.The problem is not repeating. We recut using slides from the same batch and everything was fine. The other day, out of a run of 20 slides, 2 slides did not work( both control and test did not work). We checked dispenser, AEC kit etc and could not find anything wrong. We repeated without altering anything, same control, same AEC kit, same protease, same antibody dispenser and slides from same batch. The repeated run was fine. ****Assuming you had the same number of slides loaded, i.e. a bad slot (or slots) on the machine was not excluded from the follow-up run, it sounds like a bad batch of slides. **** 3. Ventana suggested the red paint may get onto some slides and causes patchy and uneven staining. I was told that the particles from the red stuff affect the staining reaction on the Ventana stainer. ****Its inconsistency would refute this argument if it was exclusively the red "paint" that was hindering the staining. Conceivably, Ventana's hypothesis is possible but the intermittent irregularity seems weird and inconclusive. May be a sealant or "post-painting" process in the manufacture of these slides that is leaving trace chemicals - so it's not the paint but something else coating the slides that is creating "non-stain zones", for lack of a better description. **** **** **** 5. Is it reasonable to conclude that we have a slide problem (occasional defective slides in the batch), rather than a machine problem ? Would you switch to other type of charged slides? ****I would try several brands of charged slides and run them concurrently with only positive and negative controls. Forget real specimens until you get this resolved. Record the slot number of positives that do not stain (assuming some of them do not stain). You may even need to try a very common, universal Ab that detects a very common antigen present in all tissues to ensure that your Ab doesn't introduce another set of issues. If another batch of slides results in this irregular pattern, then it almost has to be the Ventana. **** Hope this helps, Todd Todd Sherman President HistoSoft Corporation "Biology in a new form..." Home: www.histosoft.com Member Services: www.myhistosoft.com -----BEGIN PGP SIGNATURE----- Version: GnuPG v1.2.3 (MingW32) Comment: Using GnuPG with MultiZilla - http://enigmail.mozdev.org iD8DBQFAtzi/EmHXdrslGRcRAuuFAJ9JMBp2HdNGcL2qIFHsxMTCJh61lQCgs+0d o34yUZxPK9pL4vO+dHwMfYg= =uqCA -----END PGP SIGNATURE----- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Fri May 28 11:51:42 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Immuno slide problem Message-ID: Hmmmm - does anyone know who makes charged slides for Lab Storage Systems? I just bought a ton of them because I was sucked in by the pretty pretty colors.....still have that migraine.... Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotherapeutics "Kyle-Byrne, Carrie - Labvision" Sent by: histonet-bounces@lists.utsouthwestern.edu 05/28/2004 11:36 AM To: "'histonet@lists.utsouthwestern.edu'" cc: Subject: RE: [Histonet] Immuno slide problem I think most everyone (or I hope they are) is aware the Erie Scientific makes the majority (but certainly not all) of the available charged slides on the market in the US. The slides sold by Ventana are manufactured by Erie. I would suggest calling them and asking if they've had any other reports of problems with they type of slide you are using. Carrie Kyle-Byrne Sr. Research Assoc., Pathology Lab Lab Vision, Corp. 47790 Westinghouse Ave. Fremont, CA 94539 ckbyrne@labvision.com www.labvision.com -----Original Message----- From: Todd Sherman [mailto:t-sherman@comcast.net] Sent: Friday, May 28, 2004 6:04 AM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Immuno slide problem -----BEGIN PGP SIGNED MESSAGE----- Hash: SHA1 Anissa, Short answer, slides sound suspect. More elaboration below ****. From: "Anissa Choi" Thanks to all who replied to my question. Please see if the following will help to gain new insight into my problem. **** **** 2.The problem is not repeating. We recut using slides from the same batch and everything was fine. The other day, out of a run of 20 slides, 2 slides did not work( both control and test did not work). We checked dispenser, AEC kit etc and could not find anything wrong. We repeated without altering anything, same control, same AEC kit, same protease, same antibody dispenser and slides from same batch. The repeated run was fine. ****Assuming you had the same number of slides loaded, i.e. a bad slot (or slots) on the machine was not excluded from the follow-up run, it sounds like a bad batch of slides. **** 3. Ventana suggested the red paint may get onto some slides and causes patchy and uneven staining. I was told that the particles from the red stuff affect the staining reaction on the Ventana stainer. ****Its inconsistency would refute this argument if it was exclusively the red "paint" that was hindering the staining. Conceivably, Ventana's hypothesis is possible but the intermittent irregularity seems weird and inconclusive. May be a sealant or "post-painting" process in the manufacture of these slides that is leaving trace chemicals - so it's not the paint but something else coating the slides that is creating "non-stain zones", for lack of a better description. **** **** **** 5. Is it reasonable to conclude that we have a slide problem (occasional