From docmichel <@t> netbulmail.com Wed Oct 1 06:13:25 2003 From: docmichel <@t> netbulmail.com (izzet oguz) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Rhodizonate for lead Message-ID: <3640.193.255.128.130.1065006805.webmail@mail1.netbulmail.com> Hi there, we are trying to demonstrate lead with rhodizonate method..but it hasn't worked yet..here is our method from "Theory and Practice of Histological Techniques" : prep of solution : sodium rhodizonate 200 mg distilled water 99 cm3 glacial acetic acid 1 cm3 ( FIRST DISTILLED WATER+GAA THAN ADDED 200 MG RHODIZONATE) Method : 1)sections to water 2)transfer to rhodizonate sol- 1 hour 3)wash in tap water 4)counterstain in 0.1 percent light green in 1 percent acetic acid- 2 minutes (I THINK THE RAT?O 1:1, I MEAN 100 CC %0.1 L?GHT GREEN + 100 CC % 1 ACETIC ACID ?) 5)rinse briefly in tap water 6)mount in glycerin jelly (WE D?D TH?S W?TH UNFORTUNATELY WITH CANADA BALSAM; DOES ?T AFFECT THE RESULT?) I would like to learn every kind of suggestion from you..and also experiences.. Thanks in advance.. ?zzet From Noel.Clark <@t> ORTHO.UAB.EDU Wed Oct 1 07:45:10 2003 From: Noel.Clark <@t> ORTHO.UAB.EDU (Noel D. Clark) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Perfusion techniques in mice/rats Message-ID: <85F6C7A1330E794DB8540AFD001CC77E0D670F@ROSCO> Was hoping to get input on the various perfusion techniques used by other research labs for rodents. As always, any help is greatly appreciated. See you in Louisville! Thanks in advance, Noel Noel Clark, M.A., HTL (ASCP) 1919 7th Ave South Orthopaedic Research Laboratory Center for Metabolic Bone Disease University of Alabama at Birmingham Birmingham, Alabama 35294 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/bfb24562/attachment.htm From decorah <@t> rarc.wisc.edu Wed Oct 1 08:27:59 2003 From: decorah <@t> rarc.wisc.edu (Maureen Decorah) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] OCT Buffer for histo and cryopreservation Message-ID: My pathologist wanted me to find out if any one uses this technique and what sort of costs would be associated with it. Thank you Maureen > >| From: >| Date: Thu, 25 Sep 2003 11:10:53 -0500 >| Subject: formalin >| >| >| >| Greetings BBAG members, >| >| I am writing in response to postings last month concerning formalin >| collections: >| >| Modern methods of research and diagnostics are increasingly turning >| to the molecular genetic level. Thus it is becoming increasingly >| important for those dealing with any aspect of tissue sampling to be >| aware of the potential downstream utility of the specimens that they >| collect. Simply maintaining jars of formalin-fixed material is >| difficult to justify in light of storage space and maintenance costs >| compared with the lack of downstream utility of the specimen for >| molecular analysis (formalin causes DNA to bind to histone proteins, >| making it useless for all but the crudest of molecular assays and it >| virtually destroys RNA). >| >| What many research hospitals and biotech companies conducting >| pathology research prefer to do is: collect the tissue specimen in >| OCT buffer and cryopreserve it. Such a sample is amenable to both >| traditional sectioning as well as DNA/RNA analysis - which helps >| justify collecting and retaining the sample since it has value for >| other as yet unidentified research purposes. It is important to >| realize that such a sample takes up far less space than a jar of >| formalin, and assuming the freezer doesnt malfunction, requires less >| maintenance as well. Cryopreserved tissues can be used in molecular >| assays ranging from paternity exclusion, to retrospective >| epidemiological surveys and environmental toxicology examinations. >| Such a collection is of far greater prospective value than >| traditional collections and individuals/institutions need to >| carefully examine the underlying justification for making biomaterial >| collections in light of limited financial resources. A plastic >| cryovial is far less expensive than a jar of formalin - and OCT >| buffer is much less toxic than formalin. >| >| It is easy to make a strong case for archiving frozen tissues, >| despite the up front capital costs for freezers... Ultimately, using >| liquid Nitrogen protects your collection from mechanical freezer >| failure and power outages, as well as providing temperatures cold >| enough to preserve viable cells, including gametes and embryos. Thus, >| Many institutions can benefit from establishing a centralized >| biomaterial core facility which can serve many different researchers >| - from vets and pathologists, to conservation geneticists and repro >| specialists. It can also be advantageous to enter into a strategic >| partnership with a dedicated cryostorage facility which can backup >| your collection at a remote fail-safe location. >| >| No matter what you collect or how you collect it, I cannot overstate >| the importance of having an electronic inventory of your collection >| for rapid information dissemination and specimen retrieval. A >| collection only has value when researchers are aware of its existence >| and contents. Fortunately, there are a number of prepackaged >| software/hardware systems which will allow for computer generation of >| specimen labels, sample tracking, etc. >| >| The international Society for Biological and Environmental >| Repositories (ISBER) will have "Repository Design and Data >| Management" as a theme for our next annual meeting, to be held at the >| American Museum of Natural History May 11-14, 2004. Those interested >| in learning more about modern biomaterial collections and their >| management should consider attending this conference. In the mean >| time, feel free to contact me if you have any specific questions and >| I will do my best to steer to someone who can help answer them (if I >| cannot do so myself). >| >| Robert Hanner, Ph.D. >| Scientific Program Director >| Coriell Institute for Medical Research >| 403 Haddon Avenue >| Camden, NJ 08103 >| >| Voice: (856) 757-9727 >| Fax: (856) 757-9737 >| http://cimr.umdnj.edu >| >| President >| International Society for Biological and Environmental Repositories >| http://www.ISBER.org >| >| > >| >| With respect to Scott's question, "wet" formalin-fixed tissues have been >| >| placed in glass jars with screw top lids for long-term storage. At the >| >| present time, these tissues are being kept permanantly. Full sets of >| >| tissue are trimmed in, paraffin embedded, and a set of glass slides is >| >| prepared for the majority of the collection animals. The blocks and >| >| glass slides are also permanently kept. All of the materials are >| >| presently housed on-site either in a moderately sized room off our >| >| histology lab or in a large metal "container" behind the Health Center. >| >| What happens when all the room is filled? Not yet sure. It is likely >| >| that over the next little while (year or two) we will have to rethink >| >| our policy of keeping everything forever since space issues and several >| >| other problems that John detailed quite nicely (maintenance of the jars >| >| - topping them off, clean-up, and personnel time to manage the archived >| >| tissues) are and will continue to be problems. It is likely that blocks >| >| and slides will be kept permanently and that decisions will have to be >| >| made about the formalin fixed tissues. I can contact you at a later >| >| date once we have decided what our course for the future will be. >| >| >| >| Regarding the usefulness of tissues for molecular diagnostic techniques >| >| after long-term formalin fixation...I have little expertise in this >| >| area, however I seem to have heard or read similar information to what >| >| John describes and have also gotten mixed results when I have run immuno >| >| on tissues that have been stored for a long time in formalin before >| >| being paraffin embedded. >| >| >| >| D >| >| >| >| D McAloose, VMD, Diplomate ACVP >| >| Acting Head, Department of Pathology >| >| Wildlife Conservation Society >| >| Department of Pathology >| >| 2300 Southern Blvd >| >| Bronx, NY 10460 >| >| (718) 220-7105 >| >| dmcaloose@wcs.org >| >| >| >| >| >| > >| >| >| >| ---------------------------------------------------------------- >| >| | From: >| >| | Date: Wed, 30 Jul 2003 07:01:08 -0500 >| >| | Subject: Wet tissue archiving >| >| | >| >| | >| >| | [attachment below: attachment 279f2c-2] >| >| | >| >| | Hello all, >| >| | >| >| | I am trying to get a feel for methods of long-term storage of "wet" >| >| | formalin fixed tissues. I currently use a heat sealed plastic bag >| >| | system in my lab, but am looking for other opinions concerning these >| >| | tissues. Answers or discussion about any or all of these questions >| >| | would be great: >| >| | >| >| | 1. Do you think long term stored wet tissues have value (other than >| >| | histopathology)? >| >| | I was under the general impression that tissues that sit in formalin >| >| | for years are no longer good for genetics, immuno, in situ, PCR, etc >| >| | 2. What is the best way to keep wet tissues long term (10,20,50 years >| >| or >| >| | more)? >| >| | 3. Have you had good or bad experience with plastic bags or any other >| >| | system? >| >| | 4. What happens when you run out of space? >| >| | 5. Some institutions actually trim all of their wet tissues into >| >| | paraffin blocks for storage...does anyone have experience with this? >| >| | 6. Any input would be welcomed >| >| | >| >| | Scott P. Terrell, DVM, Diplomate ACVP >| >| | >| >| | Veterinary Pathologist, Walt Disney World Animal Programs >| >| | Assistant Professor, Pathobiology, University of Florida College of >| >| | Veterinary Medicine >| >| | >| >| | >| >| | Office: (407)938-2746 >| >| | Fax: (407): 938-1909 >| >| | >| >| | Address: >| >| | Veterinary Services, Disney's Animal Kingdom >| >| | 1200 N. Savannah Cir >| >| | Bay Lake, FL 32830 >| >| | >| >| | >| >| | >| >| >| >| >| -- >| > > >-- >Victoria L. Clyde, DVM >Staff Veterinarian >Milwaukee County Zoo >10001 West Blue Mound Road >Milwaukee, WI 53226 >414-771-3040 zoo >414-256-5441 hospital >414-256-5451 office >414-256-2522 fax >vclyde@execpc.com -- @@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@ Maureen Decorah (608) 262-0933 1710 University Avenue (608) 265-2698 Fax 385 Enzyme Institute Madison, WI 53726-4087 decorah@rarc.wisc.edu @@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@ -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/1e30e86c/attachment.htm From settembr <@t> umdnj.edu Wed Oct 1 08:35:35 2003 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Kappa, Lambda staining??? Message-ID: Jesus, I don't know about the Ventana but I do advise you to try diluting your antibodies. Dana Settembre University Hospital-UMDNJ, Newerk, NJ >>> Jesus Ellin 9/30/2003 5:00:18 PM >>> Hello everyone ,, Need some help here, currently our lab is trying to create a IHC protocol for kappa and lambda on the Venntana benchmark,, we are having some real problems with the background staining in our control and patients,, we are using a blocker and or hydrogen peroxide, to try and minimize the peroxidase but we are still getting alot of background, if any one out there that has a benchmark and has a protocol for these stains some help would be appreciated,, forgot to mention that we are using Tonsil and Bone Marrow for control, One other thing how many out there are using the Tamtron System,, we are currently in the building phase and would like to ask some people out there questions concerning proccess, codes and end user training Jesus Ellin HTL ASCP Yuma Regional Medical Center jellin@yumaregional.org From mwhitesi <@t> adventisthealthcare.com Wed Oct 1 08:56:53 2003 From: mwhitesi <@t> adventisthealthcare.com (Marnie Whiteside) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Histology Position in Maryland Message-ID: We currently have a position available doing routine histology work. Experience is special stain and cutting is required. We are located in Takoma Park Maryland (just outside of Washington DC). Any interested applicants can call Marnie Whiteside for further details. Marnie Whiteside Interim Histology Supervisor Washington Adventist Hospital -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/2acbaeef/attachment.htm From kspencer <@t> utmem.edu Wed Oct 1 09:25:36 2003 From: kspencer <@t> utmem.edu (Kathleen Spencer) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Perfusion techniques in mice/rats In-Reply-To: <85F6C7A1330E794DB8540AFD001CC77E0D670F@ROSCO> Message-ID: <1FD2D7DC-F41B-11D7-BAB9-000393967904@utmem.edu> Attached is the cardiac perfusion protocol that we use regularly. Kathleen Spencer HT, ASCP UTHSC Memphis, TN On Wednesday, October 1, 2003, at 07:45 AM, Noel D. Clark wrote: > Was hoping to get input on the various perfusion techniques used by > other research labs for rodents.? As always, any help is greatly > appreciated.? See you inLouisville! > > ? > > Thanks in advance, > > Noel > > ? > > ? > > ? > > ? > > Noel Clark, M.A., HTL (ASCP) > > 1919 7th Ave South > > Orthopaedic Research Laboratory > > Center for Metabolic Bone Disease > > UniversityofAlabamaatBirmingham > > Birmingham,Alabama35294 > > ? > -------------- next part -------------- Skipped content of type multipart/mixed From siksik <@t> vgernet.net Wed Oct 1 09:27:54 2003 From: siksik <@t> vgernet.net (Steven Slap) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] over and under processing In-Reply-To: <20030930170000.1962.52333.Mailman@swlx167.swmed.edu> Message-ID: Hi HistoNetters Louise Renton wrote: "However, anecdotal evidence has stated that these tissues may become 'alcohol-fixed" thus making them difficult to cut. What is the rationale behind this?" In my experience, this is caused by under fixation. Formalin is a very slow acting fixative (it penetrates at a rate of something like .7mm/hour, then still needs to cross-link the proteins), so, in many cases, tissues are not totally fixed when they reach the alcohol steps in processing and are fixed by the alcohols, leaving distinctive alcohol-fixation artifacts (shrinkage, dryness, etc). Therefore, what many people tend to call "over-processing" turns out to be under-fixation. best regards, Steven Slap From gareth.davis <@t> Vanderbilt.Edu Wed Oct 1 09:31:30 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Perfusion techniques in mice/rats Message-ID: <082C721AF78DB34983E8BA2CD085462105C695@mailbe07> Perfusion Protocol Preparation for Perfusion: Fixative =4% PFA Prepare fresh on day of use or the night before and keep at 4oC. 4% Paraformaldehyde 3.8% Sodium Borate Add 40g of paraformaldehyde to 800ml of water and heat to 60oC while stirring. Add 38g of sodium borate, paraformaldehyde will not dissolve until it is added. Cool off to 4oC. Adjust pH to 9.5 with glacial acetic acid. 4% PFA with 10% sucrose= In a 100ml cylinder add 10g of Sucrose, than add 4% PFA to 100ml Perfusion 1. Anesthetize animal, open thoracic cavity, expose heart and visualize ascending aorta. Insert 18-gauge needle (on vacutainer collection tubing) through left ventricle into ascending aorta- if possible, hard to tell sometimes. Clamp cannula into place with would clip or small clamp and then snip right atrium. (I find it's easier - on mice- to hold needle in place, because clamp gets in the way). 2. Slowly perfuse animal with about 40-50ml room temperature saline (it's recommended to use a pump here, but I use 50 ml syringe, and perfuse slowly) until fluid is clear. 3. Change to cold 4% PFA and slowly perfuse animal. When the animal?s forelimbs are fully extended reduce perfusion speed, use about 50ml of fixative (for a 30g mouse). 4. After perfusion is complete, decapitate the animal and quickly remove the brain using a caudal approach. Place brain in 4% PFA with 10% sucrose at 4oC for overnight. (The 4%PFA with sucrose will post-fix brain and cryoprotect tissue at the same time.) Some recommend to post-fix in 4% PFA-only, overnight, then cryoprotect with 30% sucrose for another 24 hours. Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/4b8f1ec0/attachment.htm From cwscouten <@t> myneurolab.com Wed Oct 1 09:33:51 2003 From: cwscouten <@t> myneurolab.com (Charles W. Scouten, Ph.D.) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Perfusion techniques in mice/rats Message-ID: What tissue are you after? For brain, I used a modified version of the EM protocol published by Cragg, 1980. He found a way to avoid shrinkage due to formalin fixation. When the fixative arrives, the sodium pump proteins on the outside of the cell are the first to be fixed. Sodium rushes into the cell down the concentration gradient, and water rushes in to maintain tonicity. The cell swells 30%, and fixation occurs with crosslinking to proteins in neighboring cells. Later, when equilibrium is restored and the cells shrink back, they drag their neighbors back with them. The result is 20% gross shrinkage of the brain, and loss of extracellular space. Cragg reasoned that if you could replace the extracellular fluid with isotonic fluid without sodium, the shrinkage would not occur at all. Isotonic sucrose (close to 5%) fills the bill, but will not replace extracellular fluid because of the blood brain barrier. This can be overcome if the sucrose is pumped through at 300 mm Hg to overcome the blood brain barrier without breaking blood vessels. We offer a suitable apparatus to implement this plan, see the link below, and a protocol. It has the salutary effect of completely and thoroughly washing out the red blood cells. You use sucrose for the prewash instead of saline, very briefly because of the high pressure and flow rate, then get the fixative to the cells more rapidly than traditional methods. Gravity flow is too weak, 150 mm Hg translates to 6.7 ft of water. In your lab space, can the water reservoir be 6.7 ft above the animal on the sink, without raising the ceiling? Or to implement the Cragg perfusion, you would need 13.4 ft. from animal to sucrose. Peristaltic pumps control flow rate, which does not correct for vascular resistance of the animal, and the correct flow rate is not obvious. Try the Perfusion One: http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=4 71001&catdesc=Histology+Equipment&CatThreeID=674&CatOneID=4&subcatdesc=S acrifice+Equipment&idsubcategory=21 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 www.myneurolab.com -----Original Message----- From: Noel D. Clark [mailto:Noel.Clark@ORTHO.UAB.EDU] Sent: Wednesday, October 01, 2003 7:45 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Perfusion techniques in mice/rats Was hoping to get input on the various perfusion techniques used by other research labs for rodents. As always, any help is greatly appreciated. See you in Louisville! Thanks in advance, Noel Noel Clark, M.A., HTL (ASCP) 1919 7th Ave South Orthopaedic Research Laboratory Center for Metabolic Bone Disease University of Alabama at Birmingham Birmingham, Alabama 35294 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/cb8e7fc2/attachment.htm From histo <@t> skm.org.pk Wed Oct 1 09:38:18 2003 From: histo <@t> skm.org.pk (Histology) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] SAKURA TISSUE -TEK SCA Coverslipper Message-ID: > Dear Histonetters, > I am considering purchasing the SAKURA TISSUE -TEK SCA Coverslipper. > I'd appreciate the hearing about your experiences. > Thanks in advanced, > Muhammad Tahseen Histology Supervisor Deptt. Pathology Shaukat Khanum Cancer Hospital And Research Center Lahore. Pakistan Ph. +92 42 5180725-36 Ext 2369, Fax. +92 42 5180723 e-mail. Histology@skm.org.pk From DDittus787 <@t> aol.com Wed Oct 1 09:39:12 2003 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Kappa, Lambda staining??? Message-ID: <40B41256.3E347C88.0A1F969F@aol.com> BONE MARROW WILL GIVE HIGH BACKGROUND DUE TO BLOOD , HOWEVER I WILL TELL YOU THAT ON BOTH THE NEXES AND BENCHMARK WE ONLY INCUBATE 4 MINUTES ON BOTH THE KAPPA AND LAMBDA AND GET NICE RESULTS, YOU COULD TRY DILUTING YOUR ANTIBODY OUT OR YOU CAN LOWER INCUBATION TIMES IN 2-4 MIN INCREMENTS TO SEE IF THIS LOWERS BACKGROUND. DANA From DDittus787 <@t> aol.com Wed Oct 1 09:42:28 2003 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Fw: Clones (ER) Message-ID: <17644810.4AF878BA.0A1F969F@aol.com> Lena: we use clone 6f11, however others swear by 1d5 for er. dana From juan.gutierrez <@t> christushealth.org Wed Oct 1 09:49:03 2003 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Modified Mowry Message-ID: I would also like a copy, Thanks. Juan C. Gutierrez, HT(ASCP) -----Original Message----- From: Jeannie Heck [mailto:jeannie_heck@yahoo.com] Sent: Tue 9/30/2003 3:56 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Modified Mowry I would deeply appreciate a procedure for a modified Mowry Colloidal Iron using 3% as a mordant before the colloidal iron stain. I am currently using the Muller Mowry and I am getting inconsistent results. All help is appreciated. Please post to this list or e-mail me directly at heckj@hhosp.com. Thanks in advance. Jeannie Heck HT (ASCP) _____ Do you Yahoo!? The New Yahoo! Shopping - with improved product search From marirose.satterfield <@t> MercyMemorial.org Wed Oct 1 09:58:02 2003 From: marirose.satterfield <@t> MercyMemorial.org (Satterfield, Marirose) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] hardware and foreign bodies Message-ID: Our surgical doctors want to be able to give their patients their hardware and FB without it being grossed in the Histology Lab. I guess they say it would save the patient money by not having a pathology fee. I was wondering how other hospital handle their hardware and FB. Thanks in advance Mari Mercy Memorial Hospital From juan.gutierrez <@t> christushealth.org Wed Oct 1 10:06:23 2003 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Kappa, Lambda staining??? Message-ID: That's just the nature of the beast. Kappa and Lambda are known for excesive background staining. My sugestion: since you already have the Benchmark, get the probes. I ran the probes and the IHC together this summer and the difference in results were amazing. There is no background and more cells staining with the probe. I'm currently trying to convince my manager to switch us to probes without any luck so far. In the meantime we are using an Fc Receptor Blocker from INNOVEX Biosciences as an option on our program. It helps, but we still get some background. Juan C. Gutierrez HT(ASCP) -----Original Message----- From: Jesus Ellin [mailto:Ellin13@adelphia.net] Sent: Tue 9/30/2003 7:00 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Kappa, Lambda staining??? Hello everyone ,, Need some help here, currently our lab is trying to create a IHC protocol for kappa and lambda on the Venntana benchmark,, we are having some real problems with the background staining in our control and patients,, we are using a blocker and or hydrogen peroxide, to try and minimize the peroxidase but we are still getting alot of background, if any one out there that has a benchmark and has a protocol for these stains some help would be appreciated,, forgot to mention that we are using Tonsil and Bone Marrow for control, One other thing how many out there are using the Tamtron System,, we are currently in the building phase and would like to ask some people out there questions concerning proccess, codes and end user training Jesus Ellin HTL ASCP Yuma Regional Medical Center jellin@yumaregional.org From JGordon <@t> cellmarque.com Wed Oct 1 10:05:32 2003 From: JGordon <@t> cellmarque.com (Jeff Gordon) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Kappa, Lambda staining??? Message-ID: Jesus, I can't recall a single lab that we introduced our MONOCLONAL kappa and lambda to that didn't solve their background problem, especially with bone marrows. We have plenty of clients that use our monoclonal kappa and lambda on a variety of stainers, and we have found that often times heat retrieval with EDTA gives cleaner results than enzyme retrieval. I can supply you with references if needed. http://www.cellmarque.com/2000/Pages/catalog/kappa.html http://www.cellmarque.com/2000/Pages/catalog/lambda.html Jeff Gordon Domestic Technical Sales and Marketing Manager Cell Marque Corp. 1-800-665-7284, Ext. 12 -----Original Message----- From: Jesus Ellin [mailto:Ellin13@adelphia.net] Sent: Tuesday, September 30, 2003 7:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Kappa, Lambda staining??? Hello everyone ,, Need some help here, currently our lab is trying to create a IHC protocol for kappa and lambda on the Venntana benchmark,, we are having some real problems with the background staining in our control and patients,, we are using a blocker and or hydrogen peroxide, to try and minimize the peroxidase but we are still getting alot of background, if any one out there that has a benchmark and has a protocol for these stains some help would be appreciated,, forgot to mention that we are using Tonsil and Bone Marrow for control, One other thing how many out there are using the Tamtron System,, we are currently in the building phase and would like to ask some people out there questions concerning proccess, codes and end user training Jesus Ellin HTL ASCP Yuma Regional Medical Center jellin@yumaregional.org -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/dfee7ba8/attachment.htm From juan.gutierrez <@t> christushealth.org Wed Oct 1 10:11:52 2003 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Re: Modified Mowry (Long discussion) Message-ID: I believe she's referring to 3% Acetic acid. -----Original Message----- From: John Kiernan [mailto:jkiernan@uwo.ca] Sent: Tue 9/30/2003 11:32 PM To: Jeannie Heck; Histonet Listserver Cc: Subject: [Histonet] Re: Modified Mowry (Long discussion) gJeannie Heck wrote: > > I would deeply appreciate a procedure for a modified > Mowry Colloidal Iron using 3% as a mordant before the > colloidal iron stain. I am currently using the Muller > Mowry and I am getting inconsistent results. All help > is appreciated. Please post to this list or e-mail me > directly at heckj@hhosp.com. Thanks in advance. > Jeannie Heck HT (ASCP) ___________________________________________________ 3% what? Nothing in any colloidal iron method involves mordanting, because no dye is involved (except as a counterstain, which might include a mordant as in the case of aluminium-nuclear fast red or brazalum). Mowry's technique is a modification of Hale's original, which wasn't much good. Pearse's Histochemistry reviews the various colloidal iron methods. It is the best reference, and has been translated into several languages. Is there any reason for using colloidal iron instead of an equivalent alcian blue method? Alcian blue techniques are more reproducible and have greater histochemical validity. Certified dye is available (tested in the Biological Stain Commission's lab, which is a non-profit outfit that serves both vendors and users of stains). The solid dye powder can deteriorate on the shelf, very unpredictably (I've had this happen 3 times). The deteriorated dye is insoluble; I have suspicions about why. Solutions of alcian blue at pH1 or pH3 keep working for a long time (a few years) even with repeated use. Alcian blue has its faults, but it's a better histochemical reagent than colloidal iron. There are several clever PAS variants developed by Scott et al in Britain and by Reid, Culling et al in Canada, from the late 1960s to the early 1990s. These methods have numerous steps but they stain macromolecular carbohydrates with well defined chemical specificity. These splendid methods are respectfully explained in textbooks but I suspect that they are seldom used in labs. There are also dozens of labeled lectins, which have high specificity and have been commercially available for 20+ years. There may be a lectin that binds specifically to the macromolecular carbohydrate that you are trying to stain with colloidal iron. Lectin histochemistry is EASY++ but you must do all the needed controls or your spectacularly specific pictures might be worthless artifacts. Lectins are the plant kingdoms's equivalent of antibodies. They recognize sugars and oligosaccharide sequences rather than the amino acid sequences (epitopes) that adhere to antibody molecules. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ASelf <@t> gmhsc.com Wed Oct 1 10:46:40 2003 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] HIPPA rules Message-ID: Hi all, Does anyone have any rules/regulations and or any policy/procedures that they can share with me as far as sending pathology slides and blocks out to other area hospitals for consult, research, etc? This has been on-going for us ever since HIPPA has come into affect. Thanks in advance for your help...... Amy Self Georgetown Memorial Hospital Georgetown, SC 843-520-7882 (fax) 843-527-7179 (phone) Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or 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 jincan <@t> itsa.ucsf.edu Wed Oct 1 11:10:54 2003 From: jincan <@t> itsa.ucsf.edu (jincan) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] FisherBiotech MicroProbe Staining Station Message-ID: <5.1.1.6.0.20031001085317.00b97df8@itsa.ucsf.edu> Dear Busy Colleagues, I would very much appreciate that you could share with us your experience with MicroProbe staining system from Fisher Scientific. We intend to get one for a research project processing only 10-20 slides per day. I would particularly like to know if staining results correlate closer to conventional manual staining or closer to Techmate system (also a capillary action system) as well as the overall performance of the MicroProbe. Thank you very much in advance! James Guo ImmunoVision Technologies, Co. 100 North Hill Drive, #32 Brisbane, CA 94005 Tel: 650-320-4622 Fax: 650-558-9621 From HornHV <@t> archildrens.org Wed Oct 1 11:20:00 2003 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] hardware and foreign bodies Message-ID: All foreign bodies are sent to the lab for documentation. We do not get the hardware from fractures, etc. We do return FB's to the patients if we are requested to do so. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: Satterfield, Marirose [mailto:marirose.satterfield@MercyMemorial.org] Sent: Wednesday, October 01, 2003 9:58 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] hardware and foreign bodies Our surgical doctors want to be able to give their patients their hardware and FB without it being grossed in the Histology Lab. I guess they say it would save the patient money by not having a pathology fee. I was wondering how other hospital handle their hardware and FB. Thanks in advance Mari Mercy Memorial Hospital _______________________________________________ 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. Arkansas Children's Hospital From Luis.Chiriboga <@t> med.nyu.edu Wed Oct 1 11:24:17 2003 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] HIPPA rules In-Reply-To: Message-ID: ya......if you do with out approval, you go to jail (sorry couldn't resist !!) Not sure about sending slides out for consult but for research purposes, there's "supposed" to be IRB approval between the institutions. However, the details of whether your sending slides vs. blocks etc....is still hazy. Your best bet is to check with your institution and the institution your collaborating with. Get ready for a lot of red tape.....good luck -------------------------------------- Luis Chiriboga Ph.D., HT (ASCP) QIHC New York University School Of Medicine NYU Cancer Institute and Bellevue Hospital Center Department Of Pathology 4W27 27th Street & First Avenue New York, N.Y. 10016 (212) 562-4667. -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Amy Self Sent: Wednesday, October 01, 2003 11:47 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] HIPPA rules Hi all, Does anyone have any rules/regulations and or any policy/procedures that they can share with me as far as sending pathology slides and blocks out to other area hospitals for consult, research, etc? This has been on-going for us ever since HIPPA has come into affect. Thanks in advance for your help...... Amy Self Georgetown Memorial Hospital Georgetown, SC 843-520-7882 (fax) 843-527-7179 (phone) Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or 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 pmarcum <@t> polysciences.com Wed Oct 1 11:07:05 2003 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Perfusion techniques in mice/rats In-Reply-To: <082C721AF78DB34983E8BA2CD085462105C695@mailbe07> Message-ID: <002501c38836$0ef49f70$4800a8c0@PMARCUM2K> If you do not have a pump available you can use an old fashion method. A glass bottle and stopper with two holes - one for the tubing and 18 gauge needle, sharp edge filed smooth (or size required) to be attached through and one for air to be admitted that is longer and is above the fluid level. Or a refillable IV bag from the pharmacy with tubing. The bag or bottle is hung at least 2 1/2 feet to 3 feet above the level of the animal to be perfused so make sure the tubing is long enough. The clamp on the tubing for fluid should be adjustable and set for a steady and slow drip to avoid vascular damage by rapid rate of flow. The methods for opening the chest and exposure of the heart are clear in all of the suggestions given. The descending aorta can be clamped to avoid perfusing the lower part of the body. (One thing about using a bottle - Remember to place rubber bands around the bottle over the stopper as they can come out and fixative goes everywhere.) Pam Marcum -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Davis, Gareth Sent: Wednesday, October 01, 2003 10:32 AM To: Histonet Subject: Re: [Histonet] Perfusion techniques in mice/rats Perfusion Protocol Preparation for Perfusion: Fixative =4% PFA Prepare fresh on day of use or the night before and keep at 4oC. 4% Paraformaldehyde 3.8% Sodium Borate Add 40g of paraformaldehyde to 800ml of water and heat to 60oC while stirring. Add 38g of sodium borate, paraformaldehyde will not dissolve until it is added. Cool off to 4oC. Adjust pH to 9.5 with glacial acetic acid. 4% PFA with 10% sucrose= In a 100ml cylinder add 10g of Sucrose, than add 4% PFA to 100ml Perfusion 1.. Anesthetize animal, open thoracic cavity, expose heart and visualize ascending aorta. Insert 18-gauge needle (on vacutainer collection tubing) through left ventricle into ascending aorta- if possible, hard to tell sometimes. Clamp cannula into place with would clip or small clamp and then snip right atrium. (I find it's easier - on mice- to hold needle in place, because clamp gets in the way). 2.. Slowly perfuse animal with about 40-50ml room temperature saline (it's recommended to use a pump here, but I use 50 ml syringe, and perfuse slowly) until fluid is clear. 3.. Change to cold 4% PFA and slowly perfuse animal. When the animal?s forelimbs are fully extended reduce perfusion speed, use about 50ml of fixative (for a 30g mouse). 4.. After perfusion is complete, decapitate the animal and quickly remove the brain using a caudal approach. Place brain in 4% PFA with 10% sucrose at 4oC for overnight. (The 4%PFA with sucrose will post-fix brain and cryoprotect tissue at the same time.) Some recommend to post-fix in 4% PFA-only, overnight, then cryoprotect with 30% sucrose for another 24 hours. Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/7e497724/attachment.htm From HornHV <@t> archildrens.org Wed Oct 1 11:43:12 2003 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] HIPPA rules Message-ID: ALL research HAS to have IRB approval. No exceptions, doesn't matter whether it's slides or blocks. But this would not apply to case reports for journals, etc. Consults are totally different. If it is your pathologist asking for a consult, no extra paperwork is needed. If it is the parents/patient then you need a signed release of patient information. I'm sure all hospitals handle this a bit different. Check with your HIPPA committee. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: Luis Chiriboga [mailto:Luis.Chiriboga@med.nyu.edu] Sent: Wednesday, October 01, 2003 11:24 AM To: Amy Self; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] HIPPA rules ya......if you do with out approval, you go to jail (sorry couldn't resist !!) Not sure about sending slides out for consult but for research purposes, there's "supposed" to be IRB approval between the institutions. However, the details of whether your sending slides vs. blocks etc....is still hazy. Your best bet is to check with your institution and the institution your collaborating with. Get ready for a lot of red tape.....good luck -------------------------------------- Luis Chiriboga Ph.D., HT (ASCP) QIHC New York University School Of Medicine NYU Cancer Institute and Bellevue Hospital Center Department Of Pathology 4W27 27th Street & First Avenue New York, N.Y. 10016 (212) 562-4667. -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Amy Self Sent: Wednesday, October 01, 2003 11:47 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] HIPPA rules Hi all, Does anyone have any rules/regulations and or any policy/procedures that they can share with me as far as sending pathology slides and blocks out to other area hospitals for consult, research, etc? This has been on-going for us ever since HIPPA has come into affect. Thanks in advance for your help...... Amy Self Georgetown Memorial Hospital Georgetown, SC 843-520-7882 (fax) 843-527-7179 (phone) Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or 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 _______________________________________________ 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. Arkansas Children's Hospital From pmarcum <@t> polysciences.com Wed Oct 1 11:07:05 2003 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Perfusion techniques in mice/rats In-Reply-To: <082C721AF78DB34983E8BA2CD085462105C695@mailbe07> Message-ID: <002501c38836$0ef49f70$4800a8c0@PMARCUM2K> If you do not have a pump available you can use an old fashion method. A glass bottle and stopper with two holes - one for the tubing and 18 gauge needle, sharp edge filed smooth (or size required) to be attached through and one for air to be admitted that is longer and is above the fluid level. Or a refillable IV bag from the pharmacy with tubing. The bag or bottle is hung at least 2 1/2 feet to 3 feet above the level of the animal to be perfused so make sure the tubing is long enough. The clamp on the tubing for fluid should be adjustable and set for a steady and slow drip to avoid vascular damage by rapid rate of flow. The methods for opening the chest and exposure of the heart are clear in all of the suggestions given. The descending aorta can be clamped to avoid perfusing the lower part of the body. (One thing about using a bottle - Remember to place rubber bands around the bottle over the stopper as they can come out and fixative goes everywhere.) Pam Marcum -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Davis, Gareth Sent: Wednesday, October 01, 2003 10:32 AM To: Histonet Subject: Re: [Histonet] Perfusion techniques in mice/rats Perfusion Protocol Preparation for Perfusion: Fixative =4% PFA Prepare fresh on day of use or the night before and keep at 4oC. 4% Paraformaldehyde 3.8% Sodium Borate Add 40g of paraformaldehyde to 800ml of water and heat to 60oC while stirring. Add 38g of sodium borate, paraformaldehyde will not dissolve until it is added. Cool off to 4oC. Adjust pH to 9.5 with glacial acetic acid. 4% PFA with 10% sucrose= In a 100ml cylinder add 10g of Sucrose, than add 4% PFA to 100ml Perfusion 1.. Anesthetize animal, open thoracic cavity, expose heart and visualize ascending aorta. Insert 18-gauge needle (on vacutainer collection tubing) through left ventricle into ascending aorta- if possible, hard to tell sometimes. Clamp cannula into place with would clip or small clamp and then snip right atrium. (I find it's easier - on mice- to hold needle in place, because clamp gets in the way). 2.. Slowly perfuse animal with about 40-50ml room temperature saline (it's recommended to use a pump here, but I use 50 ml syringe, and perfuse slowly) until fluid is clear. 3.. Change to cold 4% PFA and slowly perfuse animal. When the animal?s forelimbs are fully extended reduce perfusion speed, use about 50ml of fixative (for a 30g mouse). 4.. After perfusion is complete, decapitate the animal and quickly remove the brain using a caudal approach. Place brain in 4% PFA with 10% sucrose at 4oC for overnight. (The 4%PFA with sucrose will post-fix brain and cryoprotect tissue at the same time.) Some recommend to post-fix in 4% PFA-only, overnight, then cryoprotect with 30% sucrose for another 24 hours. Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/7e497724/attachment-0001.htm From Noel.Clark <@t> ORTHO.UAB.EDU Wed Oct 1 11:16:53 2003 From: Noel.Clark <@t> ORTHO.UAB.EDU (Noel D. Clark) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Perfusion techniques in mice/rats Message-ID: <85F6C7A1330E794DB8540AFD001CC77E0D6712@ROSCO> Sorry. Forgot to mention that we want to harvest bones after perfusion! Noel Noel Clark, M.A., HTL (ASCP) 1919 7th Ave South Orthopaedic Research Laboratory Center for Metabolic Bone Disease University of Alabama at Birmingham Birmingham, Alabama 35294 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/2ae5c7e9/attachment.htm From jkayal <@t> bellsouth.net Wed Oct 1 12:03:21 2003 From: jkayal <@t> bellsouth.net (John Kayal) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] (no subject) Message-ID: <3F7B08D9.18D8F99B@bellsouth.net> Histotechnologist position available in start-up solo dermatology practice Marietta, Ga. Experience cutting Mohs frozen sections preferred. Competitive compensation and benefits. Please forward cv to jkayal@bellsouth.net or fax to 404-365-6895. From Mauricio.Morais <@t> tufts.edu Wed Oct 1 12:11:22 2003 From: Mauricio.Morais <@t> tufts.edu (Mauricio S. Morais) Date: Fri Sep 16 15:21:58 2005 Subject: Thank you; Re: [Histonet] automated immunostainers References: <533E4A1C3061FC4EB388119D2FD7B5D402431809@nbfexch04.north-bristol.nhs> <3F79D88E.CA2938A1@tufts.edu> Message-ID: <3F7B0ABA.9A801B60@tufts.edu> Hello Histonetters. I want to thank all that reply my original message. They are all posted below for future reference if anyone else need it. Thanks. Mauricio S. Morais Original Message: Hello Histonetters. Can anyone give me an idea about the costs of the machines (system) plus monthly maintenance? I wish to know if worth to have one to process 6-10 slides/day. Thank you. Mauricio S. Morais Senior Research Technician Nutrition, Exercise, Physiology, and Sarcopenia Laboratory (NEPS) TUFTS University Jean Mayer USDA Human Nutrition Research Center on Aging 711 Washington Street, Rm 436 Boston, MA 02111 (617) 556-3226 Replies: 1- Mauricio, It may not seem that an instrument is necessary to process a small amount of slides, but you should ask yourself if having an uninterrupted block of time is worthwhile. In otherwords, instead of tending to your slides every few minutes for several hours, you could run an instrument and have that extended time to concentrate on something. Lab Vision makes relatively inexpensive immunostaining instruments (including a new 36-slide model) for $35K to $50K, but maybe that is not what you need. There are also some setups that make manual staining more efficient. Look at the ThermoShandon Sequenza immuno work station. http://www.thermo.com/eThermo/CDA/Products/Product_Detail/1,1075,10000056307 76-117-X-117-1000000008097,00.html It allows you to easily add reagents without handling the slides. It is faster than manual staining, but not expensive like an instrument. You might also look at the Biocare Medical Kinetic Slide Stainer. http://www.biocare.net/equipment.htm Again, it limits slide handling. Good luck! Tim Morken Product Development Lab Vision / NeoMarkers 47790 Westinghouse Dr Fremont, CA 94539 PH: 510-991-2840 www.labvision.com 2- Dear Mauricio, Greetings! I happen to see this request on the Histonet and though of responding to you directly. I represent Biocare Medical, which is relatively new company that offers Semi-Manual Kinetic Slide Stainer and other related Immunohistochemistry products. I am attaching a pdf literature on our IQ Kinetic stainer that could hold 24-36 slides on the slide rack (12 slides on each rack and comes in 2 or 3 racks). This slide holder eliminates the slide handling once loaded and has a Hot Bar that could be controlled individually between RT-99 Degree C (Programmable). For washes, the rack could be place in an inclined position on grooves thus eliminating slide handling. The kinetic effect provided by the shaker helps better reaction at a lower time. The reagents are to be added manually. Please read the prices on the 24 slides as $5795 in the literature. We also offer Polymer Detection and Double stain detection cocktails and kits. Visit our web site www.biocare.net for further details. If you any further questions or concerns, please do not hesitate to contact me. Best regards, Hari Kumar Technical Sales (Research Division) Biocare Medical Phone: (925) 952-3901 Fax: (925) 944-7888 hari@biocare.net Please visit us at www.biocare.net for the latest. 3- Hi Mauricio, I am the sales consultant for Cell Marque and have sent you a couple of attachments that gives details on the cost per slide with Cell Marque's products. We do not have an automated immunostainer but all of our products work for manual customers or automated customers. Please let me know if you have any questions or would like to receive a catalog from our company. Sincerely, Laura Hughes Cell Marque Corporation Technical Sales Consultant 4- DAKO autostainers run about $45,000 to buy and $5000/year in a maintenance contract. It holds 48 slides/run. You can use your own reagents. Ventana NeXes autostainers are about $20-30,000 to buy and $5000/year in a maintenance contract. It holds 20 slides/run. You have to use their reagents. Jan Shivers Scientist, IHC Lab U of MN Veterinary Diagnostic Lab "Mauricio S. Morais" wrote: > > Hello Histonetters. > > Can anyone give me an idea about the costs of the machines (system) plus > monthly maintenance? > I wish to know if worth to have one to process 6-10 slides/day. > > Thank you. > > Mauricio S. Morais > Senior Research Technician > Nutrition, Exercise, Physiology, and Sarcopenia Laboratory (NEPS) > TUFTS University > Jean Mayer USDA Human Nutrition Research Center on Aging > 711 Washington Street, Rm 436 > Boston, MA 02111 > (617) 556-3226 > -------------- next part -------------- A non-text attachment was scrubbed... Name: Mauricio.Morais.vcf Type: text/x-vcard Size: 1220 bytes Desc: Card for Mauricio S. Morais Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/30cf3436/Mauricio.Morais.vcf From pmarcum <@t> polysciences.com Wed Oct 1 11:07:05 2003 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Perfusion techniques in mice/rats In-Reply-To: <082C721AF78DB34983E8BA2CD085462105C695@mailbe07> Message-ID: <002501c38836$0ef49f70$4800a8c0@PMARCUM2K> If you do not have a pump available you can use an old fashion method. A glass bottle and stopper with two holes - one for the tubing and 18 gauge needle, sharp edge filed smooth (or size required) to be attached through and one for air to be admitted that is longer and is above the fluid level. Or a refillable IV bag from the pharmacy with tubing. The bag or bottle is hung at least 2 1/2 feet to 3 feet above the level of the animal to be perfused so make sure the tubing is long enough. The clamp on the tubing for fluid should be adjustable and set for a steady and slow drip to avoid vascular damage by rapid rate of flow. The methods for opening the chest and exposure of the heart are clear in all of the suggestions given. The descending aorta can be clamped to avoid perfusing the lower part of the body. (One thing about using a bottle - Remember to place rubber bands around the bottle over the stopper as they can come out and fixative goes everywhere.) Pam Marcum -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Davis, Gareth Sent: Wednesday, October 01, 2003 10:32 AM To: Histonet Subject: Re: [Histonet] Perfusion techniques in mice/rats Perfusion Protocol Preparation for Perfusion: Fixative =4% PFA Prepare fresh on day of use or the night before and keep at 4oC. 4% Paraformaldehyde 3.8% Sodium Borate Add 40g of paraformaldehyde to 800ml of water and heat to 60oC while stirring. Add 38g of sodium borate, paraformaldehyde will not dissolve until it is added. Cool off to 4oC. Adjust pH to 9.5 with glacial acetic acid. 4% PFA with 10% sucrose= In a 100ml cylinder add 10g of Sucrose, than add 4% PFA to 100ml Perfusion 1.. Anesthetize animal, open thoracic cavity, expose heart and visualize ascending aorta. Insert 18-gauge needle (on vacutainer collection tubing) through left ventricle into ascending aorta- if possible, hard to tell sometimes. Clamp cannula into place with would clip or small clamp and then snip right atrium. (I find it's easier - on mice- to hold needle in place, because clamp gets in the way). 2.. Slowly perfuse animal with about 40-50ml room temperature saline (it's recommended to use a pump here, but I use 50 ml syringe, and perfuse slowly) until fluid is clear. 3.. Change to cold 4% PFA and slowly perfuse animal. When the animal?s forelimbs are fully extended reduce perfusion speed, use about 50ml of fixative (for a 30g mouse). 4.. After perfusion is complete, decapitate the animal and quickly remove the brain using a caudal approach. Place brain in 4% PFA with 10% sucrose at 4oC for overnight. (The 4%PFA with sucrose will post-fix brain and cryoprotect tissue at the same time.) Some recommend to post-fix in 4% PFA-only, overnight, then cryoprotect with 30% sucrose for another 24 hours. Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/7e497724/attachment-0002.htm From jmacdona <@t> mtsac.edu Wed Oct 1 12:45:50 2003 From: jmacdona <@t> mtsac.edu (Jmacdona) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] sperm staining Message-ID: <20031001174550.CB22D3D4DF@zeus.mtsac.edu> An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/0c2978a1/attachment.htm From pmarcum <@t> polysciences.com Wed Oct 1 11:07:05 2003 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Perfusion techniques in mice/rats In-Reply-To: <082C721AF78DB34983E8BA2CD085462105C695@mailbe07> Message-ID: <002501c38836$0ef49f70$4800a8c0@PMARCUM2K> If you do not have a pump available you can use an old fashion method. A glass bottle and stopper with two holes - one for the tubing and 18 gauge needle, sharp edge filed smooth (or size required) to be attached through and one for air to be admitted that is longer and is above the fluid level. Or a refillable IV bag from the pharmacy with tubing. The bag or bottle is hung at least 2 1/2 feet to 3 feet above the level of the animal to be perfused so make sure the tubing is long enough. The clamp on the tubing for fluid should be adjustable and set for a steady and slow drip to avoid vascular damage by rapid rate of flow. The methods for opening the chest and exposure of the heart are clear in all of the suggestions given. The descending aorta can be clamped to avoid perfusing the lower part of the body. (One thing about using a bottle - Remember to place rubber bands around the bottle over the stopper as they can come out and fixative goes everywhere.) Pam Marcum -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Davis, Gareth Sent: Wednesday, October 01, 2003 10:32 AM To: Histonet Subject: Re: [Histonet] Perfusion techniques in mice/rats Perfusion Protocol Preparation for Perfusion: Fixative =4% PFA Prepare fresh on day of use or the night before and keep at 4oC. 4% Paraformaldehyde 3.8% Sodium Borate Add 40g of paraformaldehyde to 800ml of water and heat to 60oC while stirring. Add 38g of sodium borate, paraformaldehyde will not dissolve until it is added. Cool off to 4oC. Adjust pH to 9.5 with glacial acetic acid. 4% PFA with 10% sucrose= In a 100ml cylinder add 10g of Sucrose, than add 4% PFA to 100ml Perfusion 1.. Anesthetize animal, open thoracic cavity, expose heart and visualize ascending aorta. Insert 18-gauge needle (on vacutainer collection tubing) through left ventricle into ascending aorta- if possible, hard to tell sometimes. Clamp cannula into place with would clip or small clamp and then snip right atrium. (I find it's easier - on mice- to hold needle in place, because clamp gets in the way). 2.. Slowly perfuse animal with about 40-50ml room temperature saline (it's recommended to use a pump here, but I use 50 ml syringe, and perfuse slowly) until fluid is clear. 3.. Change to cold 4% PFA and slowly perfuse animal. When the animal?s forelimbs are fully extended reduce perfusion speed, use about 50ml of fixative (for a 30g mouse). 4.. After perfusion is complete, decapitate the animal and quickly remove the brain using a caudal approach. Place brain in 4% PFA with 10% sucrose at 4oC for overnight. (The 4%PFA with sucrose will post-fix brain and cryoprotect tissue at the same time.) Some recommend to post-fix in 4% PFA-only, overnight, then cryoprotect with 30% sucrose for another 24 hours. Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/7e497724/attachment-0003.htm From djohnso1 <@t> uci.edu Wed Oct 1 13:25:08 2003 From: djohnso1 <@t> uci.edu (Johnson, David) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Kaposi sarcoma Message-ID: Does anyone know when I can find an antibody for Kaposi sarcoma? The main vendors like Zymed, Dako, etc. don't list it. Thanks DB Johnson Univ. Calif. Irvine MC This e-mail/fax 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/fax and destroy all copies of the original message. From Ngilman <@t> nbhd.org Wed Oct 1 14:00:20 2003 From: Ngilman <@t> nbhd.org (Noreen Gilman) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] mast cells Message-ID: Can anyone recommend a stain for mast cells? The one we do just doesn't seem to be working as well as we'd like it to. thanks noreen Noreen Gilman, BS, HT (ASCP) QIHC Histopathology Supervisor Broward General Medical Center Ft. Lauderdale, FL 33316 954.355.4358 Phone 954.355.4139 Fax 954.387.0213 Pager -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/2076264c/attachment.htm From pmarcum <@t> polysciences.com Wed Oct 1 11:07:05 2003 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Perfusion techniques in mice/rats In-Reply-To: <082C721AF78DB34983E8BA2CD085462105C695@mailbe07> Message-ID: <002501c38836$0ef49f70$4800a8c0@PMARCUM2K> If you do not have a pump available you can use an old fashion method. A glass bottle and stopper with two holes - one for the tubing and 18 gauge needle, sharp edge filed smooth (or size required) to be attached through and one for air to be admitted that is longer and is above the fluid level. Or a refillable IV bag from the pharmacy with tubing. The bag or bottle is hung at least 2 1/2 feet to 3 feet above the level of the animal to be perfused so make sure the tubing is long enough. The clamp on the tubing for fluid should be adjustable and set for a steady and slow drip to avoid vascular damage by rapid rate of flow. The methods for opening the chest and exposure of the heart are clear in all of the suggestions given. The descending aorta can be clamped to avoid perfusing the lower part of the body. (One thing about using a bottle - Remember to place rubber bands around the bottle over the stopper as they can come out and fixative goes everywhere.) Pam Marcum -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Davis, Gareth Sent: Wednesday, October 01, 2003 10:32 AM To: Histonet Subject: Re: [Histonet] Perfusion techniques in mice/rats Perfusion Protocol Preparation for Perfusion: Fixative =4% PFA Prepare fresh on day of use or the night before and keep at 4oC. 4% Paraformaldehyde 3.8% Sodium Borate Add 40g of paraformaldehyde to 800ml of water and heat to 60oC while stirring. Add 38g of sodium borate, paraformaldehyde will not dissolve until it is added. Cool off to 4oC. Adjust pH to 9.5 with glacial acetic acid. 4% PFA with 10% sucrose= In a 100ml cylinder add 10g of Sucrose, than add 4% PFA to 100ml Perfusion 1.. Anesthetize animal, open thoracic cavity, expose heart and visualize ascending aorta. Insert 18-gauge needle (on vacutainer collection tubing) through left ventricle into ascending aorta- if possible, hard to tell sometimes. Clamp cannula into place with would clip or small clamp and then snip right atrium. (I find it's easier - on mice- to hold needle in place, because clamp gets in the way). 2.. Slowly perfuse animal with about 40-50ml room temperature saline (it's recommended to use a pump here, but I use 50 ml syringe, and perfuse slowly) until fluid is clear. 3.. Change to cold 4% PFA and slowly perfuse animal. When the animal?s forelimbs are fully extended reduce perfusion speed, use about 50ml of fixative (for a 30g mouse). 4.. After perfusion is complete, decapitate the animal and quickly remove the brain using a caudal approach. Place brain in 4% PFA with 10% sucrose at 4oC for overnight. (The 4%PFA with sucrose will post-fix brain and cryoprotect tissue at the same time.) Some recommend to post-fix in 4% PFA-only, overnight, then cryoprotect with 30% sucrose for another 24 hours. Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/7e497724/attachment-0004.htm From lchung <@t> ppmh.org Wed Oct 1 14:08:59 2003 From: lchung <@t> ppmh.org (Chung, Luong) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] sperm staining Message-ID: Try Dip-Qik or Wright's stain, or PAP Stain. Bruce Chung Anatomic Pathology Manager Ext. 26130 -----Original Message----- From: Jmacdona [mailto:jmacdona@mtsac.edu] Sent: Wednesday, October 01, 2003 1:46 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] sperm staining Does anyone have a good staining method for human spermatozoa? -- Jennifer MacDonald --------- Original Message -------- From: "Kathleen Spencer" To: "Noel D. Clark" Cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Perfusion techniques in mice/rats Date: 01/10/03 07:32 Attached is the cardiac perfusion protocol that we use regularly. Kathleen Spencer HT, ASCP UTHSC Memphis, TN On Wednesday, October 1, 2003, at 07:45 AM, Noel D. Clark wrote: > Was hoping to get input on the various perfusion techniques used by > other research labs for rodents. As always, any help is greatly > appreciated. See you inLouisville! > > > > Thanks in advance, > > Noel > > > > > > > > > > Noel Clark, M.A., HTL (ASCP) > > 1919 7th Ave South > > Orthopaedic Research Laboratory > > Center for Metabolic Bone Disease > > UniversityofAlabamaatBirmingham > > Birmingham,Alabama35294 > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet HIPAA Privacy Rule requires covered entities to safeguard certain Protected Health Information (PHI) related to a person's healthcare. Information being sent to you may include PHI, after appropriate consent, acknowledgement, or authorization from the patient or under circumstances that do not require patient authorization. You, the recipient, are obligated to maintain PHI in a safe and secure manner. You may not re-disclose this patient information without additional patient consent or as required by law. Unauthorized re-disclosure or failure to safeguard PHI could subject us, or you, to penalties described in federal (HIPAA) and state law. If you, the reader of this message, are not the intended recipient, or the employee or agent responsible to deliver it to the intended recipient, please notify us immediately and destroy the related message. Thanks for your help. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/f3c9cb9f/attachment.htm From gareth.davis <@t> Vanderbilt.Edu Wed Oct 1 14:19:56 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Perfusion techniques in mice/rats Message-ID: <082C721AF78DB34983E8BA2CD085462105C696@mailbe07> Thanks for information on perfusion without a pump. Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -----Original Message----- From: Pamela Marcum [mailto:pmarcum@polysciences.com] Sent: Wed 10/1/2003 11:07 AM To: Davis, Gareth; Histonet Cc: Subject: RE: [Histonet] Perfusion techniques in mice/rats If you do not have a pump available you can use an old fashion method. A glass bottle and stopper with two holes - one for the tubing and 18 gauge needle, sharp edge filed smooth (or size required) to be attached through and one for air to be admitted that is longer and is above the fluid level. Or a refillable IV bag from the pharmacy with tubing. The bag or bottle is hung at least 2 1/2 feet to 3 feet above the level of the animal to be perfused so make sure the tubing is long enough. The clamp on the tubing for fluid should be adjustable and set for a steady and slow drip to avoid vascular damage by rapid rate of flow. The methods for opening the chest and exposure of the heart are clear in all of the suggestions given. The descending aorta can be clamped to avoid perfusing the lower part of the body. (One thing about using a bottle - Remember to place rubber bands around the bottle over the stopper as they can come out and fixative goes everywhere.) Pam Marcum -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Davis, Gareth Sent: Wednesday, October 01, 2003 10:32 AM To: Histonet Subject: Re: [Histonet] Perfusion techniques in mice/rats Perfusion Protocol Preparation for Perfusion: Fixative =4% PFA Prepare fresh on day of use or the night before and keep at 4oC. 4% Paraformaldehyde 3.8% Sodium Borate Add 40g of paraformaldehyde to 800ml of water and heat to 60oC while stirring. Add 38g of sodium borate, paraformaldehyde will not dissolve until it is added. Cool off to 4oC. Adjust pH to 9.5 with glacial acetic acid. 4% PFA with 10% sucrose= In a 100ml cylinder add 10g of Sucrose, than add 4% PFA to 100ml Perfusion 1. Anesthetize animal, open thoracic cavity, expose heart and visualize ascending aorta. Insert 18-gauge needle (on vacutainer collection tubing) through left ventricle into ascending aorta- if possible, hard to tell sometimes. Clamp cannula into place with would clip or small clamp and then snip right atrium. (I find it's easier - on mice- to hold needle in place, because clamp gets in the way). 2. Slowly perfuse animal with about 40-50ml room temperature saline (it's recommended to use a pump here, but I use 50 ml syringe, and perfuse slowly) until fluid is clear. 3. Change to cold 4% PFA and slowly perfuse animal. When the animal?s forelimbs are fully extended reduce perfusion speed, use about 50ml of fixative (for a 30g mouse). 4. After perfusion is complete, decapitate the animal and quickly remove the brain using a caudal approach. Place brain in 4% PFA with 10% sucrose at 4oC for overnight. (The 4%PFA with sucrose will post-fix brain and cryoprotect tissue at the same time.) Some recommend to post-fix in 4% PFA-only, overnight, then cryoprotect with 30% sucrose for another 24 hours. Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/0a5d3264/attachment.htm From stancelb <@t> msn.com Wed Oct 1 14:58:30 2003 From: stancelb <@t> msn.com (Barbara Stancel) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Drying Slides Message-ID: We save all of our slides each week, Monday through Friday. They are put in numerical order in flats and left at room temperature over the weekend. On Monday, they are filed. No problems in over twenty years. Barbara Barbara Stancel, HTL(ASCP) USDA, FSIS, OPHS, Eastern Laboratory, Pathology >From: "Mary Reeves" >To: >Subject: [Histonet] Drying Slides >Date: Tue, 30 Sep 2003 14:18:01 -0400 > >Currently we incubate our slides after the case is final, before we file >them. What are other places doing? Air drying? How long do you air dry >them? If you incubate them, how long? and at what temperature? Thanks in >advance for the information. > >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 _________________________________________________________________ Instant message during games with MSN Messenger 6.0. Download it now FREE! http://msnmessenger-download.com From gcallis <@t> montana.edu Wed Oct 1 15:18:17 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Microprobe versus Techmate, other manual devices Message-ID: <3.0.6.32.20031001141817.00ba0328@gemini.msu.montana.edu> James, We have used the Microprobe system (Fisher) which is merely a much downsized manual version of Techmate and definitely past tense on usage. It works well for immunostaining although we prefer to monitor our chromogen daily with microscope which means having to pull the positive control out and apart for viewing. Personally I think it works best with paraffin sections but fragile frozen sections need careful handling to pull slides apart when IHC is finished. Look into Chemicons new capillary gap stainer. This is a larger version of 20 slide capacity Microprobe holders - Chemicons holds 40 slides! If I had to do it over again and had Chemicon stainer available, we would buy it for the 40 slide capacity - it doesn't mean you HAVE to fill all the slots (we sometimes did 6 slides with Microprobe, but down the line, you will find you stain more and more sections. There are clever inexpensive ways to wick away buffers, etc - one does not need to buy fancy/pricey blotting pads nor buffer isolons (use larger dish reservoirs). Capillary gap is very cost effective in terms of reagent and spatial use. We never used the heater. One must become accustomed to capillary gap method - be sure to check Histonet Archives - some people loved it, others disliked it. As far as manual staining goes, we prefer to use Scytek SlideMaster flat, manual system, and set up 20 slides/holder or all 5 holders for 100 slides in a morning. This conventional manual stainer is so popular here lab that people are forever scrambling to use it - we love them! We find Slidemaster far less work than capillary gap, no loading more expensive Probe On slides face to face, no trapped bubbles, easier control of chromogen development, no filling isolons, etc - but the gap system works IF you want to take the time to use it. Good luck Dear Busy Colleagues, I would very much appreciate that you could share with us your experience with MicroProbe staining system from Fisher Scientific. We intend to get one for a research project processing only 10-20 slides per day. I would particularly like to know if staining results correlate closer to conventional manual staining or closer to Techmate system (also a capillary action system) as well as the overall performance of the MicroProbe. Thank you very much in advance! James Guo ImmunoVision Technologies, Co. 100 North Hill Drive, #32 Brisbane, CA 94005 Tel: 650-320-4622 Fax: 650-558-9621 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 Wed Oct 1 15:26:24 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Mouse perfusion Message-ID: <3.0.6.32.20031001142624.00ba0328@gemini.msu.montana.edu> Gareth Davis put out this a wonderful description of mouse perfusion on Histonet recently. It is much appreciated and worth its weight in gold!! Perfusion Protocol Preparation for Perfusion: Fixative =4% PFA Prepare fresh on day of use or the night before and keep at 4oC. 4% Paraformaldehyde 3.8% Sodium Borate Add 40g of paraformaldehyde to 800ml of water and heat to 60oC while stirring. Add 38g of sodium borate, paraformaldehyde will not dissolve until it is added. Cool off to 4oC. Adjust pH to 9.5 with glacial acetic acid. 4% PFA with 10% sucrose= In a 100ml cylinder add 10g of Sucrose, than add 4% PFA to 100ml Perfusion * Anesthetize animal, open thoracic cavity, expose heart and visualize ascending aorta. Insert 18-gauge needle (on vacutainer collection tubing) through left ventricle into ascending aorta- if possible, hard to tell sometimes. Clamp cannula into place with would clip or small clamp and then snip right atrium. (I find it's easier - on mice- to hold needle in place, because clamp gets in the way). * Slowly perfuse animal with about 40-50ml room temperature saline (it's recommended to use a pump here, but I use 50 ml syringe, and perfuse slowly) until fluid is clear. * Change to cold 4% PFA and slowly perfuse animal. When the animal???s forelimbs are fully extended reduce perfusion speed, use about 50ml of fixative (for a 30g mouse). * After perfusion is complete, decapitate the animal and quickly remove the brain using a caudal approach. Place brain in 4% PFA with 10% sucrose at 4oC for overnight. (The 4%PFA with sucrose will post-fix brain and cryoprotect tissue at the same time.) Some recommend to post-fix in 4% PFA-only, overnight, then cryoprotect with 30% sucrose for another 24 hours. Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 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 louri_c <@t> hotmail.com Wed Oct 1 15:28:00 2003 From: louri_c <@t> hotmail.com (Louri Caldwell) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Thin Preps Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/d6d7c3ea/attachment.htm From LuckG <@t> empirehealth.org Wed Oct 1 15:31:52 2003 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] mast cells Message-ID: Noreen, We use a Toluidine Blue stain which is very popular with our pathologists, although seldom used. Reference: Churukian, C.J., B.A. & Schenk, E.A., M.D.: "A Toluidine Blue Method of Demonstrating Mast Cells", Journal of Histotechnology, 4:2,pp 85-86, June 1981. Greg Luck Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 509.473.7077 luckg@empirehealth.org -----Original Message----- From: Noreen Gilman [mailto:Ngilman@nbhd.org] Sent: Wednesday, October 01, 2003 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] mast cells Can anyone recommend a stain for mast cells? The one we do just doesn't seem to be working as well as we'd like it to. thanks noreen Noreen Gilman, BS, HT (ASCP) QIHC Histopathology Supervisor Broward General Medical Center Ft. Lauderdale, FL 33316 954.355.4358 Phone 954.355.4139 Fax 954.387.0213 Pager -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/daaa19ad/attachment.htm From nick.kirk3 <@t> btopenworld.com Wed Oct 1 15:53:40 2003 From: nick.kirk3 <@t> btopenworld.com (Nick Kirk) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] mast cells In-Reply-To: Message-ID: We use a simple 1% aqueous Toluidine blue technique. Stain in the solution for about 10-15 minutes, blot dry and mount. Mast cell granules purple, rest of cells/tissue blue. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Noreen Gilman Sent: 01 October 2003 20:00 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] mast cells Can anyone recommend a stain for mast cells? The one we do just doesn't seem to be working as well as we'd like it to. thanks noreen Noreen Gilman, BS, HT (ASCP) QIHC Histopathology Supervisor Broward General Medical Center Ft. Lauderdale, FL 33316 954.355.4358 Phone 954.355.4139 Fax 954.387.0213 Pager -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/33094f54/attachment.htm From mrakas <@t> email.smith.edu Wed Oct 1 16:06:28 2003 From: mrakas <@t> email.smith.edu (Margaret Rakas) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Help ID'ing Really Old Histology Stains Message-ID: Hello, I am the Chemical Hygiene Officer and have been trying to obtain MSDS's for a number of old---and I mean 30 years or more--histological stains in order to properly characterize them for disposal. I've been to stainsfile, hazard.com, google, you name it. I don't know if the names have seriously changed, if these haven't been in use for the past 10 years (or more), etc. Any information you can provide would be most appreciated! The labels for these materials do not carry CAS #'s... The stains in question are: Aniline Red Acridine Red Chicago Blue RW Bordeaux Red Cyanine Renol Black R Extra (another name on its label is "Chlorazol Black R" Methylene Blue Eosin Many thanks! Margaret Margaret A. Rakas, Ph.D. Manager, Inventory & Regulatory Affairs Clark Science Center Smith College Northampton, MA. 01063 p: 413-585-3877 f: 413-585-3786 From lalsing <@t> cvdls.ucdavis.edu Wed Oct 1 16:15:54 2003 From: lalsing <@t> cvdls.ucdavis.edu (Elena Alsing) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Bird feather Message-ID: Greetings to all! How do you make bird feather stay on the slide? We received feather shafts in formalin. I decalcified them in equal amount of sodium citrate and formic acid and then process them. They cut nicely but they keep on lifting off the slide. I put them on plus slides and air dried them over the weekend - didn't work. I put them on probe-on slides, let them dry dry overnight at room temp and hand-stained them - didn't work. I don't know what to do... Please HELP! Thanks. Lalen Alsing UC Davis-CAHFS, Fresno From bills <@t> icpmr.wsahs.nsw.gov.au Wed Oct 1 16:31:29 2003 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Superfrost Plus Slides Message-ID: <002a01c38863$610e1ac0$8267080a@w2kbills> Dear All, Has anyone had a problem with Superfrost Plus slides on automated Immunohistochemistry Staining machines? If so what have the problems been? Also if the problem was resolved and how it was resolved. We obtain our slides through a third party agency in Australia. We are at present using Menzel-Glaser (German) brand. I am led to believe they are the largest slide and coverglass manufacturer in the world. 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 garygill <@t> dcla.com Wed Oct 1 16:37:06 2003 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Help ID'ing Really Old Histology Stains Message-ID: * aniline red yellow and purple, no red * acridine red [3B {CI 45000}] pyronin (basic dye) * Chicago blue RW (CI 24280) aka benzo blue RW, Niagra blue RW, durazol blue 5G * Bordeaux red (CI 16180) aka fast red B, BN or P; cerasin R * Cyanine (CI [Ed. 1] 806) aka quinoline blue (basic quinoline dye) * Renol black R extra (chlorazol black R) chlorazol black E (CI 30235) * methylene blue eosin ? All are benzene-ring based. Source: Lillie RD. H.J. Conn's Biological Stains, Williams & Wilkins, Baltimore, 1977, 9th ed. (10th edition has just been published). Dispose of these the same as you would comparable dyes for which you have information. If you're disposing of small quantities such as usually found in labs, it's not worth your time to research further. Gary Gill -----Original Message----- From: Margaret Rakas [mailto:mrakas@email.smith.edu] Sent: Wednesday, October 01, 2003 4:06 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Help ID'ing Really Old Histology Stains Hello, I am the Chemical Hygiene Officer and have been trying to obtain MSDS's for a number of old---and I mean 30 years or more--histological stains in order to properly characterize them for disposal. I've been to stainsfile, hazard.com, google, you name it. I don't know if the names have seriously changed, if these haven't been in use for the past 10 years (or more), etc. Any information you can provide would be most appreciated! The labels for these materials do not carry CAS #'s... The stains in question are: Aniline Red Acridine Red Chicago Blue RW Bordeaux Red Cyanine Renol Black R Extra (another name on its label is "Chlorazol Black R" Methylene Blue Eosin Many thanks! Margaret Margaret A. Rakas, Ph.D. Manager, Inventory & Regulatory Affairs Clark Science Center Smith College Northampton, MA. 01063 p: 413-585-3877 f: 413-585-3786 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RFail <@t> Charleston.net Wed Oct 1 17:28:59 2003 From: RFail <@t> Charleston.net (Rena Fail) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] mast cells In-Reply-To: Message-ID: <01cd01c3886b$697eed30$b611a6a5@rena> 2 A Giemsa or a Von Leder, the protocol for the Giemsa can be found in Histologic Rena Fail -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Noreen Gilman Sent: Wednesday, October 01, 2003 3:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] mast cells Can anyone recommend a stain for mast cells? The one we do just doesn't seem to be working as well as we'd like it to. thanks noreen Noreen Gilman, BS, HT (ASCP) QIHC Histopathology Supervisor Broward General Medical Center Ft. Lauderdale, FL 33316 954.355.4358 Phone 954.355.4139 Fax 954.387.0213 Pager -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/f1bbf1ad/attachment.htm From ploykasek <@t> phenopath.com Wed Oct 1 17:52:05 2003 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Kaposi sarcoma In-Reply-To: Message-ID: While I do not know of an antibody specific for KS, there is an antibody for HHV8. Almost all KS is positive with this anitbody. The antibody is clone LN-1 from Advanced Biotech., catalog #12-210-001. A positive signal yields a speckled nuclear pattern. I would suggest that you do your own in-house validation with this antnibody and KS. If you need any other info, please contact me. Patti Loykasek PhenoPath Laboratories Seattle, WA > Does anyone know when I can find an antibody for Kaposi sarcoma? The main > vendors like Zymed, Dako, etc. don't list it. Thanks > > DB Johnson > Univ. Calif. Irvine MC > > > This e-mail/fax 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/fax and destroy all copies of the original message. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From histo20 <@t> hotmail.com Wed Oct 1 18:39:15 2003 From: histo20 <@t> hotmail.com (Paula Wilder) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] BRST2 Message-ID: Can anyone tell me how they handle this antibody? We are getting quite a mixed response from neighboring institutions. Thanks so much! Paula Wilder St. Joseph Medical Center 7601 Osler Drive Towson, MD. _________________________________________________________________ Get McAfee virus scanning and cleaning of incoming attachments. Get Hotmail Extra Storage! http://join.msn.com/?PAGE=features/es From bamur <@t> alaska.net Wed Oct 1 06:40:59 2003 From: bamur <@t> alaska.net (Barbara Murray) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Retic Stain Message-ID: <000a01c38810$e5c43040$368b70d1@d3b1l9> Greetings, Our Retic stain need to be improved. May I have some techniques on what everyone else is doing? We are leaving it in the Silver Solution for 1 minute and the Gold Chloride for 5 mins. Everything else for 1 minute and washings for 2 minutes. Thanks in advance for all your suggestions. Have a great day! Barbara. A. Murray, HT. (ASCP) The Alaska Native Medical Center Pathology Dept. Anchorage, Alaska -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/f5bf2a14/attachment.htm From histo20 <@t> hotmail.com Wed Oct 1 18:42:11 2003 From: histo20 <@t> hotmail.com (Paula Wilder) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] ergonomic chairs Message-ID: I posted this earlier, but I think I deleted any messages which may have been sent. Does anyone have a suggestion concerning ergonomic chairs - which company did you like best, which style of chair did you like best? Any information will be greatly appreciated. Thanks so much! Paula Wilder 7601 Osler Drive Towson, MD. 21204 _________________________________________________________________ Instant message with integrated webcam using MSN Messenger 6.0. Try it now FREE! http://msnmessenger-download.com From ihctech2000 <@t> yahoo.com Wed Oct 1 18:44:47 2003 From: ihctech2000 <@t> yahoo.com (Sun Zhon) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] morphology on frozen tissues Message-ID: <20031001234447.631.qmail@web80706.mail.yahoo.com> Dear All, These were the steps I processed my frozen tissues, after sectioning, air dry tissues for 30 mins fix in acetone for 10 mins at RT air dry for 30 mins store in -80 freezer The next morning before I did my IHC, I took out the fixed frozen tissues from the -80 freezer and put them directly in PBS without letting them come to room temperature. My tissues looked like spider webs, especially around the edge. However, in another experiment I used exactly the same tissues and the tissues looked beautiful. The only difference was that while quenching the peroxidase, 0.03% H2O2 in PBS was used on the 1st exp (spider web morphology); 0.03% in 10% horse serum in 3% BSA was used on the 2nd exp (beautiful morphology). Do you think the H2O2 will make such a big difference on the morphology or there are some other reasons for the spider web effect? How do you fix and store your frozen tissues? How do you treat the tissues before you do your IHC exps? Thank you very much for your help. Sun Zhon --------------------------------- Do you Yahoo!? The New Yahoo! Shopping - with improved product search -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/96088f1b/attachment.htm From kb2drkprk <@t> yahoo.com Wed Oct 1 18:59:30 2003 From: kb2drkprk <@t> yahoo.com (Kelly Booher) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Retic Stain In-Reply-To: <000a01c38810$e5c43040$368b70d1@d3b1l9> Message-ID: <20031001235930.71467.qmail@web20913.mail.yahoo.com> Barbara, Is your Retic staining too light? We leave our slides in ammoniacal silver longer, and tone in gold chloride for a shorter period of time. Our procedure is as follows: 0.5% Potassium Permanganate - 1 minute Wash in tap water - 2 minutes 2% Potassium Metabisulfite - 1 minute Wash in tap water - 2 minutes 2% Ferric Ammonium Sulfate - 3 minutes Wash in tap water - 2 minutes Rinse in deionized water Ammonium Silver Nitrate - 3 minutes Rinse in deionized water - 10 seconds 20% Formalin - 3 minutes Wash in tap water - 3 minutes 0.2% Gold Chloride - 30 seconds to 1 minute Rinse in deionized water 2% Potassium Metabisulfite - 1 minute 2% Sodium Thiosulfate - 1 minute Wash in tap water - 2 minutes Dehydrate, clear, and mount Kelly Booher, BS, HTL (ASCP) Anatomic Pathology Swedish Medical Center, Providence Campus Seattle, WA 98122 --- Barbara Murray wrote: > Greetings, > Our Retic stain need to be improved. May I have > some techniques on what everyone else is doing? We > are leaving it in the Silver Solution for 1 minute > and the Gold Chloride for 5 mins. Everything else > for 1 minute and washings for 2 minutes. > Thanks in advance for all your suggestions. > Have a great day! > > Barbara. A. Murray, HT. (ASCP) > The Alaska Native Medical Center > Pathology Dept. > Anchorage, Alaska > > __________________________________ Do you Yahoo!? The New Yahoo! Shopping - with improved product search http://shopping.yahoo.com From GREYTRUNK <@t> aol.com Wed Oct 1 20:11:22 2003 From: GREYTRUNK <@t> aol.com (GREYTRUNK@aol.com) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] mast cells Message-ID: <151.24c0eae5.2cacd53a@aol.com> You can also use toludine blue. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031001/e9c17329/attachment.htm From histo <@t> skm.org.pk Wed Oct 1 22:04:04 2003 From: histo <@t> skm.org.pk (Histology) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] RE: Purchasing machines Grossing Work Station,Microtom etc. Message-ID: Our lab is in the process of purchasing our automated stainer for routine H&E's,Grossing Work Station,Microtom,Cryotom,Embedding center and processor (300 cassettes). I would really appreciate comments from anyone who really likes, or dislikes the machines that they are using. Our plan is to get an automatic coverslipper as well. Thanks in advance!!! Muhammad Tahseen Histology Supervisor Deptt. Pathology Shaukat Khanum Cancer Hospital And Research Center Lahore. Pakistan Ph. +92 42 5180725-36 Ext 2369, Fax. +92 42 5180723 e-mail. Histology@skm.org.pk From SJones <@t> cvm.tamu.edu Wed Oct 1 23:20:54 2003 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Bird feather Message-ID: Have you tried celloidin coating the sections after deparaffinization? Are they lifting in the oven before deparaffinization? I have the same trouble with horse hoof. I use the ultrastick goldseal slides, but still have trouble from time to time. You could try plastic embedding as a last resort. Sarah 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 >>> "Elena Alsing" 10/01/03 04:15PM >>> Greetings to all! How do you make bird feather stay on the slide? We received feather shafts in formalin. I decalcified them in equal amount of sodium citrate and formic acid and then process them. They cut nicely but they keep on lifting off the slide. I put them on plus slides and air dried them over the weekend - didn't work. I put them on probe-on slides, let them dry dry overnight at room temp and hand-stained them - didn't work. I don't know what to do... Please HELP! Thanks. Lalen Alsing UC Davis-CAHFS, Fresno _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SJones <@t> cvm.tamu.edu Wed Oct 1 23:34:05 2003 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] ergonomic chairs Message-ID: Hi Paula, We bought some from Champion Seating Company, phone 281-403-1777, http://www.championseating.com The chairs we bought were the MVP high-back chair with arms. They have been great back savers. 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 >>> "Paula Wilder" 10/01/03 06:42PM >>> I posted this earlier, but I think I deleted any messages which may have been sent. Does anyone have a suggestion concerning ergonomic chairs - which company did you like best, which style of chair did you like best? Any information will be greatly appreciated. Thanks so much! Paula Wilder 7601 Osler Drive Towson, MD. 21204 _________________________________________________________________ Instant message with integrated webcam using MSN Messenger 6.0. Try it now FREE! http://msnmessenger-download.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DDDeltour <@t> sig.med.navy.mil Wed Oct 1 23:57:11 2003 From: DDDeltour <@t> sig.med.navy.mil (Deltour, Douglas D.(HM2)) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] SAKURA TISSUE -TEK SCA Coverslipper Message-ID: I have worked with several of them. They are very easy machines to operate and the results are great. Very low maintenance. The machine also uses an air pump that pumps the medium to the slide. This pump will cost you much money if you get it through Sakura. It can be replaced with an air pump that is found at a pet store in the fish department for much less. We always keep one on hand just in case. The coverslipping tape is a little expensive when you use the Sakura brand. I know that https://www1.fishersci.com/ has the same type of coverslip tape for a better price. Here is the catalog number 15-182-52 Vendor No.:14-K-KP. Also Sakura offers the bottles of cover slipping medium but I have always used xylene with the same results. Let me know if you have any more questions. HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy LPO Histology Histology Technician DSN 624-4669 FAX 624-4680 COMM (From US) 011-39-095-56-4669 -----Original Message----- From: Histology [mailto:histo@skm.org.pk] Sent: Wednesday, October 01, 2003 4:38 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] SAKURA TISSUE -TEK SCA Coverslipper > Dear Histonetters, > I am considering purchasing the SAKURA TISSUE -TEK SCA Coverslipper. > I'd appreciate the hearing about your experiences. > Thanks in advanced, > Muhammad Tahseen Histology Supervisor Deptt. Pathology Shaukat Khanum Cancer Hospital And Research Center Lahore. Pakistan Ph. +92 42 5180725-36 Ext 2369, Fax. +92 42 5180723 e-mail. Histology@skm.org.pk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From docmichel <@t> netbulmail.com Thu Oct 2 01:28:51 2003 From: docmichel <@t> netbulmail.com (izzet oguz) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] need help about frozen bladder sections Message-ID: <3344.193.255.128.130.1065076131.webmail@mail1.netbulmail.com> we took bladder sections from rats about four months ago..first we froze the tissue in liquid nitrogen. Then left them at -20 C deep freeze for 4 months..after that we could not cut the tissues because they became so fragile..Any suggestions about this problem will be greatly appreciated.. Thanks.. ?zzet From kemlo <@t> tiscali.co.uk Thu Oct 2 04:52:17 2003 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] Thin Preps In-Reply-To: Message-ID: <000001c388ca$de2fed00$45c4e150@KEMLOS> ThinPrep equipment (Cytyc) is needed to prepare ThinPreps and there is no practical way of doing it manually, to my knowledge. There is another system (SurePath) that can prepare liquid based cytology preparations and it appears to be less expensive and some may say, more efficient (some would say, I'm not saying anything!!!!!!!!) However, look at both alternatives, the end result, some would say are similar (I'm not saying anything), but one thing I WILL say, is that either are better than conventional. I screen SurePath, ThinPrep and conventional every day. I prefer LBC over conventional but I'm not saying anything else in an open Forum! Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 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-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Louri Caldwell Sent: 01 October 2003 21:28 To: histonet@pathology.swmed.edu Subject: [Histonet] Thin Preps Hi everyone, Since Histology & Cytology sometimes cross, would any of you know if the Thin Prep equipment is required to prepare the slides for interpretation, or if there is a manual procedure for doing so? Any assistance you may provide is greatly appreciated. Thank you in advance for your help. Louri _____ Instant message during games with MSN Messenger 6.0. Download it now FREE! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031002/1fdb233f/attachment.htm From Wimer.Helen <@t> NMNH.SI.EDU Thu Oct 2 05:25:06 2003 From: Wimer.Helen <@t> NMNH.SI.EDU (Helen Wimer) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] sperm staining Message-ID: Have you tried the "Berg's stain"? Helen F. Wimer Wimer.Helen@NMNH.SI.EDU Smithsonian Institution MSC NW Washington , DC 20560 (301) 238-3786 From Luis.Chiriboga <@t> med.nyu.edu Thu Oct 2 06:10:48 2003 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] sperm staining In-Reply-To: <20031001174550.CB22D3D4DF@zeus.mtsac.edu> Message-ID: I believe it's called the Christmas stain, used by the forensic community -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Jmacdona Sent: Wednesday, October 01, 2003 1:46 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] sperm staining Does anyone have a good staining method for human spermatozoa? -- Jennifer MacDonald --------- Original Message -------- From: "Kathleen Spencer" To: "Noel D. Clark" Cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Perfusion techniques in mice/rats Date: 01/10/03 07:32 Attached is the cardiac perfusion protocol that we use regularly. Kathleen Spencer HT, ASCP UTHSC Memphis, TN On Wednesday, October 1, 2003, at 07:45 AM, Noel D. Clark wrote: > Was hoping to get input on the various perfusion techniques used by > other research labs for rodents. As always, any help is greatly > appreciated. See you inLouisville! > > > > Thanks in advance, > > Noel > > > > > > > > > > Noel Clark, M.A., HTL (ASCP) > > 1919 7th Ave South > > Orthopaedic Research Laboratory > > Center for Metabolic Bone Disease > > UniversityofAlabamaatBirmingham > > Birmingham,Alabama35294 > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031002/2486b586/attachment.htm From ree3 <@t> leicester.ac.uk Thu Oct 2 06:31:05 2003 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:21:58 2005 Subject: [Histonet] A6 antigen/mouse oval cells Message-ID: Wanted a supplier of an antibody to the mouse A6 antigen found in hepatic oval cells. Thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER....U.K....... -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031002/25c73de8/attachment.htm From Nancy.Walker <@t> sanofi-synthelabo.com Thu Oct 2 07:33:06 2003 From: Nancy.Walker <@t> sanofi-synthelabo.com (Nancy.Walker@sanofi-synthelabo.com) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] embryo perfusion Message-ID: Hello, Need help on PFA perfusion techniques on rat and mice embryos (E18). I've tried perfusing (intracardiaque) the mothers (10 minutes with PBS/heparine then 30 minutes with PFA (pump speed of 2ml /min for mice and 25ml/min for rats), spread the babies out on the table...one sees that the saline solution does clear the blood, and the PFA does harden them up. I then make a sagital slice through each embryo and post fix for 24h followed by paraffin embedding. I then make 7? sagittal slices. But in the stained sections there are still an abundance of red blood cells that are a big source of background for IHC, and the tissue seems underfixed because there is shrinkage (alcohol fixation during embedding). I've heard that one can perfuse individual embryos via the umbelical cord. Does anyone have a description of this with a schema or explanation on how to find the right artery, pump speed and PFA volume. thanks for your time, Nancy Walker Molecular Biology Scientist Sanofi-Synthelbo Research B.P. 37 Lab?ge Innopole 31676 LABEGE CEDEX FRANCE nancy.walker@sanofi-synthelabo.com tel : (33)561004179? fax :(33)561004001 From van <@t> uhnres.utoronto.ca Thu Oct 2 08:04:12 2003 From: van <@t> uhnres.utoronto.ca (Rita van Bendegem) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Superfrost Plus Slides References: <002a01c38863$610e1ac0$8267080a@w2kbills> Message-ID: <3F7C224B.C08E20D3@uhnres.utoronto.ca> Dear All, I have had some problems with Superfrosted slides but in our research lab we process our IHC slides manual. We react both frozens and paraffin tissues. Our routine chromogens that we use are DAB, VIP, Alexa 488, and Alexa 594, depending on the tissue and the application. We have been using Fisherbrand Superfrosted Plus slides for years with no known problems. In February of this year I discovered that NF200 antibody from Sigma was not working at all in frozen or paraffin tissue, but my positive control tissue was O.K. (since it was cut on to older Superfrosted slides) I pulled out some old paraffin slides dating back to Jan. 2002 and Sept. 2002 then ran the NF200 immuno. The Jan. 2002 slides were positive, Sept. 2002 were weakly positive and I also ran another Feb. 2003 slide which was negative. This made me call my Fisher rep. about the problems and he in return called the manufacture of the Superfrosted Plus slides, which from my understanding is ERIE in North America. The Fisher rep. told me that he never got a response back from the manufacture as of a few months ago. In the meantime, I called a few techs and made some inquires. I was told to try some slides from Surgipath that are called X-tra Adhesive they are European slides with USA, Canada, France and UK address written on the side of box. I have used them and have had no problems with my NF200 controls since. I would be interested to hear if anyone else has had similar problems . Bill Sinai wrote: > Dear All, > > Has anyone had a problem with Superfrost Plus slides on automated > Immunohistochemistry Staining machines? If so what have the problems been? > Also if the problem was resolved and how it was resolved. > > We obtain our slides through a third party agency in Australia. We are at > present using Menzel-Glaser (German) brand. I am led to believe they are > the largest slide and coverglass manufacturer in the world. > > 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. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Rita van Bendegem, MLT Laboratory Manager, Canadian Paraplegic Association, Spinal Cord Injury Research Centre, Mc Laughlin Pavilion Lab 12-405, 399 Bathurst St., Toronto Western Hospital, University Health Network, Toronto, Ontario M5T 2S8 Telephone 416-603-5800 ext. 2792 Fax 416-603-5745 From cwscouten <@t> myneurolab.com Thu Oct 2 08:01:41 2003 From: cwscouten <@t> myneurolab.com (Charles W. Scouten, Ph.D.) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] embryo perfusion Message-ID: Even neonates, probably embryo's, have a blood pressure comparable to other mammals, up above 100 mm Hg. systolic. 2 ml/s per minute will generate how much pressure? You need to be comparable to physiological pressure or above, or you will not get the red blood cells out. Too far above will damage tissue. I would guess from the rate and results that you are way below physiological pressure, maybe 20 mm Hg. The capillaries won't open and release their blood. Then fixative will not flow through the occupied capillaries. Try controlling your pressure of perfusion, not your flow rate, unless you know what flow rate will generate what pressure. Use at least 150 mm Hg. To avoid shrinkage, use the Perfusion One system, see the link: http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=471001&catdesc=Histology+Equipment&CatThreeID=674&CatOneID=4&subcatdesc=Sacrifice+Equipment&idsubcategory=21 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 www.myneurolab.com -----Original Message----- From: Nancy.Walker@sanofi-synthelabo.com [mailto:Nancy.Walker@sanofi-synthelabo.com] Sent: Thursday, October 02, 2003 7:33 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] embryo perfusion Hello, Need help on PFA perfusion techniques on rat and mice embryos (E18). I've tried perfusing (intracardiaque) the mothers (10 minutes with PBS/heparine then 30 minutes with PFA (pump speed of 2ml /min for mice and 25ml/min for rats), spread the babies out on the table...one sees that the saline solution does clear the blood, and the PFA does harden them up. I then make a sagital slice through each embryo and post fix for 24h followed by paraffin embedding. I then make 7? sagittal slices. But in the stained sections there are still an abundance of red blood cells that are a big source of background for IHC, and the tissue seems underfixed because there is shrinkage (alcohol fixation during embedding). I've heard that one can perfuse individual embryos via the umbelical cord. Does anyone have a description of this with a schema or explanation on how to find the right artery, pump speed and PFA volume. thanks for your time, Nancy Walker Molecular Biology Scientist Sanofi-Synthelbo Research B.P. 37 Lab?ge Innopole 31676 LABEGE CEDEX FRANCE nancy.walker@sanofi-synthelabo.com tel : (33)561004179? fax :(33)561004001 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gauchv <@t> mail.amc.edu Thu Oct 2 08:37:31 2003 From: gauchv <@t> mail.amc.edu (Vicki Gauch) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] RE: Purchasing machines Grossing Work Station,Microtom etc. Message-ID: We have the Shandon Gross Lab Senior as the grossing station in our Frozen Section/Grossing Room and love it. The adjustable height feature has been very useful since we have residents and technical staff ranging from 5'2" to 6'0"...so everyone can work at a height that is comfortable for them . The hands free sensor sytem for turning on the water in the sink, the disposal unit and surface rinse are also wonderful. The ventilation is a downdraft system and works really well- we never smell formalin fumes when using that station. I would recommend this unit and when we get to replace our other grossing stations we are most likely to be getting more of the Gross Lab Senior. Hope that helps, Vicki Gauch Albany Medical Center Albany, NY >>> "Histology" 10/1/03 11:04:04 PM >>> Our lab is in the process of purchasing our automated stainer for routine H&E's,Grossing Work Station,Microtom,Cryotom,Embedding center and processor (300 cassettes). I would really appreciate comments from anyone who really likes, or dislikes the machines that they are using. Our plan is to get an automatic coverslipper as well. Thanks in advance!!! Muhammad Tahseen Histology Supervisor Deptt. Pathology Shaukat Khanum Cancer Hospital And Research Center Lahore. Pakistan Ph. +92 42 5180725-36 Ext 2369, Fax. +92 42 5180723 e-mail. Histology@skm.org.pk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Liam.Brennan <@t> bll.n-i.nhs.uk Thu Oct 2 09:10:22 2003 From: Liam.Brennan <@t> bll.n-i.nhs.uk (Brennan, Liam) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Superfrost Plus Slides Message-ID: Dear Bill We have experienced uneven staining on our Ventana NexES stainer over the past number of months. It is not confined to any particular antibody, and does'nt happen all the time. We have been told by Ventana that it is a worldwide problem with coated slides. Apparently a number of brands are manufactured in the same factory. We use Starfrost slides from Knittel Glaser in Germany. As yet, working with the people from Ventana, and trying a variety of slide brands, we have been unable to resolve the problem. I would be interested in hearing from others that have encountered similar problems, and what actions they have taken to resolve the issue. Liam Brennan Histopathology Dept Belfast City Hospital Northern Ireland -----Original Message----- From: Bill Sinai [mailto:bills@icpmr.wsahs.nsw.gov.au] Sent: 01 October 2003 22:31 To: histonet (E-mail) Subject: [Histonet] Superfrost Plus Slides Dear All, Has anyone had a problem with Superfrost Plus slides on automated Immunohistochemistry Staining machines? If so what have the problems been? Also if the problem was resolved and how it was resolved. We obtain our slides through a third party agency in Australia. We are at present using Menzel-Glaser (German) brand. I am led to believe they are the largest slide and coverglass manufacturer in the world. 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Thu Oct 2 09:13:43 2003 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] SAKURA TISSUE -TEK SCA Coverslipper Message-ID: I have used one for years and really like it. I know that others have stated on histonet that they have had problems with the tape coming loose. It is important to dispense an adequate volume of xylene on to the slide. I would say atleast 10 drops of xylene. Daily clean the receiving chute with a little xylene, to remove any resinous residue that could accumulate and impede the slides from entering the receiving basket. It is recommended to file the slides against each other to help keep the coverlip film pressed on. I have had slides sit out for over a year without the film coming loose. I would go with the SCA, it is quick and does a clean job. Fred >>> "Histology" 10/01/03 10:38AM >>> > Dear Histonetters, > I am considering purchasing the SAKURA TISSUE -TEK SCA Coverslipper. > I'd appreciate the hearing about your experiences. > Thanks in advanced, > Muhammad Tahseen Histology Supervisor Deptt. Pathology Shaukat Khanum Cancer Hospital And Research Center Lahore. Pakistan Ph. +92 42 5180725-36 Ext 2369, Fax. +92 42 5180723 e-mail. Histology@skm.org.pk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From trohratnyackhospital <@t> hotmail.com Thu Oct 2 10:00:08 2003 From: trohratnyackhospital <@t> hotmail.com (THERESA ROHR) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] DAKO ER and PR antibodies Message-ID: Hi out there in Histonet land: I am having a problem working up dilutions for DAKO's ER (M7047) and PR (M3569) antibodies. I am using the Envision+ Detection system Suggestions from DAKO were: ER 1:100 to 1:150 with Envision+ and a 20 minute target retrieval PR 1:200 to 1:400 with Envision+ and a 20 minute target retrieval. I've gone less and more and still cannot get any where near the staining on these cases that went out to a IHC Laboratory than I am using as a comparison. Any suggestions would certainly be appreciated. _________________________________________________________________ Frustrated with dial-up? Get high-speed for as low as $29.95/month (depending on the local service providers in your area). https://broadband.msn.com From trohratnyackhospital <@t> hotmail.com Thu Oct 2 10:03:45 2003 From: trohratnyackhospital <@t> hotmail.com (THERESA ROHR) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] DAKO ER and PR antibodies Message-ID: Hi out there in Histonet land: I am having a problem working up dilutions for DAKO's ER (M7047) and PR (M3569) antibodies. I am using the Envision+ Detection system Suggestions from DAKO were: ER 1:100 to 1:150 with Envision+ and a 20 minute target retrieval PR 1:200 to 1:400 with Envision+ and a 20 minute target retrieval. I've gone less and more and still cannot get any where near the staining on these cases that went out to a IHC Laboratory than I am using as a comparison. Any suggestions would certainly be appreciated. Thanks so much, Theresa Rohr, Section Head-Histology Nyack Hospital, New York trohratnyackhospital@hotmail.com _________________________________________________________________ High-speed Internet access as low as $29.95/month (depending on the local service providers in your area). Click here. https://broadband.msn.com From gcallis <@t> montana.edu Thu Oct 2 10:04:35 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] embryo perfusion In-Reply-To: Message-ID: <3.0.6.32.20031002090435.00bd77c0@gemini.msu.montana.edu> It is possible totally block all endogenous peroxidase/pseudoperoxidase using Glucose Oxidase method. You did not say what your enzyme IHC methods was - peroxidase or alkaline phosphatase, I am presuming HRP or you could do the alk Phos method instead of HRP. I will be happy to send (privately) the glucose oxidase method that works for minimally fixed frozen sections (acetone) and PFA or NBF fixed, paraffin embedded tissues. Sectioning a bit thinner, at 5 um may help in getting rid of endogenous peroxidase with less to quench. You always get some shrinkage in paraffin processing (have a publication buried somewhere on this, 25% shrinkage seemed to be normal) - it happens with ALL tissues, delicate embryos often need special schedules to optimize water removal, clearing and paraffin infiltration. It could be that you are overexposing the fixed embryos to alcohols. This can be adjusted for less time to avoid removal of bound water on tissue components rather than just free water found in tissue spaces. Too long in alcohols, xylene, paraffin is too hot and even adding heat to each step of processing of murine tissues contributes to "overprocessing", rather overexposure to alcohols. It very well may NOT be further fixation BY alcohol, but the bound water removal problem. This also leads to poor sectioning, generally rather dry/friable at times. You did not give the processing schedule for these embryos??? At 02:33 PM 10/2/2003 +0200, you wrote: >Hello, > >Need help on PFA perfusion techniques on rat and mice embryos (E18). I've >tried perfusing (intracardiaque) the mothers (10 minutes with PBS/heparine >then 30 minutes with PFA (pump speed of 2ml /min for mice and 25ml/min for >rats), spread the babies out on the table...one sees that the saline >solution does clear the blood, and the PFA does harden them up. I then make >a sagital slice through each embryo and post fix for 24h followed by >paraffin embedding. I then make 7? sagittal slices. But in the stained >sections there are still an abundance of red blood cells that are a big >source of background for IHC, and the tissue seems underfixed because there >is shrinkage (alcohol fixation during embedding). I've heard that one can >perfuse individual embryos via the umbelical cord. Does anyone have a >description of this with a schema or explanation on how to find the right >artery, pump speed and PFA volume. > >thanks for your time, >Nancy Walker >Molecular Biology Scientist > >Sanofi-Synthelbo Research >B.P. 37 Lab?ge Innopole >31676 LABEGE CEDEX FRANCE > >nancy.walker@sanofi-synthelabo.com >tel : (33)561004179? fax :(33)561004001 > > > >_______________________________________________ >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 laurie.colbert <@t> huntingtonhospital.com Thu Oct 2 10:04:59 2003 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] RE: Sakura Tissue Tek SCA Coverslipper Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5BBF2@EXCHANGE1.huntingtonhospital.com> I love this coverslipper! It's easy to use, fast, is low maintenance, slides can be filed right away, and we've had relatively few problems with it. Laurie Colbert Huntington Memorial Hospital Pasadena, CA From ploykasek <@t> phenopath.com Thu Oct 2 10:55:40 2003 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Clones (ER) In-Reply-To: <002601c387bc$c1c1a280$21b6dc0c@insightbb.com> Message-ID: We have a good success with a couple of new clones from Lab Vision (Neomarker antibodies). These