defective slides in the batch), rather than a machine problem ? Would you switch to other type of charged slides? ****I would try several brands of charged slides and run them concurrently with only positive and negative controls. Forget real specimens until you get this resolved. Record the slot number of positives that do not stain (assuming some of them do not stain). You may even need to try a very common, universal Ab that detects a very common antigen present in all tissues to ensure that your Ab doesn't introduce another set of issues. If another batch of slides results in this irregular pattern, then it almost has to be the Ventana. **** Hope this helps, Todd Todd Sherman President HistoSoft Corporation "Biology in a new form..." Home: www.histosoft.com Member Services: www.myhistosoft.com -----BEGIN PGP SIGNATURE----- Version: GnuPG v1.2.3 (MingW32) Comment: Using GnuPG with MultiZilla - http://enigmail.mozdev.org iD8DBQFAtzi/EmHXdrslGRcRAuuFAJ9JMBp2HdNGcL2qIFHsxMTCJh61lQCgs+0d o34yUZxPK9pL4vO+dHwMfYg= =uqCA -----END PGP SIGNATURE----- _______________________________________________ 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 brett_connolly <@t> merck.com Fri May 28 12:22:04 2004 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Re: slide problem Message-ID: I have never had a problem with Starfrost adhesive slides from Mercedes Medical. My understanding is they are manufactured in Germany...and not by Erie. Great slides! http://www.mercedesmedical.com/UnloadBoxcar3of3.cfm?DeptID=2&CatID=51&SubCat ID=185&ProdGroupID=310 Call Chad Brown 800-331-2716 x209 Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP26A-3000 PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-652-2075 e-mail. brett_connolly@merck.com ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ From brett_connolly <@t> merck.com Fri May 28 12:40:48 2004 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Re: slide problem Message-ID: Guess the link didn't go through correctly... www.mercedesmedical.com Go to lab supplies, then microscope slides Brett -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Connolly, Brett M Sent: Friday, May 28, 2004 1:22 PM To: 'HISTONET' (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Re: slide problem I have never had a problem with Starfrost adhesive slides from Mercedes Medical. My understanding is they are manufactured in Germany...and not by Erie. Great slides! http://www.mercedesmedical.com/UnloadBoxcar3of3.cfm?DeptID=2&CatID=51&SubCat ID=185&ProdGroupID=310 Call Chad Brown 800-331-2716 x209 Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP26A-3000 PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-652-2075 e-mail. brett_connolly@merck.com ---------------------------------------------------------------------------- -- Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ---------------------------------------------------------------------------- -- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------------------------- -- Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ---------------------------------------------------------------------------- -- ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ From Traczyk7 <@t> aol.com Fri May 28 13:36:58 2004 From: Traczyk7 <@t> aol.com (Traczyk7@aol.com) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Steve Slap Message-ID: <102.466a0a69.2de8e0ca@aol.com> So many of you have asked about Steve Slap that I thought the best way to get the word out is through the net. As many of you have already heard, Steve was in accident while riding his Honda scooter. No one saw what actually happened, but thank God he was found quickly. It happened near his home on May 11th and he was airlifted to the trauma center in Springfield. I spoke with Lisa (Steve's significant other) just this morning. Steve is still in a coma and in ICU. For now, it's just a waiting game. The doctor's are optimistic that he will come out of the coma but have given Lisa no time frame. They say he is otherwise healthy and that will work well for the long recovery process. When or if he actually gets back on his feet is not known because there is brain stem damage. If there is any change I will let you all know. The best we can do now is to keep Steve and his family in our prayers. Lisa did say she would love to hear from Steve's friends. So if you would like to send a card, that would be great. Include a picture, if you have one, with who and what it's about written on the back. Part of the healing process is to just keep talking to Steve. Cards and pictures will give Lisa and the nurses different things to talk about with him. Steve's son Jeremy is holding his own. He's a good kid. Their address is: Lisa Smith & Steve Slap PO Box 31 Lake Pleasant, MA 01347 I will pass along any greetings posted to HistoNet. If you prefer, you can email me directly at traczyk7@aol.com Regards, Dorothy Murphy Traczyk Hacker Instruments & Industries Inc. From cgenty <@t> bcm.tmc.edu Fri May 28 14:23:43 2004 From: cgenty <@t> bcm.tmc.edu (Genty, Carlos E) Date: Fri Sep 16 15:23:02 2005 Subject: [Histonet] Tissue Arrays (TMA) Message-ID: <797AB4E975D23946B86A2AA63965195720B5C1@BCMEVS1.ad.bcm.edu> Histo Jock, Interesting post. Oxidation is well documented and I agree with you on the fact that it does not affect all antigens equally. We also store our samples cold in order to slow/arrest the process. Do you have any data or perhaps a reference to support your claim that this happens when annealing cores to a recipient array block? How do you see a reduction, is it on the outer edge of the core or is it affected evenly over the entire surface? We test hundreds of antibodies, with and without TMAs and while oxidation on pre-cut slides had been a problem in the past, we have not experienced this with the TMA blocks. Best regards, Carlos Genty Baylor College of Medicine -----Original Message----- From: Histo Jock [mailto:histojock@hotmail.com] Sent: Thursday, May 27, 2004 10:14 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Tissue Arrays Making tissue arrays by re-embedding affects staining "down the road" because of basic chemistry. Each time a specimen is exposed to air there will be some loss of some antigens and nucleic acids due to oxidation from atmospheric ozone, acids, etc circulating in the lab. Not all antigens are affected, not every antibody has a problem with it, but it does happen and is well recognized in tissue array labs. Many labs have documented this phenomenon. Most notable is David Rimm's lab at Yale that has made an art out of preserving antigens on tissue arrays by storing them in a special cabinet filled with nitrogen gas. They see dramatic losses in staining intensity in sections left in air for just a few days. The problem with reheating tissue array cores is that this oxidation effect is grossly accelerated by the higher temperatures and air is allowed to penetrate farther into the core once the paraffin has softened from the heat. Some labs have stopped anealing cores into tissue array blocks at 37 degrees because they see a loss in staining. I have had presonal experience with loss of in-situ signal in blocks annealed at 32 degrees versus ones kept at room temp. I remember at least one study that shows slides stored at higher tempuatures (25 degrees, I think) lose some anitgens in fairly short order. If you haven't seen a problem it's probably because of the antibodies you're using. Many polyclonals will do fine, many monoclonals won't. The more the heat+time+exposure to the atmosphere the more the effect. As with heat+time+everything else in histotechnology it just depends on your specific circumstances. The basic message is that less physical manipulation of a specimen is ALWAYS better than more. There's no reason to heat a specimen to make an array when you can do it perfectly well at room temp. Re-embedded tissue arrays may work great for a lot of things, but experience shows that they do have problems in some applications. HistoJock >Date: Mon, 10 May 2004 04:49:52 +0000 >From: "Thom Jensen" >Subject: Re: [Histonet] Tissue Array >To: Histonet@lists.utsouthwestern.edu >Message-ID: >Content-Type: text/plain > > > How does melting paraffin embedded tissues effect the staining down > the road? That doesn't make since. I have made dozens of multiple > punch arrays by melting the punches and embedding them as you would > normally embed tissues and it has never effected the staining, "DOWN > THE ROAD....." > > Thom >>From: "Histo Jock" >>To: Histonet@lists.utsouthwestern.edu >>Subject: Re: [Histonet] Tissue Array >>Date: Fri, 07 May 2004 20:03:31 -0400 >> >> >>You might want to be careful using Zymed's arrays. I don't think >>they are made with the standard coring method. Rather, I think that >>they use some sort of melting / re-embedding process that can effect >>staining down the road. >> >>HistoJock _________________________________________________________________ Express yourself with the new version of MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> colobio.com Fri May 28 14:26:51 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] anti-CD11b In-Reply-To: <3.0.6.32.20040528095152.00c12830@gemini.msu.montana.edu> Message-ID: Gayle, CD11b works fine on frozen sections but my investigator wanted to use it on mouse liver bile ducts ffpe, I tried vigoursly using the m on m staining system but one of the problems was that it was not frozen to begin with, and another major problem I could not over come was that there is so much endogenous biotin in mouse liver I could not ever block enough to get around non-specific binding (even in frozens) of the biotin. Does anyone sell a m on m IHC staining system that does not use biotinylation? Something like the labelled polymers would be great. I realize that the cd68 was anti-human but it did work on some of the mouse samples. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gayle Callis Sent: Friday, May 28, 2004 9:52 AM To: pengbw@sjtu.edu.cn; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] anti-CD11b Patsy wrote: I have although had some limited success with DAKO CD68 PGM1 clone which only stains macrophages and monocytes and not all the other myleoid cells, but it is not easy and inconsistant in my hands. Caveat: This is a mouse antiHuman CD68 for macrophages, and you will run into the problem of staining with mouse antibody on mouse tissue, a whole other problem introduced requiring mouse on mouse kits and adds to expense of procedure. You can avoid this problem by using CD11b aka Mac1 (Clone M1/70) Rat antiMouse Monoclonal from BD Pharmingen. Do a dilution panel starting at 10 ug/ml, normally you can have good staining at 1:500 - 1:1000 or more depending on the chromogen used, primary has original conc of 0.5mg/ml. and incubated for 30 min at room temperature. Be sure to use the isotype matched control or rat IgG as a negative control We have superb success with acetone(75ml/absolute ethanol (25 ml) fixation. Make sure you air dry the frozen sections OVERNIGHT AT ROOM TEMPERATURE, then fix in this fixative for 5 min at Room temperature, do not dry after fixation but go immediately to 3 changes of PBS (Dulbeccos) and proceed to other blocking steps (DAKO peroxidase block for frozen sections), biotin blocking (Vector), followed by IHC. For a secondary antibody, Goat antiRat bioitinylated, adsorbed to mouse and is an F(ab')2 frag, from Biosource or a Donkey antiRat biotinylated, F(ab')2 frag of IgG adsorbed to Mouse biotinylated from Jackson Immunoresearch is recommended. This secondary should be diluted in the normal serum block containing mouse serum (mouse serum concentration varies from 1 - 5%, we take the middle concentration of 2.5%). We also prefer to use this antibody biotinylated - this eliminates the secondary antibody, and you go directly to Strepavidin-HRP for 20 min. We do not use kits, and our Strepavidin-HRP comes from Biosource (0.5 to 0.6mg/ml diluted 1:500). For a block, we prefer normal serum 10% goat or Donkey (matched to host of secondary) with 2.5% mouse serum (heat inactivated) - Our morphology is excellent, better than acetone fixed frozen sections, and our results are very clean, without any background staining. Make sure you positive control is from an animal stimulated to produce macrophages in order to control the chromogen staining with a microscope. At 09:18 AM 5/28/2004 +0800, you wrote: >Thanks you all, >In fact, I\'m going to do it on frozen sections. >The most prior component I want to stain is monocytes. >So maybe I should check DAKO\'s mAb mentioned by Patsy. > >Baowei Peng _______________________________________________ >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 gcallis <@t> montana.edu Fri May 28 15:16:39 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] anti-CD11b In-Reply-To: References: <3.0.6.32.20040528095152.00c12830@gemini.msu.montana.edu> Message-ID: <3.0.6.32.20040528141639.00c09c48@gemini.msu.montana.edu> Why not come back with a secondary that is directly labeled, and have NO biotin in system at all. Mac1 stains so strongly on frozens(that is what he is doing), he could use goat antiRat-HRP adsorbed to mouse or Donkey antiRat-HRP adsorbed to mouse, F(ab')2 fragments of IgG preferred to avoid fc receptor cross reactivity. Biosource and Jackson Immunoresearch have these antibodies. We have made our own mouse on mouse kits using Scyteks blocking reagents, same as they use in their kits with similar instructions for use, and then a secondary that is not biotinylated. May take more work, but I would do directly labelled secondary first with Rat antiMouse CD11b to avoid biotin altogether in problem liver. One could even use a FITC labelled rat antiMouse CD11b and come back with antiFITC, no biotin involved here either. The secondary rabbit antiFITC from DAKO can be detected by their polymer kit, if I remember correctly. The joy is a primary useful for both IHC and FACS. Chemicon mouse on mouse immunohistochemistry staining kits ARE biotin free - we tried on and had clean results but we DID use frozen sections. At 01:26 PM 5/28/2004 -0600, you wrote: >Gayle, >CD11b works fine on frozen sections but my investigator wanted to use it on >mouse liver bile ducts ffpe, I tried vigoursly using the m on m staining >system but one of the problems was that it was not frozen to begin with, and >another major problem I could not over come was that there is so much >endogenous biotin in mouse liver I could not ever block enough to get around >non-specific binding (even in frozens) of the biotin. Does anyone sell a m >on m IHC staining system that does not use biotinylation? Something like >the labelled polymers would be great. >I realize that the cd68 was anti-human but it did work on some of the mouse >samples. >Patsy > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gayle >Callis >Sent: Friday, May 28, 2004 9:52 AM >To: pengbw@sjtu.edu.cn; Histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] anti-CD11b > > >Patsy wrote: > >I have although had some limited success with DAKO CD68 PGM1 >clone which only stains macrophages and monocytes and not all the other >myleoid cells, but it is not easy and inconsistant in my hands. > >Caveat: This is a mouse antiHuman CD68 for macrophages, and you will run >into the problem of staining with mouse antibody on mouse tissue, a whole >other problem introduced requiring mouse on mouse kits and adds to expense >of procedure. You can avoid this problem by using CD11b aka Mac1 (Clone >M1/70) Rat antiMouse Monoclonal from BD Pharmingen. Do a dilution panel >starting at 10 ug/ml, normally you can have good staining at 1:500 - 1:1000 >or more depending on the chromogen used, primary has original conc of >0.5mg/ml. and incubated for 30 min at room temperature. Be sure to use the >isotype matched control or rat IgG as a negative control We have superb >success with acetone(75ml/absolute ethanol (25 ml) fixation. Make sure you >air dry the frozen sections OVERNIGHT