clones are rabbit monoclonals. The ER is clone SP1 and the PR clone is SP2. We did an extensive comparison with these new clones against the ER clone 1D5 and the PR clone 636. The new rabbit monoclonals picked more cases than the older clones. It was not a huge difference, but statistically significant. Both of the antigens are sensitive to fixation/processing vagaries. Since we are a reference lab, and receive all kinds of tissue, it is important to us to have clones that will sensitive despite various fixation/processing. These new clones seemed to be more sensitive across all types of tissue. If you are in a lab where you are controlling the tissue fixation/processing, this may not be as much as an issue for you. Please contact me if you need further info. Hope this info helps. Patti Loykasek Phenopath Laboratories Seattle, WA > Hi All: > I have another question from my Pathologist. By the way, the informative > response from my last question, sold him on the value of the histonet. > Question: > Do you prefer on ER Clone? If so, which clone do you prefer and why? For > use on an automated platform which ER clone is best? Also, do you see > significant changes in ER staining from the core breast biopsy to the > excisional breast specimen. How does fixation change the staining results > of your preferred ER clone? Thanks in advance for your response. > Lena > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From ploykasek <@t> phenopath.com Thu Oct 2 11:02:06 2003 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] FisherBiotech MicroProbe Staining Station In-Reply-To: <5.1.1.6.0.20031001085317.00b97df8@itsa.ucsf.edu> Message-ID: Are you going to be using the MicroProbe for ISH? We are using one for ISH & have been very pleased with it. It is certainly cheaper to buy & operate than some of the other automated ISH instruments. It doesn't take much reagent either, if you place your sections on the lower end of the slide. The ProbeOn slides are a little more expensive, but have very good adhesion. If you have any other questions, please contact me. Patti Loykasek PhenoPath Laboratories Seattle, WA> Dear Busy Colleagues, > I would very much appreciate that you could share with us your experience > with MicroProbe staining system from Fisher Scientific. We intend to get > one for a research project processing only 10-20 slides per day. I would > particularly like to know if staining results correlate closer to > conventional manual staining or closer to Techmate system (also a > capillary action system) as well as the overall performance of the MicroProbe. > Thank you very much in advance! > James Guo > ImmunoVision Technologies, Co. > 100 North Hill Drive, #32 > Brisbane, CA 94005 > Tel: 650-320-4622 > Fax: 650-558-9621 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From RITA.ANGEL <@t> UC.EDU Thu Oct 2 11:18:53 2003 From: RITA.ANGEL <@t> UC.EDU (Rita Angel) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] NSH Meeting in Louisville Message-ID: <5.1.0.14.2.20031002121606.00b7ace0@ucmail3.uc.edu> Hi all, One of our sales reps mentioned there is a NSH meeting in Louisville, KY this month. Can anyone give me more info about that and how we can get some registration forms, etc? Thanks for your help, Rita Angel, HT (ASCP) University of Cincinnati From gareth.davis <@t> Vanderbilt.Edu Thu Oct 2 11:20:44 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Need suggestions for secondary antibody Message-ID: <082C721AF78DB34983E8BA2CD085462105C698@mailbe07> Hello all, I'm having a brain cramp. Here's my problem: I am doing a IHC stain on mouse tissue with a monoclonal antibody. I will be using a biotinylated secondary, but what source should I use? My first indication was to use Anti-mouse, but then I came to my senses and realized that wouldn't work. I will be using an ABC kit and DAB. Any suggestions would be appreciated. Thanks, Gareth Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031002/f80f835a/attachment.htm From Jacquie.Mack <@t> CLS.ab.ca Thu Oct 2 11:40:25 2003 From: Jacquie.Mack <@t> CLS.ab.ca (Mack, Jacquie) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] plastination/museum specimens Message-ID: <30C050525B881C4AAFF41E6D16543E6802541BD7@mail3.cls.ab.ca> Does anyone have any websites/references for museum specimens / plastination that they could pass on to me? Greatly appreciated. Jacqueline Mack Tech III Anatomic Pathology Foothills Medical Center Calgary Laboratory Services ph (403) 944-4162 fax (403) 270-4093 e mail: Jacquie.Mack@CLS.ab.ca -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031002/6e38819e/attachment.htm From janis-rodgers <@t> uiowa.edu Thu Oct 2 12:21:15 2003 From: janis-rodgers <@t> uiowa.edu (Rodgers, Janis) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] (no subject) Message-ID: <5D03ED7B9391D4119D9B0008C76B7B24062B0D1B@uihc-mail1.uihc.uiowa.edu> unsubscribe -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031002/2105d6a6/attachment.htm From gcallis <@t> montana.edu Thu Oct 2 12:23:23 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Need suggestions for secondary antibody In-Reply-To: <082C721AF78DB34983E8BA2CD085462105C698@mailbe07> Message-ID: <3.0.6.32.20031002112323.00bc2b48@gemini.msu.montana.edu> Gareth, Your brain is fine! We have excellent luck with goat antiRat F(ab')2, biotin conjugate, adsorbed to mouse from Biosource/TAGO. You can also buy Donkey antiRat F(ab')2 biotinylated, adsorbed to mouse and other species from Jackson Immunoreasearch. We avoid rabbit (sticky bunny!) secondaries, and always use F(ab')2 frag of IgG rather than whole IgG secondaries to avoid fc receptor problems/background staining. These secondaries run around 0.5 mg/ml, diluted to 1:250 - I presume you are using a Rat antimouse monoclonal? If you have a hamster antimouse monoclonal, then BD Pharmingens hamster cocktail of two monoclonals is excellent OR buy a Donkey antiGolden Syrian F(ab')2 biotinylated adsorbed to mouse or Dk antiArmenian Hamster F(ab')2 biotinylated adsorbed to mouse from Jackson Immunoresearch. You will get crisp results with these host anti specific hamster species/Pharm hamster cocktail secondaries compared to a goat antihamster secondary. We experienced very weak, diffuse results with the latter - a horror! Good luck Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From p_bourne_14526 <@t> yahoo.com Thu Oct 2 12:23:56 2003 From: p_bourne_14526 <@t> yahoo.com (Patricia Bourne) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Parvo Controls Message-ID: <20031002172356.38323.qmail@web10009.mail.yahoo.com> Dear all, I am looking for some Parvo Virus Controls. Do anyone have some they would like to share with us? Thanks for the help in advance. Pat --------------------------------- Do you Yahoo!? The New Yahoo! Shopping - with improved product search -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031002/0a85da2b/attachment.htm From dellav <@t> musc.edu Thu Oct 2 12:24:40 2003 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] NSH Meeting in Louisville Message-ID: this link contains everything you ever wanted to know about NSH and the conference http://www.nsh.org/conventions/index.html contact Peggy@nsh.org if you don't find what you need Vinnie Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> Rita Angel 10/02/03 12:18PM >>> Hi all, One of our sales reps mentioned there is a NSH meeting in Louisville, KY this month. Can anyone give me more info about that and how we can get some registration forms, etc? Thanks for your help, Rita Angel, HT (ASCP) University of Cincinnati _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DCoulter <@t> Lifespan.org Thu Oct 2 12:47:41 2003 From: DCoulter <@t> Lifespan.org (Coulter, Diane) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Amyloid control block Message-ID: Does anyone have any amyloid control blocks I could have? Our lab has gram +&-, helico, iron and others to offer in exchange. Thanks in advance, Diane Coulter, Rhode Island Hospital Histology From pruegg <@t> msn.com Thu Oct 2 14:36:30 2003 From: pruegg <@t> msn.com (Patsy Ruegg) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] histogel Message-ID: Has anyone ever tried using something like histogel to orient tissue for frozen sectioning? I am wondering about the cutting and freezing quality of samples held in histogel til firm and then quick frozen in oct? I need to get cross sections of mouse bile ducts. I do this very successfully using histogel and paraffin processing, but I need to get frozen sections of the same for IHC T subset studies. Please advise. Best regards, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 HP-303-644-4538 HF-303-644-3377 hm email pruegg@msn.com wk email pruegg@colobio.com 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. _________________________________________________________________ Get McAfee virus scanning and cleaning of incoming attachments. Get Hotmail Extra Storage! http://join.msn.com/?PAGE=features/es From spoulos <@t> saa.ars.usda.gov Thu Oct 2 14:46:23 2003 From: spoulos <@t> saa.ars.usda.gov (Sylvia Poulos) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] tissue stored in 10% sucrose Message-ID: Hi All, A quick question, We have a visiting scientist who needs to transport tissues stored in 10% sucrose overseas. We would like to ship them but don't know the best way to do so. Ice packs, dry ice, room temp? Also, has anyone had experience with shipping biohazards overseas and what we would need to do? Thoughts on whether it'd be easier to send it with him or ship it separately? As always, thanks for all the help! Sylvia Sylvia P. Poulos USDA-ARS-Animal Physiology Research Unit Athens, GA 30605 706-583-8279 706-542-0399 (fax) From peptolab <@t> hamptons.com Thu Oct 2 14:57:44 2003 From: peptolab <@t> hamptons.com (peptolab) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Retic stain, H2O2 IHC damage,old stains Message-ID: <001901c3891f$7464a410$e176bd18@JEFF> Dick Dapson at Anatech wants all old historical dyes, right? Hydrogen peroxide quenching will digest/dissolve fresh frozen tissue sections just like it does the pus/crust in a wound. I air dry frozen sections for up to an hour u and fix in cold acetone. Your proteinaceous "gentle" H2O2 definately helped. RE: Retic stain Gomori's is really easy to overtone- five minutes in Gold chloride and it seems to me you'd have no fibers stained at all. I use three minutes for the stinky silver. Developing takes no more than 30 seconds with the 20% formalin. Then tone by just dipping the slides in gold chloride for about five seconds or less. It's easy to overdo it. I used to do a shortened, untoned version omitting the gold chloride and you get a lovely gradation of blacks to grays to browns and golden browns in collagen with varying shades according to density of fibrillar collagenization. Reticulin fibres were still black. Nuclei stained too so kernechtrot becomes superfluous. I should write this stain up. Jeff Silverman Path Asst Southside Hospital Bay Shore NY USA From peptolab <@t> hamptons.com Thu Oct 2 15:08:48 2003 From: peptolab <@t> hamptons.com (peptolab) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Don't jump through HIPPA HOOPS Message-ID: <003101c38920$fef03620$e176bd18@JEFF> I belive the consent package that patients get specifies that certain of their personal information will be shared with other healthcare organizations and insurance companies in the course of their diagnosis, treatment and insurance coverage. Don't worry about it. Jeff Silverman Path Asst QIHC Southside Hospital Bay Shore NY USA From peptolab <@t> hamptons.com Thu Oct 2 15:18:46 2003 From: peptolab <@t> hamptons.com (peptolab) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Exempting foriegn bodies Message-ID: <004001c38922$66cab580$e176bd18@JEFF> Your pathologist must take these potential exemptions from gross examination to the hospital's medical board for discussion and approval. If the clinicians are willing to assume any risk arising from failure to document the specimen, something they can document in the chart or op record, and the medical board approves, gross only's can be forgone. Further, certain tissues: hammer toes and bunions, ped tonsils and adenoids, ped hernias, face lift, belly fat, baby foreskins and the dreaded liposuctions, can be exempt from microscopics unless requested by clinician in a specific case. I understand in Great Britain, melanoma reexcisions are sometimes getting gross exam only or maybe one section if the scar is normal grossly. Jeff Silverman Path Asst Southside Hospital Bay Shore NY USA From HornHV <@t> archildrens.org Thu Oct 2 15:25:02 2003 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Don't jump through HIPPA HOOPS Message-ID: Yes, for the treatment and insurance coverage. You cannot use their specimens for research with approval with your IRB. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: peptolab [mailto:peptolab@hamptons.com] Sent: Thursday, October 02, 2003 3:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Don't jump through HIPPA HOOPS I belive the consent package that patients get specifies that certain of their personal information will be shared with other healthcare organizations and insurance companies in the course of their diagnosis, treatment and insurance coverage. Don't worry about it. Jeff Silverman Path Asst QIHC Southside Hospital Bay Shore NY USA _______________________________________________ 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. Arkansas Children's Hospital From gcallis <@t> montana.edu Thu Oct 2 16:16:51 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] histogel, mouse bile duct embed/freeze In-Reply-To: Message-ID: <3.0.6.32.20031002151651.00bc4d20@gemini.msu.montana.edu> You can use OCT and a Tissue Tek plastic cryomold. We freeze lengths of mouse spinal cord by laying them flat on the bottom of a Tissue Tek cryomold (whatever size works best). You can coat the tissue with bit of OCT, lay tissue in bottom of mold, add more OCT and snap freeze but you must hold the mold flat so the bottom remains uncurved, perfectly flat. After freezing, one can pop the block out, orient the frozen tissue block ON END for cross sections, and freeze onto metal chuck in that orientation. More OCT can be added at that time to build up block face, a gradual process to not thaw block. It is more like a double embedding. Our flattest blocks are made by sitting the cryomold with OCT embedded tissue in the bottom of a plastic petri dish that floats on liquid nitrogen. Do not allow Liq N2 to go INSIDE the dish. The dish can sit on a platform immersed in liquid N2 so the dish cannot tip over or allow Liq N2 into the dish. WE get perfect cross sections of spinal cord with this type of embedding and no freezing artifact. At 01:36 PM 10/2/2003 -0600, you wrote: >Has anyone ever tried using something like histogel to orient tissue for >frozen sectioning? I am wondering about the cutting and freezing quality of >samples held in histogel til firm and then quick frozen in oct? I need to >get cross sections of mouse bile ducts. I do this very successfully using >histogel and paraffin processing, but I need to get frozen sections of the >same for IHC T subset studies. >Please advise. >Best regards, >Patsy > > > >Patsy Ruegg, HT(ASCP)QIHC >IHCtech, LLC >Fitzsimmons BioScience Park >12635 Montview Blvd. Suite 216 >Aurora, CO 80010 >P-720-859-4060 >F-720-859-4110 >HP-303-644-4538 >HF-303-644-3377 >hm email pruegg@msn.com >wk email pruegg@colobio.com > >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. > >_________________________________________________________________ >Get McAfee virus scanning and cleaning of incoming attachments. Get Hotmail >Extra Storage! http://join.msn.com/?PAGE=features/es > > >_______________________________________________ >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 RFail <@t> Charleston.net Thu Oct 2 16:41:57 2003 From: RFail <@t> Charleston.net (Rena Fail) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] DAKO ER and PR antibodies In-Reply-To: Message-ID: <021801c3892e$01d9d5a0$b611a6a5@rena> Try 1:10 for ER and 1:150 for PR with HIER pH 6.0 20 minutes steam, 10 min cool down Rena Fail -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of THERESA ROHR Sent: Thursday, October 02, 2003 11:00 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] DAKO ER and PR antibodies Hi out there in Histonet land: I am having a problem working up dilutions for DAKO's ER (M7047) and PR (M3569) antibodies. I am using the Envision+ Detection system Suggestions from DAKO were: ER 1:100 to 1:150 with Envision+ and a 20 minute target retrieval PR 1:200 to 1:400 with Envision+ and a 20 minute target retrieval. I've gone less and more and still cannot get any where near the staining on these cases that went out to a IHC Laboratory than I am using as a comparison. Any suggestions would certainly be appreciated. _________________________________________________________________ Frustrated with dial-up? Get high-speed for as low as $29.95/month (depending on the local service providers in your area). https://broadband.msn.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From georgecole <@t> ev1.net Thu Oct 2 18:56:30 2003 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Gayle callis' methgod of freezing nerve Message-ID: <000001c38940$ce8d39e0$074dbad0@hppav> Histotechs, Gayle Callis' method of freezing nerves is illustrated in the DVD's in the Muscle and Nerve packet I've been send around. It shows a nerve in a flat plastic mold covered with OCT It stresses the fact that the liquid nitrogen must touch the bottom of the mold only. It must not get into the top of the mold. Freezing causes the OCT to contract. Freezing from the bottom like Gayle says ands the DVD shows, keeps the contraction to only a small dent on the surface of the frozen OCT. No problem. This is shown in the DVD. It's easily filled. If the liquid nitrogen gets into the top of the of the mold, the contraction will take place around the nerve making sectioning difficult indeed. Athough DVD One band 9 shows how to patch gaps like these, there's no reason to do so if you freeze the tissue carefully. georgecole@ev1.net -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031002/31d1d025/attachment.htm From roy.ellis <@t> imvs.sa.gov.au Thu Oct 2 19:18:37 2003 From: roy.ellis <@t> imvs.sa.gov.au (Roy Ellis) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] plastination/museum specimens In-Reply-To: <30C050525B881C4AAFF41E6D16543E6802541BD7@mail3.cls.ab.ca> Message-ID: <002701c38943$e3c41b00$a88a140a@imvs.sa.gov.au> Jacquie Try http://www.kfunigraz.ac.at/anawww/plast for plastination technques. Regards Roy mailto:roy.ellis@imvs.sa.gov.au -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Mack, Jacquie Sent: Friday, 3 October 2003 02:10 AM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] plastination/museum specimens Does anyone have any websites/references for museum specimens / plastination that they could pass on to me? Greatly appreciated. Jacqueline Mack Tech III Anatomic Pathology Foothills Medical Center Calgary Laboratory Services ph (403) 944-4162 fax (403) 270-4093 e mail: Jacquie.Mack@CLS.ab.ca -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031003/5cd45cd8/attachment.htm From gale-kleven <@t> uiowa.edu Thu Oct 2 19:38:20 2003 From: gale-kleven <@t> uiowa.edu (Gale A. Kleven) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Freezing & storage of rat embryo head/brain Message-ID: <007d01c38946$a5c73dc0$b2550640@Hydemobil> I am fairly new to histochemical techniques, and have some tissue storage questions. We are freezing whole rat embryo heads/brains on the fast freeze shelf of a cryostat set to -35 C. Then we place the frozen tissue in vac-u-pac bags, and store at -20 C, with the intent of slicing the specimens within a few months. However, I'm now starting a project where I may not be able to slice and process the tissue (for cytochrome oxidase activity in the brain) for about a year. Concerning long term tissue storage: 1. What are the advantages/disadvantages of long term storage in freezers at -20, versus -40, or -80 C? 2. Because I'm already storing the specimens in vacuum sealed bags, would freezing & storing in OCT add any benefit? (I have experienced OCT that has become "gummy" after more than a few days when stored in a Ziploc bag at -20 C, but haven't tried it with the vacuum sealed bags. 3. Would storage at colder temperatures necessitate a different freezing protocol (e.g. slower or faster freezing)? I appreciate any information you can give me on this subject. Gale ______________________________________________________ Gale A. Kleven Department of Psychology University of Iowa http://www.psychology.uiowa.edu/ E11 Seashore Hall Iowa City, IA 52246 email: gale-kleven@uiowa.edu ______________________________________________________ -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031002/13593b46/attachment.htm From lowman034 <@t> yahoo.com Thu Oct 2 22:05:09 2003 From: lowman034 <@t> yahoo.com (Dave Low) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] sperm staining In-Reply-To: <20031001174550.CB22D3D4DF@zeus.mtsac.edu> Message-ID: <20031003030509.41799.qmail@web40910.mail.yahoo.com> Dear Jennifer, We perform a Pap stain over here. Dave Low HT(ASCP)QIHC Elmendorf Med Ctr --- Jmacdona wrote: --------------------------------- Does anyone have a good staining method for humanspermatozoa? -- Jennifer MacDonald --------- Original Message -------- From:"Kathleen Spencer" To: "Noel D. Clark" Cc:Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Perfusiontechniques in mice/rats Date: 01/10/03 07:32 Attached is thecardiac perfusion protocol that we use regularly. Kathleen Spencer HT, ASCPUTHSC Memphis, TN OnWednesday, October 1, 2003, at 07:45 AM, Noel D. Clark wrote: > Washoping to get input on the various perfusion techniques used by > otherresearch labs for rodents. As always, any help is greatly >appreciated. See you inLouisville! > > > > Thanksin advance, > > Noel > > > > > > > > > > Noel Clark, M.A., HTL (ASCP) > >1919 7th Ave South > > Orthopaedic Research Laboratory > >Center for Metabolic Bone Disease > > UniversityofAlabamaatBirmingham> > Birmingham,Alabama35294 > > > _______________________________________________Histonet mailing listHistonet@lists.utsouthwestern.eduhttp://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________ Do you Yahoo!? The New Yahoo! Shopping - with improved product search http://shopping.yahoo.com From docmichel <@t> netbulmail.com Fri Oct 3 01:48:45 2003 From: docmichel <@t> netbulmail.com (izzet oguz) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] cryotechniques Message-ID: <3696.193.255.128.130.1065163725.webmail@mail1.netbulmail.com> we are looking for a textbook or a complete description source of cryotechniques for light microscopy and protocols... do you know a website or a cryo protocol textbook... Thanks in advance.. ?zzet From c.m.vanderloos <@t> amc.uva.nl Fri Oct 3 03:04:51 2003 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] RE: morphology on frozen tissues Message-ID: <6a708a6a0b3c.6a0b3c6a708a@amc.uva.nl> Dear Sun Zhon, Your slides should reach room temperature first (either still in a box or wrapped in plastic) before doing anything. You should prevent them of getting wet after you take them out of the -80 freezer! We have our cryo's packed per two sections, face-to-face in Parafilm with little carton strips between the edges of the two slides. We unwrap them after the sections have reached room temperature. I cannot imagine that your peroxide at this low concentration will do any harm to your sections. The spider web morphology as you describe is probably due to nuclei that had been damaged by osmotic shock or something. There can be numerous reasons for this. One of the causes is for example leaving acetone-fixed cryostat sections too long in distilled water after staining. You better use tap water instead. Hope this helps. Chris van der Loos Dept. of Cardiovascular Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Sun Zhon >Date Wed, 1 Oct 2003 16:44:47 -0700 (PDT) >To Histonet@lists.utsouthwestern.edu >Subject [Histonet] morphology on frozen tissues > >Dear All, >These were the steps I processed my frozen tissues, after sectioning, >air dry tissues for 30 mins fix in acetone for 10 mins at RT air dry >for 30 mins store in -80 freezer > >The next morning before I did my IHC, I took out the fixed frozen >tissues from the -80 freezer and put them directly in PBS without >letting them come to room temperature. > >My tissues looked like spider webs, especially around the edge. >However, in another experiment I used exactly the same tissues and the >tissues looked beautiful. The only difference was that while quenching >the peroxidase, 0.03% H2O2 in PBS was used on the 1st exp (spider web >morphology); 0.03% in 10% horse serum in 3% BSA was used on the 2nd >exp (beautiful morphology). > >Do you think the H2O2 will make such a big difference on the >morphology or there are some other reasons for the spider web effect? >How do you fix and store your frozen tissues? How do you treat the >tissues before you do your IHC exps? > >Thank you very much for your help. > >Sun Zhon From c.m.vanderloos <@t> amc.uva.nl Fri Oct 3 03:35:19 2003 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] RE: need help about frozen bladder sections Message-ID: <6aaa356b0463.6b04636aaa35@amc.uva.nl> Dear Izzet Oguz, A storage temperature of -20C is not cold enought for proper preservation of the tissue morphology and structure. That might be perhaps the reason why your tissue became so fragile. At snap freezing of your tissues the water content will be frozen as amorphous cristals. However, between -1 and -40 water cristals will reshape from amorphous to needles. It is obvious that the needle structures will damage your tissue morphology very badly. At a temperature below -70 this needle forming process cannot take place anymore. Chris van der Loos Dept. of Cardiovascular Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From "izzet oguz" Date Thu, 2 Oct 2003 09:28:51 +0300 (EEST) To Subject [Histonet] need help about frozen bladder sections we took bladder sections from rats about four months ago..first we froze the tissue in liquid nitrogen. Then left them at -20 C deep freeze for 4 months..after that we could not cut the tissues because they became so fragile..Any suggestions about this problem will be greatly appreciated..Thanks.. ?zzet From c.m.vanderloos <@t> amc.uva.nl Fri Oct 3 03:56:18 2003 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] RE: mast cells Message-ID: <6b482a6b1523.6b15236b482a@amc.uva.nl> Hi Noreen, Apart from the histology solutions there is a very good IHC staining for mast cells too. We have been successful using a directly alk. phosphatase labeled anti-tryptase antibody from Chemicon (MAB1222A). Tryptase is an enzyme present in mast cells only. Beautiful staining on both cryostat (1:500, overnight 4C) or FFPE sections (HIER with citrate6.0, 1:500 overnight 4C). For alk. phosphatase chromogen use either New Fuchsin (Dako K0596), Permanent Red (Dako K0695) or Vector Red (SK-5100). Chris van der Loos Dept. of Cardiovascular Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From "Noreen Gilman" Date Wed, 01 Oct 2003 15:00:20 -0400 To Subject [Histonet] mast cells Can anyone recommend a stain for mast cells? The one we do just doesn't seem to be working as well as we'd like it to. thanks noreen Noreen Gilman, BS, HT (ASCP) QIHC Histopathology Supervisor Broward General Medical Center Ft. Lauderdale, FL 33316 From c.m.vanderloos <@t> amc.uva.nl Fri Oct 3 04:08:56 2003 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Re: RE: mast cells Message-ID: <6b24c96b7bc7.6b7bc76b24c9@amc.uva.nl> Hi Carol, I just want to let you know this! Thanks for the info you had send me Chris ----- Original Message ----- From: "C.M. van der Loos" Date: Friday, October 3, 2003 10:56 am Subject: RE: mast cells > Hi Noreen, > Apart from the histology solutions there is a very good IHC > staining for mast cells too. We have been successful using a directly > alk. phosphatase labeled anti-tryptase antibody from Chemicon > (MAB1222A). > Tryptase is an enzyme present in mast cells only. Beautiful staining > on both cryostat (1:500, overnight 4C) or FFPE sections (HIER with > citrate6.0, 1:500 overnight 4C). For alk. phosphatase chromogen > use either New Fuchsin (Dako K0596), Permanent Red (Dako K0695) or > Vector Red (SK-5100). > > Chris van der Loos > Dept. of Cardiovascular Pathology > Academical Medical Center > Amsterdam - The Netherlands > > ----- Original Message ----- > From "Noreen Gilman" > Date Wed, 01 Oct 2003 15:00:20 -0400 > To > Subject [Histonet] mast cells > > Can anyone recommend a stain for mast cells? The one we do just > doesn't > seem to be working as well as we'd like it to. > thanks > noreen > > Noreen Gilman, BS, HT (ASCP) QIHC > Histopathology Supervisor > Broward General Medical Center > Ft. Lauderdale, FL 33316 > > > From ree3 <@t> leicester.ac.uk Fri Oct 3 06:13:16 2003 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] rodent salivary glands Message-ID: Anyone aware of any websites which have photomicrographs of the various rodent salivary glands?? Richard Edwards MRC TOXICOLOGY UNIT LEICESTER..U.K..... From Noel.Clark <@t> ORTHO.UAB.EDU Fri Oct 3 07:20:18 2003 From: Noel.Clark <@t> ORTHO.UAB.EDU (Noel D. Clark) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Thanks for perfusion responses Message-ID: <85F6C7A1330E794DB8540AFD001CC77E0D6717@ROSCO> Just wanted to say thanks to those who responded to my perfusion request. Cheers, Noel Clark, M.A., HTL (ASCP) 1919 7th Ave South Orthopaedic Research Laboratory Center for Metabolic Bone Disease University of Alabama at Birmingham Birmingham, Alabama 35294 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031003/36b55848/attachment.htm From tflore <@t> lsuhsc.edu Fri Oct 3 07:31:08 2003 From: tflore <@t> lsuhsc.edu (Flores, Teresa) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Study group HT/HTL ASCP Registry Message-ID: Our HT/HTL study group (approximately 7 at this time) will meet for the second time on October 25, 2003 (Saturday morning) at LSUHSC, 1901 Perdido Street, New Orleans, LA, 3rd floor, Seminar Room 8 at 8am - Noon. Any questions please call Teresa at (504)568-6042-work / (504) 347-7576 - home or Rosemary at 467-7071 at home. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031003/0efed13c/attachment.htm From Nancy.Walker <@t> sanofi-synthelabo.com Fri Oct 3 08:03:36 2003 From: Nancy.Walker <@t> sanofi-synthelabo.com (Nancy.Walker@sanofi-synthelabo.com) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] =?iso-8859-1?Q?R=E9f=2E_=3A_Re=3A_[Histonet]_embryo_perfusion?= Message-ID: Dear Gayle, I would appreciate your protocole for glucose Oxidase method. Concerning the shrinkage there was a reply which sent me to a link with the "perfusion one" apparatus, with the following info about shrinkage: The shrinkage occurs when a prewash with physiological saline is followed by fixative. The first action of fixative on the cell membrane is to shut off the sodium pump proteins. Sodium rushes into the cell, followed by water to maintain tonicity, and cells swell and expand. Membrane proteins are fixed and crosslinked to neighboring cells in the swollen position. Later, the cells stabilize and contract, and pull other cells with them. The result is gross shrinkage, local distortion and torn membranes with lost cellular contents from some cells. Tissue Shrinkage can be completely prevented if the extracellular fluid with ions is replaced by a nonionic isotonic solution that cannot enter the cells. This can be accomplished by prewash with 5% (isotonic) sucrose in distilled water. Start the prewash at low pressure, and pump up to 300 mmHg to break the blood brain barrier and wash the extracellular fluid (and red blood cells). Switch shortly thereafter to fixative. The perfusion takes the same time as the traditional procedure, and accomplishes the favorable results bulleted above. What do you think about this? After perfusion of the mother I make a sagital incision of each embryo (so the fixative an inclusion infiltrates) then postfix in PFA overnight and 4h in PBS. My inclusion protocole is the same that I use for whole brain from adult rat and mouse: Alcohol 70 - 4h 2x Alcohol 95 - 2h 3x ETOH - 2 h 2x Clearify (xylene substitute) - 2h 3x paraffin - 2h. Thanks again for your advice, Nancy From Nancy.Walker <@t> sanofi-synthelabo.com Fri Oct 3 08:11:46 2003 From: Nancy.Walker <@t> sanofi-synthelabo.com (Nancy.Walker@sanofi-synthelabo.com) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] shrinkage and Perfusion One system Message-ID: Hello, >From the website mentioned in C. Scouten's mail I recuperated this info: The shrinkage occurs when a prewash with physiological saline is followed by fixative. The first action of fixative on the cell membrane is to shut off the sodium pump proteins. Sodium rushes into the cell, followed by water to maintain tonicity, and cells swell and expand. Membrane proteins are fixed and crosslinked to neighboring cells in the swollen position. Later, the cells stabilize and contract, and pull other cells with them. The result is gross shrinkage, local distortion and torn membranes with lost cellular contents from some cells. Tissue Shrinkage can be completely prevented if the extracellular fluid with ions is replaced by a nonionic isotonic solution that cannot enter the cells. This can be accomplished by prewash with 5% (isotonic) sucrose in distilled water. Start the prewash at low pressure, and pump up to 300 mmHg to break the blood brain barrier and wash the extracellular fluid (and red blood cells). Switch shortly thereafter to fixative. The perfusion takes the same time as the traditional procedure, and accomplishes the favorable results bulleted above. So if I'd like to try out the perfusion techniques that are made simple by the Perfusion one system, what would you suggest: first do a physiological saline solution, followed by a prewash with 5% sucrose and then PFA, or go directly to 5% sucrose then PFA? thanks for your advice, Nancy From powell_sa <@t> Mercer.EDU Fri Oct 3 08:41:36 2003 From: powell_sa <@t> Mercer.EDU (Shirley Powell) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] plastination/museum specimens References: <002701c38943$e3c41b00$a88a140a@imvs.sa.gov.au> Message-ID: <001301c389b4$11330220$a6f2acd1@powellsa1> try www.plastination.com as well. Shirley Powell ----- Original Message ----- From: Roy Ellis To: Mack, Jacquie Cc: Histonet Sent: Thursday, October 02, 2003 8:18 PM Subject: RE: [Histonet] plastination/museum specimens Jacquie Try http://www.kfunigraz.ac.at/anawww/plast for plastination technques. Regards Roy mailto:roy.ellis@imvs.sa.gov.au -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Mack, Jacquie Sent: Friday, 3 October 2003 02:10 AM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] plastination/museum specimens Does anyone have any websites/references for museum specimens / plastination that they could pass on to me? Greatly appreciated. Jacqueline Mack Tech III Anatomic Pathology Foothills Medical Center Calgary Laboratory Services ph (403) 944-4162 fax (403) 270-4093 e mail: Jacquie.Mack@CLS.ab.ca -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031003/c62cae53/attachment.htm From gcallis <@t> montana.edu Fri Oct 3 09:36:32 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:59 2005 Subject: further comments on Re: [Histonet] Gayle callis' method of freezing nerve In-Reply-To: <000001c38940$ce8d39e0$074dbad0@hppav> Message-ID: <3.0.6.32.20031003083632.00bcfc10@gemini.msu.montana.edu> Correction, or rather clarification to these comments: George, I do NOT immerse the bottom of the crymold DIRECTLY into pure liquid nitrogen so this direct contact starts freezing the OCT/tissue/mold bottom per your description. The cryomold with OCT embedded tissue is placed in bottom of the EMPTY platic petri dish that has been precooled by FLOATING the dish on a layer of liquid nitrogen - basically the petri dish is canoeing on liquid N2! The plastic dish is not as cold as direct liquid nitrogen contact, acts as a buffer, but still extremely cold to give perfect artifact free freezing. Freezing this way does not allow for any curvature of the plastic cryomold, it stays perfectly flat. We only use Tissue Tek cryomolds as other molds have plastic that is too thick and conducts heat away slower than the thinner Tissue Tek plastic. One joy of this method is once the OCT/tissue in cryomold is sitting in cold petri dish, you can go to the next block immediatetly allowing for a large volume of blocks/collection session. We do not have to hold a cryomold while it freezes. It takes a steady hand to prevent liquid nitrogen from making contact with OCT over the top of cryomold. When the OCT finishes freezing to top of mold with petri dish methods, there are no indentations, just a nice layer of OCT. The petri dish method is not as fast a freezing as you describe although it results in artifact free tissues, and the tidge of slower freezing probably means good interface of tissue to OCT, we never get gaps unless bubbles are not removed. My experience total immersion of OCT embedded tissues into liquid N2 OR embedded in a mold results in cracked OCT, and a horrible block to section. However, you are holding the mold to permit freezing starting at bottom, and as said before - a steady hand helps. Our preferred method of snap freezing is dry ice/isopentane slurry, to do a ton of blocks at one tissue collection session. Only when we need perfectly flat faced blocks is the petri dish canoeing on liquid nitrogen used. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 At 04:56 PM 10/2/2003 -0700, you wrote: > Histotechs, Gayle Callis’ method of freezing nerves is >illustrated in the DVD’s in the Muscle and Nerve packet I’ve >been send around. It shows a nerve in a flat plastic mold covered with OCT >It stresses the fact that the liquid nitrogen must touch the bottom of the >mold only. It must not get into the top of the mold. Freezing causes the >OCT to contract. Freezing from the bottom like Gayle says ands the DVD >shows, keeps the contraction to only a small dent on the surface of the >frozen OCT. No problem. This is shown in the DVD. It’s easily >filled. If the liquid nitrogen gets into the top of the of the mold, the >contraction will take place around the nerve making sectioning difficult >indeed. Athough DVD One band 9 shows how to patch gaps like these, >there’s no reason to do so if you freeze the tissue carefully. >georgecole@ev1.net From ploykasek <@t> phenopath.com Fri Oct 3 11:18:24 2003 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] DAKO ER and PR antibodies In-Reply-To: <021801c3892e$01d9d5a0$b611a6a5@rena> Message-ID: Just fyi. We have used Dako ER-1D5 at 1:200 & PR-636 at 1:250 using a citrate pressure cook pretreatment and Envision+ detection.>I'm not sure what could be giving you problems with your staining. How long is your primary incubation? Detection incubation? Patti Loykasek PhenoPath Laboratories Seattle, WA > Try 1:10 for ER and 1:150 for PR with HIER pH 6.0 20 minutes steam, 10 > min cool down > Rena Fail > > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of THERESA > ROHR > Sent: Thursday, October 02, 2003 11:00 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] DAKO ER and PR antibodies > > > Hi out there in Histonet land: > > I am having a problem working up dilutions for DAKO's ER (M7047) and PR > (M3569) antibodies. > I am using the Envision+ Detection system > > Suggestions from DAKO were: > > ER 1:100 to 1:150 with Envision+ and a 20 minute target retrieval PR > 1:200 to 1:400 with Envision+ and a 20 minute target retrieval. > > I've gone less and more and still cannot get any where near the staining > on > these cases that went out to a IHC Laboratory than I am using as a > comparison. > > Any suggestions would certainly be appreciated. > > _________________________________________________________________ > Frustrated with dial-up? Get high-speed for as low as $29.95/month > (depending on the local service providers in your area). > https://broadband.msn.com > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From lfaison <@t> mail.mcg.edu Fri Oct 3 13:18:42 2003 From: lfaison <@t> mail.mcg.edu (LaTricia Faison) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Histology Services Message-ID: Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. From tissuearray <@t> hotmail.com Fri Oct 3 14:54:25 2003 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Tissue Microarrays and peroxidase staining artifacts Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031003/480b1879/attachment.htm From micro <@t> superlink.net Fri Oct 3 15:22:10 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Histology Services References: Message-ID: <003701c389ec$1a84e4f0$0ecd5cd1@DJ4VDH31> Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From muololi <@t> umdnj.edu Fri Oct 3 15:26:14 2003 From: muololi <@t> umdnj.edu (Lilia Muolo) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Keeping tissues on membrane slides Message-ID: Hi, membrane slides are like plastic wrap over a glass slide, hand glued or you can buy them in a hard plastic cut-out slide with the plastic membrane sealed into it. I'm not sure you can charge the membrane but I don't lay it on the water bath. It seems that the 12 microns that the researcher wanted it cut at was too thick. I have since found out it should be cut at 6 microns for laser cutting and DNA studies. But I will keep your info in mind when I cut with charged slides. Thanks for the tip. Lil Muolo Immuno. Lab Cancer Institute of New Jersey >>> Margaret Horne 10/1/03 10:45:20 AM >>> I don't know about membrane slides , but I had a lot of problems with tissue floating off during IHC. Then someone pointed out that I was using charged slides but laying them out on the side of the water bath. The water bath is grounded and therefore I may have been losing the charge on the slide and it's the charge that keeps ithe tissue on. Stopped touching the slides to the water bath and stopped losing tissue.What can I say... it worked for me. Membrane slides may be very different. I don't know. Good luck, 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 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031003/1b2538c7/attachment.htm From micro <@t> superlink.net Fri Oct 3 15:22:10 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Histology Services References: Message-ID: <003701c389ec$1a84e4f0$0ecd5cd1@DJ4VDH31> Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ckbyrne <@t> exelixis.com Fri Oct 3 15:31:40 2003 From: ckbyrne <@t> exelixis.com (Carrie Kyle-Byrne) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Tissue Microarrays and peroxidase staining artifacts References: Message-ID: <002401c389ed$59bda0c0$860c1dac@ckbyrnewkst> Thom, i beg to differ with your suggestion that the "halo" staining effect is caused by adhesive in the water bath. we use only distilled water in our baths and only plus slides (no additional adhesives) and we still get this effect when staining on an automated stainer. my personal observation has been that this effect is a direct result of the reagent pool receding to the edge of the tissue. i say this for two reasons: 1. i've witnessed the problem with reagent spreading on plus slides on our dako autostainer (even though we use 0.1% tween in our buffer), and 2. we also use the shandon coverplates for manual staining and we never get the "halo" effect on these slides. just my two cents, Carrie Kyle-Byrne, BHS, HT(ASCP) Assoc. Research Scientist II Antibody Core Lab Signal Transduction Research Exelixis, Inc. 170 Harbor Way P.O. Box 511 South San Francisco CA 94083-0511 USA Phone: (1 650) 837-8023 Fax: (1 650) 837-7220 Email: ckbyrne@exelixis.com ________________________________________________________________ This email message 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 email and destroy all copies of the original message. ----- Original Message ----- From: Thom Jensen To: histonet@lists.utsouthwestern.edu Sent: Friday, October 03, 2003 12:54 PM Subject: [Histonet] Tissue Microarrays and peroxidase staining artifacts Hey Histo World, Some people have said that when staining immunoperoxidase using the .6mm array cores, they get dark stained artifacts around the outer surface of each specimen. This also occurs on small biopsy as well. In our lab we have found this happens when using a type of adhesive in the water bath to stick the tissue specimens to the slide. We have eliminated most of this problem by using distilled water in our water baths and only plus charged slides for all our immuno staining. Every once in a while adhesive may be needed like for bloody specimens or for bone marrows. At least that is what we have found in our lab. Any one else have success with other methods of reducing artifacts aournd or in small specimens? For more information on tissue array construction visit: www.arrayworkshop.com ---------------------------------------------------------------------------- -- High-speed Internet access as low as $29.95/month*. Click here. *Depending on the local service providers in your area. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From adford <@t> compuserve.com Fri Oct 3 15:32:20 2003 From: adford <@t> compuserve.com (John Difford) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Re: Disposal of Old Dyestuffs Message-ID: <200310031632_MC3-1-5129-55EE@compuserve.com> To:Margaret A. Rakas, Ph.D. Manager, Inventory & Regulatory Affairs Clark Science Center Smith College Northampton, MA. 01063 Dear Ms Rakas, I understand from your inquiry that you are proposing to dispose of some old dyestuffs which are no longer required by your College. PLEASE DO NOT DO IT! Many old dyes are no longer produced and are almost irreplaceable. They should be preserved by a responsible body in case a new use is found for an old technique in the future and your samples are the only source of the dye needed. Perhaps the National Society for Histotechnology would be willing to hold such samples for the use of posterity? It would at least be an idea to try them. John Difford Histopathology Royal Free Hospital London England, UK From gcallis <@t> montana.edu Fri Oct 3 15:49:40 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Lovely set up for snap freezing In-Reply-To: Message-ID: <3.0.6.32.20031003144940.00bb5f88@gemini.msu.montana.edu> Gail, >From one Gayle to another, a lovely setup, very stable and efficient. Curious though as to why you don't let blocks touch the dry ice? That has never been a worry for us, as they are going from much colder liquid N2 temps to approx -70 to -90C (dry ice) then into -55C freezer. When using dry ice/isopentane slurry we put newly frozen blocks on dry ice to evaporate isopentane fumes before going to -80C freezer. I don't think the dry ice hurts anything - in fact, you would have to use it to ship frozen blocks. Do you get some kind of defect (scratch marks, etc) from putting blocks on dry ice? We have used a solid block of dry ice to snap freeze with plastic cryomolds, OCT embedded tissue, but if the tissue is large, freezing artifact is present i.e. mouse spleen. We do use solid dry ice on occasion to snap freeze extremely tiny NALT, or nasal associated lymphoid tissue from mouse - one of our toughest tissues to dissect out and maintain flat. We never remove the disposable plastic cryomold until just before mounting block on chuck. Obviously your method is very cost effective and you reuse the molds. Thanks for the info, Gayle Callis At 04:17 PM 10/3/2003 -0400, you wrote: >Gayle, > We basically do the same thing you do. We use a stainless steel square >block sitting in the liquid nitrogen. We fill the foam container with >liquid nitrogen covering the s.s.block, this is left for 5-10 minutes with >adding liquid nitrogen as it goes down. Once the s.s.block is cold enough, >the level of liquid nitrogen is kept below the top of the s.s. block. You >can then set your embedding boat on it. We use the Tissue Tek metal >embedding boats and embedding rings with Shandon's M-1. The metal boat >chills and the M-1 freezes fast enough to keep away the freezing artifact. >The s.s. block is stationary and is easy to set the boat onto. All the >surfaces of the block are flat for cutting. It is easy to do and gives great >blocks and tissue. While we are collecting tissue, we pop the block out of >the boat and put the block into a plastic bag in another foam container with >dry ice to keep them frozen until we are done. The blocks never touch the >dry ice and are transferred to a -55 freezer. > Your way is basically the same. I don't see anything wrong with it. > > Gail Macke,HTL > Shriners Burns Hospital--Cincinnati, Ohio > >-----Original Message----- >From: Gayle Callis [mailto:gcallis@montana.edu] >Sent: Friday, October 03, 2003 10:37 AM >To: George Cole; Histonet@lists.utsouthwestern.edu >Subject: further comments on Re: [Histonet] Gayle callis' method of >freezing nerve > > >Correction, or rather clarification to these comments: > >George, > >I do NOT immerse the bottom of the crymold DIRECTLY into pure liquid >nitrogen so this direct contact starts freezing the OCT/tissue/mold bottom >per your description. The cryomold with OCT embedded tissue is placed in >bottom of the EMPTY platic petri dish that has been precooled by FLOATING >the dish on a layer of liquid nitrogen - basically the petri dish is >canoeing on liquid N2! The plastic dish is not as cold as direct liquid >nitrogen contact, acts as a buffer, but still extremely cold to give >perfect artifact free freezing. Freezing this way does not allow for any >curvature of the plastic cryomold, it stays perfectly flat. We only use >Tissue Tek cryomolds as other molds have plastic that is too thick and >conducts heat away slower than the thinner Tissue Tek plastic. One joy of >this method is once the OCT/tissue in cryomold is sitting in cold petri >dish, you can go to the next block immediatetly allowing for a large volume >of blocks/collection session. We do not have to hold a cryomold while it >freezes. It takes a steady hand to prevent liquid nitrogen from making >contact with OCT over the top of cryomold. > >When the OCT finishes freezing to top of mold with petri dish methods, >there are no indentations, just a nice layer of OCT. The petri dish method >is not as fast a freezing as you describe although it results in artifact >free tissues, and the tidge of slower freezing probably means good >interface of tissue to OCT, we never get gaps unless bubbles are not >removed. > >My experience total immersion of OCT embedded tissues into liquid N2 OR >embedded in a mold results in cracked OCT, and a horrible block to section. >However, you are holding the mold to permit freezing starting at bottom, >and as said before - a steady hand helps. > >Our preferred method of snap freezing is dry ice/isopentane slurry, to do a >ton of blocks at one tissue collection session. Only when we need >perfectly flat faced blocks is the petri dish canoeing on liquid nitrogen >used. > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 > > > >At 04:56 PM 10/2/2003 -0700, you wrote: >> Histotechs, Gayle Callis’ method of freezing nerves is >>illustrated in the DVD’s in the Muscle and Nerve packet I’ve >>been send around. It shows a nerve in a flat plastic mold covered with OCT >>It stresses the fact that the liquid nitrogen must touch the bottom of the >>mold only. It must not get into the top of the mold. Freezing causes the >>OCT to contract. Freezing from the bottom like Gayle says ands the DVD >>shows, keeps the contraction to only a small dent on the surface of the >>frozen OCT. No problem. This is shown in the DVD. It’s easily >>filled. If the liquid nitrogen gets into the top of the of the mold, the >>contraction will take place around the nerve making sectioning difficult >>indeed. Athough DVD One band 9 shows how to patch gaps like these, >>there’s no reason to do so if you freeze the tissue carefully. >>georgecole@ev1.net > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet >CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may >contain confidential and privileged information for the use of the >designated recipients. If you are not the intended recipient, (or >authorized to receive for the recipient) you are hereby notified that you >have received this communication in error and that any review, disclosure, >dissemination, distribution or copying of it or its contents is prohibited. >If you have received this communication in error, please destroy all copies >of this communication and any attachments and contact the sender by reply >e-mail or telephone (813) 281-0300. > > > From micro <@t> superlink.net Fri Oct 3 15:22:10 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Histology Services References: Message-ID: <003701c389ec$1a84e4f0$0ecd5cd1@DJ4VDH31> Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From micro <@t> superlink.net Fri Oct 3 15:22:10 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Histology Services References: Message-ID: <003701c389ec$1a84e4f0$0ecd5cd1@DJ4VDH31> Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From KJohnson <@t> med.miami.edu Fri Oct 3 16:29:23 2003 From: KJohnson <@t> med.miami.edu (Johnson, Kevin) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Cell Culture Staining Message-ID: <3132EBE854CF5B4AA15F9BD2F6781C8D03559257@medex09.med.miami.edu> Greetings, all. A researcher just brought me (at Miller Time on a Friday!) two culture flasks of terminally differentiated Sertoli cells and requested H&E staining. Well. Conventional H&E obviously is out, since polystyrene is not compatible with xylene. Further restrictions: the cells cannot be scraped off, they cannot be replated on glass coverslips, and