AT ROOM TEMPERATURE, then fix in this >fixative for 5 min at Room temperature, do not dry after fixation but go >immediately to 3 changes of PBS (Dulbeccos) and proceed to other blocking >steps (DAKO peroxidase block for frozen sections), biotin blocking >(Vector), followed by IHC. > >For a secondary antibody, Goat antiRat bioitinylated, adsorbed to mouse and >is an F(ab')2 frag, from Biosource or a Donkey antiRat biotinylated, >F(ab')2 frag of IgG adsorbed to Mouse biotinylated from Jackson >Immunoresearch is recommended. This secondary should be diluted in the >normal serum block containing mouse serum (mouse serum concentration varies >from 1 - 5%, we take the middle concentration of 2.5%). > >We also prefer to use this antibody biotinylated - this eliminates the >secondary antibody, and you go directly to Strepavidin-HRP for 20 min. We >do not use kits, and our Strepavidin-HRP comes from Biosource (0.5 to >0.6mg/ml diluted 1:500). > >For a block, we prefer normal serum 10% goat or Donkey (matched to host of >secondary) with 2.5% mouse serum (heat inactivated) - > >Our morphology is excellent, better than acetone fixed frozen sections, and >our results are very clean, without any background staining. Make sure you >positive control is from an animal stimulated to produce macrophages in >order to control the chromogen staining with a microscope. > > At 09:18 AM 5/28/2004 +0800, you wrote: >>Thanks you all, >>In fact, I\'m going to do it on frozen sections. >>The most prior component I want to stain is monocytes. >>So maybe I should check DAKO\'s mAb mentioned by Patsy. >> >>Baowei Peng >_______________________________________________ >>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 > > 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 carl.hobbs <@t> kcl.ac.uk Fri May 28 17:48:34 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] re caspase 3 and pcna Message-ID: <004401c44505$e8957b00$3c292bd9@home> I understand ( tho I may be corrected and informed further), that PCNA is not the best marker to use, if you are assessing cells in cycle. It was, before Ki67 and H3? --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.690 / Virus Database: 451 - Release Date: 22/05/2004 From dellav <@t> musc.edu Fri May 28 16:18:35 2004 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] Steve Slap Message-ID: Dorothy, I am incredibly grateful for your efforts to keep us informed about Steven's progress. The news particularly hit home for me as I have been a motorcyclist for many years and currently ride every chance I get. Each time I hear of a "rider down" story I take pause and ask myself if it's time to get a new hobby. I have been a biker since I was 19. Steven is someone dear to me in the sense that he and I have collaborated and interacted over many years. He is one of those genuinely nice guys who is always a pleasure to see and speak with. I appreciate his intellect and his reasoned jusdgement. An all around good guy that is one of our NSH family. He collaborated with me by presenting at state and region meetings when I still resided in New York. I never knew that he is a fellow rider so this is something we have not had an opportunity to share. I have prayed for Steven every chance I've gotten. He has been on my mind before I drifted off to sleep since I learned of his accident. I believe in the power of prayer and can only hope that his full recovery is part of God's plan for him. I have so many questions about what happened to cause his accident but of course in the absence of witnesses and his ability to communicate there isn't any way we will know. Bikers often want to learn from these events iin the hope that such knowledge might help them to avoid the very same circumstances. It is often very difficult for non-riders to understand those of us willing to accept the risk of pursuing a pasttime that is inherently dangerous. much like I cannot understand what could possibly compel someone to climb Mt Everest and although I am adventurous and a bit of a risk taker, I will have no problem feeling fulfilled without the Everest experience. Mortorcylcing became incrementally more dangerous as SUV's began taking over the roads. For some reason, perhaps because the driver of these vehicles feels safe inside that huge machine, SUV drivers as a group tend to allow themselves to get distracted and then engage in risky behaviors that can be very harmful to others, like focusing more on their cellphone conversation or applying their makeup. But I digress. As I said, many who've never ridden cannot understand our folly of getting on a bike and riding alongside much larger vehicles at highway speeds for a thrill and sense of freedom that seems so much less important than life itself. Only a biker can understand why a dog will stick his head out of the window of a moving vehicle. While I cannot speak for Steven I suspect that my comments may reflect some of his very own feelings. We riders intellectually know that tragedy may strike but of course we hope that it won't happen to us. we need to think that we are in sufficient control to prevent such an outcome yet in the back of our minds we know that we simply cannot prevent every possible hazard. the bottom line is that we accept the risk in order to pursue our passion. riding provides a joy to us that is hard to verbalize to others. I've told my family that should I lose my life in a cycle accidentl I hope they will take some comfort in knowing that I died doing something I truly loved. Many are much less fortunate. On behalf of NSH, I want you and Lisa to know how much Steven means to all of us. He is truly a member of the NSH family, has given selflessly to share his expertise with others and has been a dear friend and colleague to so many. He has touched many lives and is presently in the thoughts and prayers of I dare say thousands. I know Dorothy that the news of his accident must be particulary difficult for you. We Care! You are family too. sincerely Vinnie Della Speranza >>> 05/28/04 02:36PM >>> So many of you have asked about Steve Slap that I thought the best way to get the word out is through the net. As many of you have already heard, Steve was in accident while riding his Honda scooter. No one saw what actually happened, but thank God he was found quickly. It happened near his home on May 11th and he was airlifted to the trauma center in Springfield. I spoke with Lisa (Steve's significant other) just this morning. Steve is still in a coma and in ICU. For now, it's just a waiting game. The doctor's are optimistic that he will come out of the coma but have given Lisa no time frame. They say he is otherwise healthy and that will work well for the long recovery process. When or if he actually gets back on his feet is not known because there is brain stem damage. If there is any change I will let you all know. The best we can do now is to keep Steve and his family in our prayers. Lisa did say she would love to hear from Steve's friends. So if you would like to send a card, that would be great. Include a picture, if you have one, with who and what it's about written on the back. Part of the healing process is to just keep talking to Steve. Cards and pictures will give Lisa and the nurses different things to talk about with him. Steve's son Jeremy is holding his own. He's a good kid. Their address is: Lisa Smith & Steve Slap PO Box 31 Lake Pleasant, MA 01347 I will pass along any greetings posted to HistoNet. If you prefer, you can email me directly at traczyk7@aol.com Regards, Dorothy Murphy Traczyk Hacker Instruments & Industries Inc. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From maxim_71 <@t> mail.ru Fri May 28 08:05:53 2004 From: maxim_71 <@t> mail.ru (=?koi8-r?Q?=22=F0=C5=DB=CB=CF=D7=20=ED=C1=CB=D3=C9=CD=22=20?=) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] exotic species/chief&parietal cells Message-ID: Sharon, I use staining with Congo red (Highman) for parietal cells. Granules of the chief cells stains in dark blue color with Giemsa techniques (or his modifycations). Maxim Peshkov, Pathoanatomical bureau, Taganrog, Russia. Maxim_71@mail.ru From KHays <@t> mbhs.org Thu May 27 15:32:29 2004 From: KHays <@t> mbhs.org (KHays@mbhs.org) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] steve slapp Message-ID: Kathy Tedford-Hays HT (ASCP) Technical Specialist, Histology Dept (601)-968-3070 ext 7398 Baptist Medical Center 1225 North State Street Jackson, MS 39202 i would like to ask everyone getting this message to take time out of your busy day and pray for steve slapp. although i have never met him personally, he has replied to many of our questions over the last year and many of you know him and care deeply for him. join me now in praying for a full recovery. the power of prayer is amazing. MATTHEW 21 VERSE 22. From JWEEMS <@t> sjha.org Sat May 29 03:11:25 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] Unsubscribe Message-ID: 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 David.Edmondson <@t> christie-tr.nwest.nhs.uk Sat May 29 05:10:54 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] HYALURONIDASE Message-ID: Hi,all, uronidase (EC3.2.1.35) Sigma Cat H-3206 has been doing fine for us Dave Christie's Manchester UK -----Original Message----- From: Mcleod [mailto:hmcleod@chempath.uct.ac.za] Sent: 28 May 2004 14:48 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HYALURONIDASE Dear All We need to order hyaluronidase. We currently are using a MERCK PRODUCT (K 27072808) which (acording to our local agency) is no longer available from Merck. Whose product do histonetters recommend .... ...and which of the Sigma products would you recommend? Thankyou for all responses Heather _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From David.Edmondson <@t> christie-tr.nwest.nhs.uk Sat May 29 05:12:46 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] anti-CD11b Message-ID: Envision kit? Dave -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: 28 May 2004 21:17 To: Patsy Ruegg; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] anti-CD11b Why not come back with a secondary that is directly labeled, and have NO biotin in system at all. Mac1 stains so strongly on frozens(that is what he is doing), he could use goat antiRat-HRP adsorbed to mouse or Donkey antiRat-HRP adsorbed to mouse, F(ab')2 fragments of IgG preferred to avoid fc receptor cross reactivity. Biosource and Jackson Immunoresearch have these antibodies. We have made our own mouse on mouse kits using Scyteks blocking reagents, same as they use in their kits with similar instructions for use, and then a secondary that is not biotinylated. May take more work, but I would do directly labelled secondary first with Rat antiMouse CD11b to avoid biotin altogether in problem liver. One could even use a FITC labelled rat antiMouse CD11b and come back with antiFITC, no biotin involved here either. The secondary rabbit antiFITC from DAKO can be detected by their polymer kit, if I remember correctly. The joy is a primary useful for both IHC and FACS. Chemicon mouse on mouse immunohistochemistry staining kits ARE biotin free - we tried on and had clean results but we DID use frozen sections. At 01:26 PM 5/28/2004 -0600, you wrote: >Gayle, >CD11b works fine on frozen sections but my investigator wanted to use it on >mouse liver bile ducts ffpe, I tried vigoursly using the m on m staining >system but one of the problems was that it was not frozen to begin with, and >another major problem I could not over come was that there is so much >endogenous biotin in mouse liver I could not ever block enough to get around >non-specific binding (even in frozens) of the biotin. Does anyone sell a m >on m IHC staining system that does not use biotinylation? Something like >the labelled polymers would be great. >I realize that the cd68 was anti-human but it did work on some of the mouse >samples. >Patsy > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gayle >Callis >Sent: Friday, May 28, 2004 9:52 AM >To: pengbw@sjtu.edu.cn; Histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] anti-CD11b > > >Patsy wrote: > >I have although had some limited success with DAKO CD68 PGM1 >clone which only stains macrophages and monocytes and not all the other >myleoid cells, but it is not easy and inconsistant in my hands. > >Caveat: This is a mouse antiHuman CD68 for macrophages, and you will run >into the problem of staining with mouse antibody on mouse tissue, a whole >other problem introduced requiring mouse on mouse kits and adds to expense >of procedure. You can avoid this problem by using CD11b aka Mac1 (Clone >M1/70) Rat antiMouse Monoclonal from BD Pharmingen. Do a dilution panel >starting at 10 ug/ml, normally you can have good staining at 1:500 - 1:1000 >or more depending on the chromogen used, primary has original conc of >0.5mg/ml. and incubated for 30 min at room temperature. Be sure to use the >isotype matched control or rat IgG as a negative control We have superb >success with acetone(75ml/absolute ethanol (25 ml) fixation. Make sure you >air dry the frozen sections OVERNIGHT AT ROOM TEMPERATURE, then fix in this >fixative for 5 min at Room temperature, do not dry after fixation but go >immediately to 3 changes of PBS (Dulbeccos) and proceed to other blocking >steps (DAKO peroxidase block for frozen sections), biotin blocking >(Vector), followed by IHC. > >For a secondary antibody, Goat antiRat bioitinylated, adsorbed to mouse and >is an F(ab')2 frag, from Biosource or a Donkey antiRat biotinylated, >F(ab')2 frag of IgG adsorbed to Mouse biotinylated from Jackson >Immunoresearch is recommended. This secondary should be diluted in the >normal serum block containing mouse serum (mouse serum concentration varies >from 1 - 5%, we take the middle concentration of 2.5%). > >We also prefer to use this antibody biotinylated - this eliminates the >secondary antibody, and you go directly to Strepavidin-HRP for 20 min. We >do not use kits, and our Strepavidin-HRP comes from Biosource (0.5 to >0.6mg/ml diluted 1:500). > >For a block, we prefer normal serum 10% goat or Donkey (matched to host of >secondary) with 2.5% mouse serum (heat inactivated) - > >Our morphology is excellent, better than acetone fixed frozen sections, and >our results are very clean, without any background staining. Make sure you >positive control is from an animal stimulated to produce macrophages in >order to control the chromogen staining with a microscope. > > At 09:18 AM 5/28/2004 +0800, you wrote: >>Thanks you all, >>In fact, I\'m going to do it on frozen sections. >>The most prior component I want to stain is monocytes. >>So maybe I should check DAKO\'s mAb mentioned by Patsy. >> >>Baowei Peng >_______________________________________________ >>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 > > 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 FARFAT <@t> aol.com Sat May 29 23:03:47 2004 From: FARFAT <@t> aol.com (FARFAT@aol.com) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] unsuscribe Message-ID: Please unsuscribe as I am going on vacation for 1 month starting june 2004 Akbar. From raghul <@t> sctimst.ker.nic.in Sun May 30 23:09:18 2004 From: raghul <@t> sctimst.ker.nic.in (Dr. Raghul) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] IHC_swine vessels_cross reactivity Message-ID: Dear all, I need to identify macrophages and endothelial cells in resin embedded pig blood vessels with stent. I couldnt find any specific primary antibodies against swine macrophages or endothelial cells and has to rely on cross reactivity pirmarily intended in human tissue. Has any one tried such cross reactive antibodies in swine tissue or is there specific ones against pig which ive missed. Please help J. Raghul, Scientist -C, sri chithra tirunal institute of medical sciences, Trivandrum , India From psanquin <@t> lugo.usc.es Mon May 31 03:52:28 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] Sucrose embedding In-Reply-To: <000001c43e85$a96900e0$0a4dbad0@hppav> Message-ID: <3.0.6.32.20040531105228.007bd100@pop.lugo.usc.es> A long time has gone since my last immunos in criostat cut sections. I have forgotten a simple question. When the sucrose embedding of the tissues must be done? Should be made just the night before cutting or just after the fixation is finished? Thanks in advance Pablo Sanchez-Quinteiro From Inga.Hansson <@t> neuro.uu.se Mon May 31 07:44:36 2004 From: Inga.Hansson <@t> neuro.uu.se (Inga Hansson) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] methyl green Message-ID: Hi histonetters! Can anyone explain to me why my methyl green counterstaining stopped working?? I use 0.5% solution and have had it for a while. Should it be made fresh?? Spinal cord seems to be the least stained! Cerebellum the best. Is there a difference between different cell types?? Thanks in advance! Inga From hmcleod <@t> chempath.uct.ac.za Mon May 31 07:44:36 2004 From: hmcleod <@t> chempath.uct.ac.za (Mcleod) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] p53 Message-ID: <40BB28B4.5812F34@chempath.uct.ac.za> Hi All We need to stain for p53 on ffpe murine git material. Would the Novocastra NCL-p53-CM5p be one of the better antibodies to use? Has anyone used the NCL-p53-505? Any answers and advice would be very welcome. Also thanks for all the responses the hyaluronidase question. Heather From ctsblack <@t> capeheart.uct.ac.za Mon May 31 08:18:22 2004 From: ctsblack <@t> capeheart.uct.ac.za (Melanie Black) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] Visualizing GFP-Thanks! Message-ID: Fellow Colleagues Many Thanks to all of you who responded with such helpfull information on GFP!! This really is the most useful laboratory assistant!!!! Specially to Nancy and Daniel! Thanks Again 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 nyilmaz <@t> mersin.edu.tr Mon May 31 08:39:27 2004 From: nyilmaz <@t> mersin.edu.tr (=?windows-1254?Q?Nejat_Y=FDlmaz?=) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] Is LEO 906 still produced??? Message-ID: <001601c44714$b1aa9100$4e01a8c0@mersin.edu.tr> Hello Everybody... We are planning to buy a TEM. We want to learn whether LEO 906 is still produced by Zeiss. Because we couldn't find any information about LEO 906 in their official web page. However the dealer of Zeiss in Turkey told us it is still being produced by Zeiss. We couldn't trust him since he told us LEO 906 is not produced any more in our previous conversation with him. If anybody help us about this problem we would be very happy. Thanks in advance... Dr. Necat Y?lmaz From Ckiely128 <@t> aol.com Mon May 31 10:23:29 2004 From: Ckiely128 <@t> aol.com (Ckiely128@aol.com) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] unsubscribe, thank you Message-ID: <155.362ea2d4.2deca7f1@aol.com> From ylee <@t> bccancer.bc.ca Mon May 31 13:57:40 2004 From: ylee <@t> bccancer.bc.ca (Yin Ping Lee) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] Rabbit anti-FITC-HRP conjugate Message-ID: <6BAF4D075F07D411B30900508B94CBA0C1F564@SERVER20> Hi, Histonetter: Does anyone have experience with rabbit anti-FITC-HRP conjugate from Dako? Does it work not as well and what is the problem exactly? I am using this product for the EBER ISH staining and have occasionally encountered mysterious background staining. Yin Ping Lee Histopathology Lab BC Cancer Agency Vancouver, BC From mivalmar <@t> yahoo.es Mon May 31 16:09:55 2004 From: mivalmar <@t> yahoo.es (=?iso-8859-1?q?Miguel=20Valenzuela?=) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] VEGF problems in Immunohistochemistry Message-ID: <20040531210955.52279.qmail@web12609.mail.yahoo.com> Hi all I need to find a paper to know what kind of cells are stained with VEGF (Vascular endotelial growth factor)... Yes, I know that endothelial cells are stained but I get also stain in adipose cells. I want to learn more about this antibody and others (tie, angiostatin, VEGF-Receptors 1,2) Any help will be useful. Thanks very much. Miguel Valenzuela. Spain. __________________________________________________ Correo Yahoo! - 6 MB, antivirus y antispam ?gratis! Reg?strate ya - http://correo.yahoo.es From mivalmar <@t> yahoo.es Mon May 31 16:12:02 2004 From: mivalmar <@t> yahoo.es (=?iso-8859-1?q?Miguel=20Valenzuela?=) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] apoptosis protocol (anexin, caspase) for immunohistochemistry Message-ID: <20040531211202.87675.qmail@web12607.mail.yahoo.com> Hi all I would like to contact someone that has used anexin-V and/or caspase-3 in paraffin tissues. I have a lot of nuclear stain with tunel. And I would like to find a protocol. Any help will be useful Thanks. Miguel V-S. Spain. ______________________________________________________________________ Correo Yahoo! - 6MB, m?s protecci?n contra el spam ?Gratis! http://correo.yahoo.es From int09018 <@t> alphahunt.com Mon May 31 18:56:02 2004 From: int09018 <@t> alphahunt.com (HCS) Date: Fri Sep 16 15:23:03 2005 Subject: [Histonet] Older embedding center in ebay. Message-ID: <000b01c4476a$d55f13e0$6501a8c0@hp> Hi, I have listed a spare embedding center that I no longer need on ebay in case anyone would be interested in getting one. The item number is 3818030988. It is a Reichert-Jung. LeRoy Brown HT(ASCP) HTL Histology Consultation Services 1-360-966-7300 www.histocs.com Everson, Washington.