for reasons of her own, they cannot go back into the incubator pending an answer. They now have been fixed in situ with 10% neutral buffered formalin. Does anyone have a protocol for staining these puppies? If not H&E per se, then is there another staining method that might satisfy her? Thanks in advance, Kevin Johnson University of Miami Diabetes Research Institute From gareth.davis <@t> Vanderbilt.Edu Fri Oct 3 17:10:22 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Cell Culture Staining Message-ID: <082C721AF78DB34983E8BA2CD085462105C6A2@mailbe07> When I H&E my frozen sections I don't use Xylene. I merely skip the Xylene and alcohol steps, go straight to dH2O and stain. When I get to Eosin, I just rinse with dH2O and mount coverslips on (if possible) with ageous mounting media. This is probably no help if you have no way to coverslip them. But, this is my suggestion. Gareth Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -----Original Message----- From: Johnson, Kevin [mailto:KJohnson@med.miami.edu] Sent: Fri 10/3/2003 4:29 PM To: 'histonet@lists.utsouthwestern.edu' Cc: Subject: [Histonet] Cell Culture Staining Greetings, all. A researcher just brought me (at Miller Time on a Friday!) two culture flasks of terminally differentiated Sertoli cells and requested H&E staining. Well. Conventional H&E obviously is out, since polystyrene is not compatible with xylene. Further restrictions: the cells cannot be scraped off, they cannot be replated on glass coverslips, and for reasons of her own, they cannot go back into the incubator pending an answer. They now have been fixed in situ with 10% neutral buffered formalin. Does anyone have a protocol for staining these puppies? If not H&E per se, then is there another staining method that might satisfy her? Thanks in advance, Kevin Johnson University of Miami Diabetes Research Institute _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031003/f871b96b/attachment.htm From micro <@t> superlink.net Fri Oct 3 15:22:10 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Histology Services References: Message-ID: <003701c389ec$1a84e4f0$0ecd5cd1@DJ4VDH31> Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From micro <@t> superlink.net Fri Oct 3 15:22:10 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Histology Services References: Message-ID: <003701c389ec$1a84e4f0$0ecd5cd1@DJ4VDH31> Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From micro <@t> superlink.net Fri Oct 3 15:22:10 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Histology Services References: Message-ID: <003701c389ec$1a84e4f0$0ecd5cd1@DJ4VDH31> Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From micro <@t> superlink.net Fri Oct 3 15:22:10 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Histology Services References: Message-ID: <003701c389ec$1a84e4f0$0ecd5cd1@DJ4VDH31> Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From micro <@t> superlink.net Fri Oct 3 15:22:10 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Histology Services References: Message-ID: <003701c389ec$1a84e4f0$0ecd5cd1@DJ4VDH31> Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From micro <@t> superlink.net Fri Oct 3 15:22:10 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Histology Services References: Message-ID: <003701c389ec$1a84e4f0$0ecd5cd1@DJ4VDH31> Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From micro <@t> superlink.net Fri Oct 3 15:22:10 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Histology Services References: Message-ID: <003701c389ec$1a84e4f0$0ecd5cd1@DJ4VDH31> Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From micro <@t> superlink.net Fri Oct 3 15:22:10 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Histology Services References: Message-ID: <003701c389ec$1a84e4f0$0ecd5cd1@DJ4VDH31> Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mucram11 <@t> earthlink.net Sat Oct 4 07:35:28 2003 From: mucram11 <@t> earthlink.net (Pamela Marcum) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Tissue Microarrays and peroxidase staining artifacts Message-ID: <410-220031064123528406@earthlink.net> Carrie, I agree with you. I have had that effect on manual and automated slides when the reagents begin to recede or even evaporate in either method. Many reasons have been given for this effect however, reagents evaporating, spreading too thin or not enough applied were usually the problem when I looked at it very carefully. Since I have not ever used an adhesive in my water bath that was not ever the problem. I can see where it would contribute if the cores are closely placed and the adhesive and water are allowed to dry without fully draining the slide, causing a thickened layer to form at the edges. The adhesive may well stick to the edges regardless of draining or even have some protein in it from the manufacturing process that forms a bond. We have so many ways we can cause ourselves problems with the best intentions to improve our work. Pam Marcum > [Original Message] > From: Carrie Kyle-Byrne > To: Thom Jensen > Cc: Histonet > Date: 10/3/2003 4:34:02 PM > Subject: Re: [Histonet] Tissue Microarrays and peroxidase staining artifacts > > Thom, > > i beg to differ with your suggestion that the "halo" staining effect is > caused by adhesive in the water bath. we use only distilled water in our > baths and only plus slides (no additional adhesives) and we still get this > effect when staining on an automated stainer. my personal observation has > been that this effect is a direct result of the reagent pool receding to the > edge of the tissue. i say this for two reasons: 1. i've witnessed the > problem with reagent spreading on plus slides on our dako autostainer (even > though we use 0.1% tween in our buffer), and 2. we also use the shandon > coverplates for manual staining and we never get the "halo" effect on these > slides. > > just my two cents, > Carrie Kyle-Byrne, BHS, HT(ASCP) > Assoc. Research Scientist II > Antibody Core Lab > Signal Transduction Research > > Exelixis, Inc. > 170 Harbor Way > P.O. Box 511 > South San Francisco > CA 94083-0511 USA > > Phone: (1 650) 837-8023 > Fax: (1 650) 837-7220 > Email: ckbyrne@exelixis.com > > ________________________________________________________________ > This email message 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 email and destroy all copies of the original message. > > ----- Original Message ----- > From: Thom Jensen > To: histonet@lists.utsouthwestern.edu > Sent: Friday, October 03, 2003 12:54 PM > Subject: [Histonet] Tissue Microarrays and peroxidase staining artifacts > > > Hey Histo World, > > Some people have said that when staining immunoperoxidase using the .6mm > array cores, they get dark stained artifacts around the outer surface of > each specimen. This also occurs on small biopsy as well. In our lab we > have found this happens when using a type of adhesive in the water bath to > stick the tissue specimens to the slide. We have eliminated most of this > problem by using distilled water in our water baths and only plus charged > slides for all our immuno staining. Every once in a while adhesive may be > needed like for bloody specimens or for bone marrows. At least that is what > we have found in our lab. > Any one else have success with other methods of reducing artifacts aournd > or in small specimens? > > > > For more information on tissue array construction visit: > www.arrayworkshop.com > > > > > ---------------------------------------------------------------------------- > -- > High-speed Internet access as low as $29.95/month*. Click here. > *Depending on the local service providers in your area. > _______________________________________________ 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 jluis.palazon <@t> icman.csic.es Sat Oct 4 13:38:31 2003 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] culture cells Message-ID: <20031004183831.CF1295BD2E@perceval.uca.es> maybe you can use trypsine to detach the cells, wash the trypsine out with serum or culture media and them concentrate the cells by centrifugating. After you obtain a concentrated suspention you can drop the cells on the slides using a Pasteur pipete, let them dry and fix with formalin or alcohol, and then treat as a conventional slide for H&E. hope this help José Luis From bjeannette <@t> msn.com Sat Oct 4 13:54:39 2003 From: bjeannette <@t> msn.com (BRENDA PERRY) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Fwd: Re: Steiners Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031004/0cf584b6/attachment.htm -------------- next part -------------- An embedded message was scrubbed... From: Roberta Mosedale Subject: Re: Steiners Date: Thu, 07 Aug 2003 12:42:43 -0400 Size: 4036 Url: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031004/0cf584b6/attachment.eml From aem2002 <@t> med.cornell.edu Sat Oct 4 15:04:36 2003 From: aem2002 <@t> med.cornell.edu (April E. Monchik) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] (no subject) Message-ID: <7764E3041B4.AAAD6F8@mail.med.cornell.edu> A non-text attachment was scrubbed... Name: not available Type: multipart/mixed Size: 142 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031004/35e0ba74/attachment.bin From nick.kirk3 <@t> btopenworld.com Sat Oct 4 15:23:28 2003 From: nick.kirk3 <@t> btopenworld.com (Nick Kirk) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] (no subject) In-Reply-To: <7764E3041B4.AAAD6F8@mail.med.cornell.edu> Message-ID: I always use charged slides with frozen sections and they usually stay firmly stuck to the slide. However, it will depend on what tissue you are cutting. Bone and fibrous tissue etc is notorious for falling off and in those circumstances I would use silanised slides (i.e. APES coated). Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of April E. Monchik Sent: 04 October 2003 21:05 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Hi, I'm have a cryo. question. The sections on my slides are falling off when I stain them. Is there a way to save the sections? April _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histon From peoshel <@t> wisc.edu Sat Oct 4 17:38:06 2003 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Lovely set up for snap freezing In-Reply-To: <3.0.6.32.20031003144940.00bb5f88@gemini.msu.montana.edu> References: <3.0.6.32.20031003144940.00bb5f88@gemini.msu.montana.edu> Message-ID: Gayle, Gail, & et al., A couple of things here: First, why dry ice/isopentance slurry? Why not dry ice/ethanol? or acetone? Yes, both are flammable, but much less so (as in not explosive) than isopentane. I've done a fair amount of freeze-substitution with dry ice-ethanol or acetone, and they work well. And they're just as good -- or poor -- for freezing. Which brings up my second point, which some of you may be tired of hearing but ... You do have freezing artifacts. Just because you can't see them doesn't mean the artifacts aren't there and affecting your staining or localizations. The morphology may be perfectly good for light microscopy, and the staining results may be appropriate as well. But. More rigor needs to be used when working with freeze-fixation, and the exact nature of the purpose of the stains or other reactions must be specified. Otherwise the perfectly acceptable morphology may be badly misleading as to the real nature of the stained elements. There. Rant over. Phil >Gail, > >From one Gayle to another, a lovely setup, very stable and efficient. > >Curious though as to why you don't let blocks touch the dry ice? That has >never been a worry for us, as they are going from much colder liquid N2 >temps to approx -70 to -90C (dry ice) then into -55C freezer. When using >dry ice/isopentane slurry we put newly frozen blocks on dry ice to >evaporate isopentane fumes before going to -80C freezer. I don't think the >dry ice hurts anything - in fact, you would have to use it to ship frozen >blocks. > >Do you get some kind of defect (scratch marks, etc) from putting blocks on >dry ice? We have used a solid block of dry ice to snap freeze with plastic >cryomolds, OCT embedded tissue, but if the tissue is large, freezing >artifact is present i.e. mouse spleen. We do use solid dry ice on occasion >to snap freeze extremely tiny NALT, or nasal associated lymphoid tissue >from mouse - one of our toughest tissues to dissect out and maintain flat. > >We never remove the disposable plastic cryomold until just before mounting >block on chuck. Obviously your method is very cost effective and you reuse >the molds. > >Thanks for the info, > >Gayle Callis > >At 04:17 PM 10/3/2003 -0400, you wrote: >>Gayle, >> We basically do the same thing you do. We use a stainless steel square >>block sitting in the liquid nitrogen. We fill the foam container with >>liquid nitrogen covering the s.s.block, this is left for 5-10 minutes with >>adding liquid nitrogen as it goes down. Once the s.s.block is cold enough, >>the level of liquid nitrogen is kept below the top of the s.s. block. You >>can then set your embedding boat on it. We use the Tissue Tek metal >>embedding boats and embedding rings with Shandon's M-1. The metal boat >>chills and the M-1 freezes fast enough to keep away the freezing artifact. >>The s.s. block is stationary and is easy to set the boat onto. All the >>surfaces of the block are flat for cutting. It is easy to do and gives great >>blocks and tissue. While we are collecting tissue, we pop the block out of >>the boat and put the block into a plastic bag in another foam container with >>dry ice to keep them frozen until we are done. The blocks never touch the >>dry ice and are transferred to a -55 freezer. >> Your way is basically the same. I don't see anything wrong with it. >> >> Gail Macke,HTL >> Shriners Burns Hospital--Cincinnati, Ohio >> >>-----Original Message----- >>From: Gayle Callis [mailto:gcallis@montana.edu] >>Sent: Friday, October 03, 2003 10:37 AM >>To: George Cole; Histonet@lists.utsouthwestern.edu >>Subject: further comments on Re: [Histonet] Gayle callis' method of >>freezing nerve >> >> >>Correction, or rather clarification to these comments: >> >>George, >> >>I do NOT immerse the bottom of the crymold DIRECTLY into pure liquid >>nitrogen so this direct contact starts freezing the OCT/tissue/mold bottom >>per your description. The cryomold with OCT embedded tissue is placed in >>bottom of the EMPTY platic petri dish that has been precooled by FLOATING >>the dish on a layer of liquid nitrogen - basically the petri dish is >>canoeing on liquid N2! The plastic dish is not as cold as direct liquid >>nitrogen contact, acts as a buffer, but still extremely cold to give >>perfect artifact free freezing. Freezing this way does not allow for any >>curvature of the plastic cryomold, it stays perfectly flat. We only use >>Tissue Tek cryomolds as other molds have plastic that is too thick and >>conducts heat away slower than the thinner Tissue Tek plastic. One joy of >>this method is once the OCT/tissue in cryomold is sitting in cold petri >>dish, you can go to the next block immediatetly allowing for a large volume >>of blocks/collection session. We do not have to hold a cryomold while it >>freezes. It takes a steady hand to prevent liquid nitrogen from making >>contact with OCT over the top of cryomold. >> >>When the OCT finishes freezing to top of mold with petri dish methods, >>there are no indentations, just a nice layer of OCT. The petri dish method >>is not as fast a freezing as you describe although it results in artifact >>free tissues, and the tidge of slower freezing probably means good >>interface of tissue to OCT, we never get gaps unless bubbles are not >>removed. >> >>My experience total immersion of OCT embedded tissues into liquid N2 OR >>embedded in a mold results in cracked OCT, and a horrible block to section. >>However, you are holding the mold to permit freezing starting at bottom, >>and as said before - a steady hand helps. >> >>Our preferred method of snap freezing is dry ice/isopentane slurry, to do a >>ton of blocks at one tissue collection session. Only when we need >>perfectly flat faced blocks is the petri dish canoeing on liquid nitrogen >>used. >> >>Gayle Callis >>MT,HT,HTL(ASCP) >>Research Histopathology Supervisor >>Veterinary Molecular Biology >>Montana State University - Bozeman >>PO Box 173610 >>Bozeman MT 59717-3610 >> >> >> >>At 04:56 PM 10/2/2003 -0700, you wrote: >>> Histotechs, Gayle Callis’ method of freezing nerves is >>>illustrated in the DVD’s in the Muscle and Nerve packet I’ve >>>been send around. It shows a nerve in a flat plastic mold covered with OCT >>>It stresses the fact that the liquid nitrogen must touch the bottom of the >>>mold only. It must not get into the top of the mold. Freezing causes the >>>OCT to contract. Freezing from the bottom like Gayle says ands the DVD >>>shows, keeps the contraction to only a small dent on the surface of the >>>frozen OCT. No problem. This is shown in the DVD. It’s easily >>>filled. If the liquid nitrogen gets into the top of the of the mold, the >>>contraction will take place around the nerve making sectioning difficult >>>indeed. Athough DVD One band 9 shows how to patch gaps like these, >>>there’s no reason to do so if you freeze the tissue carefully. >>>georgecole@ev1.net >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may >>contain confidential and privileged information for the use of the >>designated recipients. If you are not the intended recipient, (or >>authorized to receive for the recipient) you are hereby notified that you >>have received this communication in error and that any review, disclosure, >>dissemination, distribution or copying of it or its contents is prohibited. >>If you have received this communication in error, please destroy all copies >>of this communication and any attachments and contact the sender by reply >>e-mail or telephone (813) 281-0300. >> >> >> > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From nick.kirk3 <@t> btopenworld.com Sun Oct 5 02:14:14 2003 From: nick.kirk3 <@t> btopenworld.com (Nick Kirk) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Lovely set up for snap freezing In-Reply-To: Message-ID: Interesting thread. I always thought you used isopentane because it had a much lower freezing point (-160C) compared to ethanol (-114.5C) and Acetone (-95.4C). Surely this would have an effect in a dry ice slurry, which would be near the freezing temperature of both Acetone and Ethanol and therefore not be as effective as the isopentane in quenching the tissue due to the ethanol and acetone starting the freeze crystallisation process? Just a thought. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Philip Oshel Sent: 04 October 2003 23:38 To: Histonet@Pathology.swmed.edu Subject: Re: [Histonet] Lovely set up for snap freezing Gayle, Gail, & et al., A couple of things here: First, why dry ice/isopentance slurry? Why not dry ice/ethanol? or acetone? Yes, both are flammable, but much less so (as in not explosive) than isopentane. I've done a fair amount of freeze-substitution with dry ice-ethanol or acetone, and they work well. And they're just as good -- or poor -- for freezing. Which brings up my second point, which some of you may be tired of hearing but ... You do have freezing artifacts. Just because you can't see them doesn't mean the artifacts aren't there and affecting your staining or localizations. The morphology may be perfectly good for light microscopy, and the staining results may be appropriate as well. But. More rigor needs to be used when working with freeze-fixation, and the exact nature of the purpose of the stains or other reactions must be specified. Otherwise the perfectly acceptable morphology may be badly misleading as to the real nature of the stained elements. There. Rant over. Phil >Gail, > >From one Gayle to another, a lovely setup, very stable and efficient. > >Curious though as to why you don't let blocks touch the dry ice? That has >never been a worry for us, as they are going from much colder liquid N2 >temps to approx -70 to -90C (dry ice) then into -55C freezer. When using >dry ice/isopentane slurry we put newly frozen blocks on dry ice to >evaporate isopentane fumes before going to -80C freezer. I don't think the >dry ice hurts anything - in fact, you would have to use it to ship frozen >blocks. > >Do you get some kind of defect (scratch marks, etc) from putting blocks on >dry ice? We have used a solid block of dry ice to snap freeze with plastic >cryomolds, OCT embedded tissue, but if the tissue is large, freezing >artifact is present i.e. mouse spleen. We do use solid dry ice on occasion >to snap freeze extremely tiny NALT, or nasal associated lymphoid tissue >from mouse - one of our toughest tissues to dissect out and maintain flat. > >We never remove the disposable plastic cryomold until just before mounting >block on chuck. Obviously your method is very cost effective and you reuse >the molds. > >Thanks for the info, > >Gayle Callis > >At 04:17 PM 10/3/2003 -0400, you wrote: >>Gayle, >> We basically do the same thing you do. We use a stainless steel square >>block sitting in the liquid nitrogen. We fill the foam container with >>liquid nitrogen covering the s.s.block, this is left for 5-10 minutes with >>adding liquid nitrogen as it goes down. Once the s.s.block is cold enough, >>the level of liquid nitrogen is kept below the top of the s.s. block. You >>can then set your embedding boat on it. We use the Tissue Tek metal >>embedding boats and embedding rings with Shandon's M-1. The metal boat >>chills and the M-1 freezes fast enough to keep away the freezing artifact. >>The s.s. block is stationary and is easy to set the boat onto. All the >>surfaces of the block are flat for cutting. It is easy to do and gives great >>blocks and tissue. While we are collecting tissue, we pop the block out of >>the boat and put the block into a plastic bag in another foam container with >>dry ice to keep them frozen until we are done. The blocks never touch the >>dry ice and are transferred to a -55 freezer. >> Your way is basically the same. I don't see anything wrong with it. >> >> Gail Macke,HTL >> Shriners Burns Hospital--Cincinnati, Ohio >> >>-----Original Message----- >>From: Gayle Callis [mailto:gcallis@montana.edu] >>Sent: Friday, October 03, 2003 10:37 AM >>To: George Cole; Histonet@lists.utsouthwestern.edu >>Subject: further comments on Re: [Histonet] Gayle callis' method of >>freezing nerve >> >> >>Correction, or rather clarification to these comments: >> >>George, >> >>I do NOT immerse the bottom of the crymold DIRECTLY into pure liquid >>nitrogen so this direct contact starts freezing the OCT/tissue/mold bottom >>per your description. The cryomold with OCT embedded tissue is placed in >>bottom of the EMPTY platic petri dish that has been precooled by FLOATING >>the dish on a layer of liquid nitrogen - basically the petri dish is >>canoeing on liquid N2! The plastic dish is not as cold as direct liquid >>nitrogen contact, acts as a buffer, but still extremely cold to give >>perfect artifact free freezing. Freezing this way does not allow for any >>curvature of the plastic cryomold, it stays perfectly flat. We only use >>Tissue Tek cryomolds as other molds have plastic that is too thick and >>conducts heat away slower than the thinner Tissue Tek plastic. One joy of >>this method is once the OCT/tissue in cryomold is sitting in cold petri >>dish, you can go to the next block immediatetly allowing for a large volume >>of blocks/collection session. We do not have to hold a cryomold while it >>freezes. It takes a steady hand to prevent liquid nitrogen from making >>contact with OCT over the top of cryomold. >> >>When the OCT finishes freezing to top of mold with petri dish methods, >>there are no indentations, just a nice layer of OCT. The petri dish method >>is not as fast a freezing as you describe although it results in artifact >>free tissues, and the tidge of slower freezing probably means good >>interface of tissue to OCT, we never get gaps unless bubbles are not >>removed. >> >>My experience total immersion of OCT embedded tissues into liquid N2 OR >>embedded in a mold results in cracked OCT, and a horrible block to section. >>However, you are holding the mold to permit freezing starting at bottom, >>and as said before - a steady hand helps. >> >>Our preferred method of snap freezing is dry ice/isopentane slurry, to do a >>ton of blocks at one tissue collection session. Only when we need >>perfectly flat faced blocks is the petri dish canoeing on liquid nitrogen >>used. >> >>Gayle Callis >>MT,HT,HTL(ASCP) >>Research Histopathology Supervisor >>Veterinary Molecular Biology >>Montana State University - Bozeman >>PO Box 173610 >>Bozeman MT 59717-3610 >> >> >> >>At 04:56 PM 10/2/2003 -0700, you wrote: >>> Histotechs, Gayle Callis’ method of freezing nerves is >>>illustrated in the DVD’s in the Muscle and Nerve packet I’ve >>>been send around. It shows a nerve in a flat plastic mold covered with OCT >>>It stresses the fact that the liquid nitrogen must touch the bottom of the >>>mold only. It must not get into the top of the mold. Freezing causes the >>>OCT to contract. Freezing from the bottom like Gayle says ands the DVD >>>shows, keeps the contraction to only a small dent on the surface of the >>>frozen OCT. No problem. This is shown in the DVD. It’s easily >>>filled. If the liquid nitrogen gets into the top of the of the mold, the >>>contraction will take place around the nerve making sectioning difficult >>>indeed. Athough DVD One band 9 shows how to patch gaps like these, >>>there’s no reason to do so if you freeze the tissue carefully. >>>georgecole@ev1.net >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may >>contain confidential and privileged information for the use of the >>designated recipients. If you are not the intended recipient, (or >>authorized to receive for the recipient) you are hereby notified that you >>have received this communication in error and that any review, disclosure, >>dissemination, distribution or copying of it or its contents is prohibited. >>If you have received this communication in error, please destroy all copies >>of this communication and any attachments and contact the sender by reply >>e-mail or telephone (813) 281-0300. >> >> >> > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From peoshel <@t> wisc.edu Sun Oct 5 08:31:21 2003 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Lovely set up for snap freezing In-Reply-To: <7BA50F61CB491B4692B8BCFBB656B5F924AA81@EXCHCLUSTER01.lj.gnf.org> References: <7BA50F61CB491B4692B8BCFBB656B5F924AA81@EXCHCLUSTER01.lj.gnf.org> Message-ID: Interesting. How can the isopentane be colder than the dry ice? The ethanol or acetone is at dry ice temperature, once equilibrated. Phil >Take temperatures of your slurry, you will find that the temperature >with isopentane is 20-30 degrees colder. > > -----Original Message----- > From: Philip Oshel [mailto:peoshel@wisc.edu] > Sent: Sat 10/4/2003 3:38 PM > To: Histonet@Pathology.swmed.edu > Cc: > Subject: Re: [Histonet] Lovely set up for snap freezing > > > > Gayle, Gail, & et al., > > A couple of things here: > First, why dry ice/isopentance slurry? Why not dry ice/ethanol? or > acetone? Yes, both are flammable, but much less so (as in not > explosive) than isopentane. I've done a fair amount of > freeze-substitution with dry ice-ethanol or acetone, and they work > well. And they're just as good -- or poor -- for freezing. > Which brings up my second point, which some of you may be tired of > hearing but ... > You do have freezing artifacts. Just because you can't see them > doesn't mean the artifacts aren't there and affecting your staining > or localizations. > The morphology may be perfectly good for light microscopy, and the > staining results may be appropriate as well. But. More rigor needs to > be used when working with freeze-fixation, and the exact nature of > the purpose of the stains or other reactions must be specified. > Otherwise the perfectly acceptable morphology may be badly misleading > as to the real nature of the stained elements. > There. Rant over. > > Phil > > >Gail, > > > >From one Gayle to another, a lovely setup, very stable and efficient. > > > >Curious though as to why you don't let blocks touch the dry >ice? That has > >never been a worry for us, as they are going from much >colder liquid N2 > >temps to approx -70 to -90C (dry ice) then into -55C >freezer. When using > >dry ice/isopentane slurry we put newly frozen blocks on dry ice to > >evaporate isopentane fumes before going to -80C freezer. I >don't think the > >dry ice hurts anything - in fact, you would have to use it >to ship frozen > >blocks. > > > >Do you get some kind of defect (scratch marks, etc) from >putting blocks on > >dry ice? We have used a solid block of dry ice to snap >freeze with plastic > >cryomolds, OCT embedded tissue, but if the tissue is large, freezing > >artifact is present i.e. mouse spleen. We do use solid dry >ice on occasion > >to snap freeze extremely tiny NALT, or nasal associated >lymphoid tissue > >from mouse - one of our toughest tissues to dissect out and >maintain flat. > > > >We never remove the disposable plastic cryomold until just >before mounting > >block on chuck. Obviously your method is very cost effective >and you reuse > >the molds. > > > >Thanks for the info, > > > >Gayle Callis > > > >At 04:17 PM 10/3/2003 -0400, you wrote: > >>Gayle, > >> We basically do the same thing you do. We use a >stainless steel square > >>block sitting in the liquid nitrogen. We fill the foam >container with > >>liquid nitrogen covering the s.s.block, this is left for >5-10 minutes with > >>adding liquid nitrogen as it goes down. Once the s.s.block >is cold enough, > >>the level of liquid nitrogen is kept below the top of the >s.s. block. You > >>can then set your embedding boat on it. We use the Tissue Tek metal > >>embedding boats and embedding rings with Shandon's M-1. >The metal boat > >>chills and the M-1 freezes fast enough to keep away the >freezing artifact. > >>The s.s. block is stationary and is easy to set the boat >onto. All the > >>surfaces of the block are flat for cutting. It is easy to >do and gives great > >>blocks and tissue. While we are collecting tissue, we pop >the block out of > >>the boat and put the block into a plastic bag in another >foam container with > >>dry ice to keep them frozen until we are done. The blocks >never touch the > >>dry ice and are transferred to a -55 freezer. > >> Your way is basically the same. I don't see anything >wrong with it. > >> > >> Gail Macke,HTL > >> Shriners Burns Hospital--Cincinnati, Ohio > >> > >>-----Original Message----- > >>From: Gayle Callis [mailto:gcallis@montana.edu] > >>Sent: Friday, October 03, 2003 10:37 AM > >>To: George Cole; Histonet@lists.utsouthwestern.edu > >>Subject: further comments on Re: [Histonet] Gayle callis' method of > >>freezing nerve > >> > >> > >>Correction, or rather clarification to these comments: > >> > >>George, > >> > >>I do NOT immerse the bottom of the crymold DIRECTLY into pure liquid > >>nitrogen so this direct contact starts freezing the >OCT/tissue/mold bottom > >>per your description. The cryomold with OCT embedded >tissue is placed in > >>bottom of the EMPTY platic petri dish that has been >precooled by FLOATING > >>the dish on a layer of liquid nitrogen - basically the petri dish is > >>canoeing on liquid N2! The plastic dish is not as cold as >direct liquid > >>nitrogen contact, acts as a buffer, but still extremely cold to give > >>perfect artifact free freezing. Freezing this way does not >allow for any > >>curvature of the plastic cryomold, it stays perfectly flat. >We only use > >>Tissue Tek cryomolds as other molds have plastic that is >too thick and > >>conducts heat away slower than the thinner Tissue Tek >plastic. One joy of > >>this method is once the OCT/tissue in cryomold is sitting >in cold petri > >>dish, you can go to the next block immediatetly allowing >for a large volume > >>of blocks/collection session. We do not have to hold a >cryomold while it > >>freezes. It takes a steady hand to prevent liquid nitrogen >from making > >>contact with OCT over the top of cryomold. > >> > >>When the OCT finishes freezing to top of mold with petri >dish methods, > >>there are no indentations, just a nice layer of OCT. The >petri dish method > >>is not as fast a freezing as you describe although it >results in artifact > >>free tissues, and the tidge of slower freezing probably means good > >>interface of tissue to OCT, we never get gaps unless bubbles are not > >>removed. > >> > >>My experience total immersion of OCT embedded tissues into >liquid N2 OR > >>embedded in a mold results in cracked OCT, and a horrible >block to section. > >>However, you are holding the mold to permit freezing >starting at bottom, > >>and as said before - a steady hand helps. > >> > >>Our preferred method of snap freezing is dry ice/isopentane >slurry, to do a > >>ton of blocks at one tissue collection session. Only when we need > >>perfectly flat faced blocks is the petri dish canoeing on >liquid nitrogen > >>used. > >> > >>Gayle Callis > >>MT,HT,HTL(ASCP) > >>Research Histopathology Supervisor > >>Veterinary Molecular Biology > >>Montana State University - Bozeman > >>PO Box 173610 > >>Bozeman MT 59717-3610 > >> > >> > >> > >>At 04:56 PM 10/2/2003 -0700, you wrote: > >>> Histotechs, Gayle Callis’ method of >freezing nerves is > >>>illustrated in the DVD’s in the Muscle and Nerve >packet I’ve > >>>been send around. It shows a nerve in a flat plastic mold >covered with OCT > >>>It stresses the fact that the liquid nitrogen must touch >the bottom of the > >>>mold only. It must not get into the top of the mold. >Freezing causes the > >>>OCT to contract. Freezing from the bottom like Gayle says >ands the DVD > >>>shows, keeps the contraction to only a small dent on the >surface of the > >>>frozen OCT. No problem. This is shown in the DVD. >It’s easily > >>>filled. If the liquid nitrogen gets into the top of the of >the mold, the > >>>contraction will take place around the nerve making >sectioning difficult > >>>indeed. Athough DVD One band 9 shows how to patch gaps like these, > >>>there’s no reason to do so if you freeze the tissue carefully. > >>>georgecole@ev1.net > >> > >>_______________________________________________ > >>Histonet mailing list > >>Histonet@lists.utsouthwestern.edu > >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >>CONFIDENTIALITY NOTICE: This e-mail communication and any >attachments may > >>contain confidential and privileged information for the use of the > >>designated recipients. If you are not the intended recipient, (or > >>authorized to receive for the recipient) you are hereby >notified that you > >>have received this communication in error and that any >review, disclosure, > >>dissemination, distribution or copying of it or its >contents is prohibited. > >>If you have received this communication in error, please >destroy all copies > >>of this communication and any attachments and contact the >sender by reply > >>e-mail or telephone (813) 281-0300. > >> > >> > >> > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- > Philip Oshel > Supervisor, AMFSC and BBPIC microscopy facilities > Department of Animal Sciences > University of Wisconsin > 1675 Observatory Drive > Madison, WI 53706 - 1284 > voice: (608) 263-4162 > fax: (608) 262-5157 (dept. fax) > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From peoshel <@t> wisc.edu Sun Oct 5 08:47:09 2003 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Lovely set up for snap freezing In-Reply-To: References: Message-ID: The acetone, ethanol, isopentane, or whatever are going to be as cold as the refrigerant -- dry ice -- and no colder. Perhaps slightly warmer, depending on the set up. Yes, this is closer to the freezing point of EtOH or acetone, but that doesn't matter, they're both still fluid. The best freezing is at liquid nitrogen temperatures, either high-pressure freezing, which gives the greatest depth of evanescent spherule/microcrystaline/amorphous (vitreous) ice, depending who is talking, or propane-jet which is sometimes next best for depth but uses explosive propane under pressure, or plunging into slush nitrogen, which is the coldest, being at the freezing point of nitrogen and gives the best freezing within its depth of good freezing, but gives the shallowest depth of good freezing. I've deliberately not given depths of good freezing because that is highly dependent on the samples, tech headaches, phase of the moon, the usual stuff. Propane jet might work for muscle fibers, HPF would except that it requires very small samples -- sized to fit on a TEM grid -- slush would be good for very small diameter (no, smaller) fibers or the outer 30, maybe 50 microns. Equipment is expensive: HPF very, propane jet, slush relatively cheap. None of these methods use any other vehicle: isopentane, ethane, EtOH, acetone, whatever. Remember: the sample thickness is tissue + carrier medium (buffer, or whatever not blotted off) + sample holder. The less non-tissue the better. Phil >Interesting thread. >I always thought you used isopentane because it had a much lower freezing >point (-160C) compared to ethanol (-114.5C) and Acetone (-95.4C). >Surely this would have an effect in a dry ice slurry, which would be near >the freezing temperature of both Acetone and Ethanol and therefore not be as >effective as the isopentane in quenching the tissue due to the ethanol and >acetone starting the freeze crystallisation process? >Just a thought. > >Nick Kirk >Histopathology >Hinchingbrooke Hospital >Huntingdon >England > >-----Original Message----- >From: histonet-admin@lists.utsouthwestern.edu >[mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Philip >Oshel >Sent: 04 October 2003 23:38 >To: Histonet@Pathology.swmed.edu >Subject: Re: [Histonet] Lovely set up for snap freezing > > >Gayle, Gail, & et al., > >A couple of things here: >First, why dry ice/isopentance slurry? Why not dry ice/ethanol? or >acetone? Yes, both are flammable, but much less so (as in not >explosive) than isopentane. I've done a fair amount of >freeze-substitution with dry ice-ethanol or acetone, and they work >well. And they're just as good -- or poor -- for freezing. >Which brings up my second point, which some of you may be tired of >hearing but ... >You do have freezing artifacts. Just because you can't see them >doesn't mean the artifacts aren't there and affecting your staining >or localizations. >The morphology may be perfectly good for light microscopy, and the >staining results may be appropriate as well. But. More rigor needs to >be used when working with freeze-fixation, and the exact nature of >the purpose of the stains or other reactions must be specified. >Otherwise the perfectly acceptable morphology may be badly misleading >as to the real nature of the stained elements. >There. Rant over. > >Phil > >>Gail, >> >>From one Gayle to another, a lovely setup, very stable and efficient. >> >>Curious though as to why you don't let blocks touch the dry ice? That has >>never been a worry for us, as they are going from much colder liquid N2 >>temps to approx -70 to -90C (dry ice) then into -55C freezer. When using >>dry ice/isopentane slurry we put newly frozen blocks on dry ice to >>evaporate isopentane fumes before going to -80C freezer. I don't think the >>dry ice hurts anything - in fact, you would have to use it to ship frozen >>blocks. >> >>Do you get some kind of defect (scratch marks, etc) from putting blocks on >>dry ice? We have used a solid block of dry ice to snap freeze with plastic >>cryomolds, OCT embedded tissue, but if the tissue is large, freezing >>artifact is present i.e. mouse spleen. We do use solid dry ice on occasion >>to snap freeze extremely tiny NALT, or nasal associated lymphoid tissue >>from mouse - one of our toughest tissues to dissect out and maintain flat. >> >>We never remove the disposable plastic cryomold until just before mounting >>block on chuck. Obviously your method is very cost effective and you reuse >>the molds. >> >>Thanks for the info, >> >>Gayle Callis >> >>At 04:17 PM 10/3/2003 -0400, you wrote: >>>Gayle, >>> We basically do the same thing you do. We use a stainless steel >square >>>block sitting in the liquid nitrogen. We fill the foam container with >>>liquid nitrogen covering the s.s.block, this is left for 5-10 minutes with >>>adding liquid nitrogen as it goes down. Once the s.s.block is cold enough, >>>the level of liquid nitrogen is kept below the top of the s.s. block. You >>>can then set your embedding boat on it. We use the Tissue Tek metal >>>embedding boats and embedding rings with Shandon's M-1. The metal boat >>>chills and the M-1 freezes fast enough to keep away the freezing artifact. >>>The s.s. block is stationary and is easy to set the boat onto. All the >>>surfaces of the block are flat for cutting. It is easy to do and gives >great >>>blocks and tissue. While we are collecting tissue, we pop the block out >of >>>the boat and put the block into a plastic bag in another foam container >with >>>dry ice to keep them frozen until we are done. The blocks never touch the >>>dry ice and are transferred to a -55 freezer. >>> Your way is basically the same. I don't see anything wrong with it. >>> >>> Gail Macke,HTL >>> Shriners Burns Hospital--Cincinnati, Ohio >>> >>>-----Original Message----- >>>From: Gayle Callis [mailto:gcallis@montana.edu] >>>Sent: Friday, October 03, 2003 10:37 AM >>>To: George Cole; Histonet@lists.utsouthwestern.edu >>>Subject: further comments on Re: [Histonet] Gayle callis' method of >>>freezing nerve >>> >>> >>>Correction, or rather clarification to these comments: >>> >>>George, >>> >>>I do NOT immerse the bottom of the crymold DIRECTLY into pure liquid >>>nitrogen so this direct contact starts freezing the OCT/tissue/mold bottom >>>per your description. The cryomold with OCT embedded tissue is placed in >>>bottom of the EMPTY platic petri dish that has been precooled by FLOATING >>>the dish on a layer of liquid nitrogen - basically the petri dish is >>>canoeing on liquid N2! The plastic dish is not as cold as direct liquid >>>nitrogen contact, acts as a buffer, but still extremely cold to give >>>perfect artifact free freezing. Freezing this way does not allow for any >>>curvature of the plastic cryomold, it stays perfectly flat. We only use >>>Tissue Tek cryomolds as other molds have plastic that is too thick and >>>conducts heat away slower than the thinner Tissue Tek plastic. One joy of >>>this method is once the OCT/tissue in cryomold is sitting in cold petri >>>dish, you can go to the next block immediatetly allowing for a large >volume >>>of blocks/collection session. We do not have to hold a cryomold while it >>>freezes. It takes a steady hand to prevent liquid nitrogen from making >>>contact with OCT over the top of cryomold. >>> >>>When the OCT finishes freezing to top of mold with petri dish methods, >>>there are no indentations, just a nice layer of OCT. The petri dish method >>>is not as fast a freezing as you describe although it results in artifact >>>free tissues, and the tidge of slower freezing probably means good >>>interface of tissue to OCT, we never get gaps unless bubbles are not >>>removed. >>> >>>My experience total immersion of OCT embedded tissues into liquid N2 OR >>>embedded in a mold results in cracked OCT, and a horrible block to >section. >>>However, you are holding the mold to permit freezing starting at bottom, >>>and as said before - a steady hand helps. >>> >>>Our preferred method of snap freezing is dry ice/isopentane slurry, to do >a >>>ton of blocks at one tissue collection session. Only when we need >>>perfectly flat faced blocks is the petri dish canoeing on liquid nitrogen >>>used. >>> >>>Gayle Callis >>>MT,HT,HTL(ASCP) >>>Research Histopathology Supervisor >>>Veterinary Molecular Biology >>>Montana State University - Bozeman >>>PO Box 173610 >>>Bozeman MT 59717-3610 >>> >>> >>> >>>At 04:56 PM 10/2/2003 -0700, you wrote: >>>> Histotechs, Gayle Callis’ method of freezing nerves is >>>>illustrated in the DVD’s in the Muscle and Nerve packet I’ve >>>>been send around. It shows a nerve in a flat plastic mold covered with >OCT >>>>It stresses the fact that the liquid nitrogen must touch the bottom of >the >>>>mold only. It must not get into the top of the mold. Freezing causes the >>>>OCT to contract. Freezing from the bottom like Gayle says ands the DVD >>>>shows, keeps the contraction to only a small dent on the surface of the >>>>frozen OCT. No problem. This is shown in the DVD. It’s easily >>>>filled. If the liquid nitrogen gets into the top of the of the mold, the >>>>contraction will take place around the nerve making sectioning difficult >>>>indeed. Athough DVD One band 9 shows how to patch gaps like these, >>>>there’s no reason to do so if you freeze the tissue carefully. >>>>georgecole@ev1.net >>> >>>_______________________________________________ >>>Histonet mailing list >>>Histonet@lists.utsouthwestern.edu >>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>>CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may > >>>contain confidential and privileged information for the use of the >>>designated recipients. If you are not the intended recipient, (or >>>authorized to receive for the recipient) you are hereby notified that you >>>have received this communication in error and that any review, disclosure, >>>dissemination, distribution or copying of it or its contents is >prohibited. >>>If you have received this communication in error, please destroy all >copies >>>of this communication and any attachments and contact the sender by reply >>>e-mail or telephone (813) 281-0300. >>> >>> >>> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >-- >Philip Oshel >Supervisor, AMFSC and BBPIC microscopy facilities >Department of Animal Sciences >University of Wisconsin >1675 Observatory Drive >Madison, WI 53706 - 1284 >voice: (608) 263-4162 >fax: (608) 262-5157 (dept. fax) > >_______________________________________________ >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 -- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From tvedilago <@t> system1.net Sun Oct 5 13:40:07 2003 From: tvedilago <@t> system1.net (Tommy Vedilago) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Histology Temporary positions Message-ID: Hello Histonetters, I am going to begin offering interim or temporary services for Histology labs. Any techs or managers who would like to look into this please give me a call at 866-797-8361 or you can respond via email. Also any of you managers out there who might be in need of interim help please give me a call. There does seem to be a demand for good techs out there on a temporary basis and I will be happy to help in whatever way I can. I will continue to work with permanent placement services as well and I will look forward to hearing any thoughts you all may have on the subject. Sincerely, Tommy Vedilago System 1 Search (864) 627-0012 (864) 627-0013 Fax From histo <@t> skm.org.pk Wed Oct 1 10:23:45 2003 From: histo <@t> skm.org.pk (Histology) Date: Fri Sep 16 15:21:59 2005 Subject: [Histonet] Purchasing machines Grossing Work Station,Microtom etc. Message-ID: Our lab is in the process of purchasing our automated stainer for routine H&E's,Grossing Work Station,Microtom,Cryotom,Embedding center and processor (300 cassettes). I would really appreciate comments from anyone who really likes, or dislikes the machines that they are using. Our plan is to get an automatic coverslipper as well. Thanks in advance!!! Muhammad Tahseen Histology Supervisor Deptt. Pathology Shaukat Khanum Cancer Hospital And Research Center Lahore. Pakistan Ph. +92 42 5180725-36 Ext 2369, Fax. +92 42 5180723 e-mail. Histology@skm.org.pk From Ellin13 <@t> adelphia.net Sun Oct 5 17:46:24 2003 From: Ellin13 <@t> adelphia.net (Jesus Ellin) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Tamtron Training References: <7764E3041B4.AAAD6F8@mail.med.cornell.edu> Message-ID: <001e01c38b92$81c89c40$be284344@yumaaz.adelphia.net.losaca.adelphia.net> Hello there everyone I would like to thank all of you on the help with the kappa and lambda,, but I have another question to ask??? Right now my lab is in the implementation stage for Tamtron and we are going to go live here in about 4 weeks,, Can ANYONE help me with some Ideas for end-user training or can share some ideas of manuals for the end users??? all the help will be greatly appreciated!!! Jesus Ellin HTL ASCP jellin@yumaregional.org From Wijaya.Martanto <@t> chbe.gatech.edu Sun Oct 5 23:27:42 2003 From: Wijaya.Martanto <@t> chbe.gatech.edu (Martanto, Wijaya) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Looking for a hydrophlic dye to use with animal skin in vitro Message-ID: <5D3DB84BB6F5E548AAC9FFEDC310FD091BB8BB@chicken-boo.che.gatech.edu> hi, I was looking for a dye which does not bind locally within animal skin in vitro (tissue marking dye will bind locally within skin). I think the dye should be hydrophilic to allow 'spreading' within the tissue. I am planning to inject the dye into the animal skin in vitro and would like to see the dye distribution within the skin. Currently, I am using tissue marking dye (which I think it's hydrophobic) and it binds locally within the skin (no diffusion/spreading). I was wondering if you have some suggestion on which commercially-available dye I should use. I also have another question: in Conn's 10th Ed., it shows that Aniline Blue has 7% water solubiltiy, however while I looked at older textbooks (Encyclopaedia of Microscopic Stains), it shows that Aniline Blue has 50% water solubility. I would like to hear your opinion about it. Any replies can be directed to my email address shown below: Thanks. Sincerely, Wijaya Martanto wijaya.martanto@chbe.gatech.edu -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031006/23b3f80a/attachment.htm From mparker <@t> epl-inc.com Mon Oct 6 06:02:35 2003 From: mparker <@t> epl-inc.com (Mary Parker) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] unsubscribe Message-ID: <00a801c38bf9$59ba3940$6b01a8c0@93F1H11> Mary Parker, H.T., A.S.C.P. Histology Manager EPL, Inc. P.O. Box 12766 RTP, N.C. 27709 (919) 998-9407 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031006/99ddca0d/attachment.htm From settembr <@t> umdnj.edu Mon Oct 6 06:44:03 2003 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Superfrost Plus Slides Message-ID: No problems with Superfrost Plus Slides. We get them from Fisher Scientific Dana Settembre University Hospital-UMDNJ Newark,NJ USA >>> Bill Sinai 10/1/2003 2:31:29 PM >>> Dear All, Has anyone had a problem with Superfrost Plus slides on automated Immunohistochemistry Staining machines? If so what have the problems been? Also if the problem was resolved and how it was resolved. We obtain our slides through a third party agency in Australia. We are at present using Menzel-Glaser (German) brand. I am led to believe they are the largest slide and coverglass manufacturer in the world. 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo <@t> tiscali.co.uk Mon Oct 6 08:01:53 2003 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Histology Services In-Reply-To: <003701c389ec$1a84e4f0$0ecd5cd1@DJ4VDH31> Message-ID: <002401c38c0a$1cf0f1a0$92362850@KEMLOS> So nice you named it twice, um thrice um....... Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 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-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Markus F. Meyenhofer Sent: 03 October 2003 21:22 To: LaTricia Faison; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histology Services Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ 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 gmacke <@t> shrinenet.org Mon Oct 6 08:13:18 2003 From: gmacke <@t> shrinenet.org (Macke, Gail) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] RE: Lovely set up for snap freezing Message-ID: Gayle, Only reason we don't let the blocks touch the dry ice is sometimes the RA's take awhile to get all the specimens. We found if you leave them sit on the dry ice too long they crack. When in a plastic bag, hanging from the side of the container, not touching the dry ice, we have no cracks in the blocks. Gail Macke -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Friday, October 03, 2003 4:50 PM To: Macke, Gail; Histonet@lists.utsouthwestern.edu Subject: Lovely set up for snap freezing Gail, >From one Gayle to another, a lovely setup, very stable and efficient. Curious though as to why you don't let blocks touch the dry ice? That has never been a worry for us, as they are going from much colder liquid N2 temps to approx -70 to -90C (dry ice) then into -55C freezer. When using dry ice/isopentane slurry we put newly frozen blocks on dry ice to evaporate isopentane fumes before going to -80C freezer. I don't think the dry ice hurts anything - in fact, you would have to use it to ship frozen blocks. Do you get some kind of defect (scratch marks, etc) from putting blocks on dry ice? We have used a solid block of dry ice to snap freeze with plastic cryomolds, OCT embedded tissue, but if the tissue is large, freezing artifact is present i.e. mouse spleen. We do use solid dry ice on occasion to snap freeze extremely tiny NALT, or nasal associated lymphoid tissue from mouse - one of our toughest tissues to dissect out and maintain flat. We never remove the disposable plastic cryomold until just before mounting block on chuck. Obviously your method is very cost effective and you reuse the molds. Thanks for the info, Gayle Callis At 04:17 PM 10/3/2003 -0400, you wrote: >Gayle, > We basically do the same thing you do. We use a stainless steel square >block sitting in the liquid nitrogen. We fill the foam container with >liquid nitrogen covering the s.s.block, this is left for 5-10 minutes with >adding liquid nitrogen as it goes down. Once the s.s.block is cold enough, >the level of liquid nitrogen is kept below the top of the s.s. block. You >can then set your embedding boat on it. We use the Tissue Tek metal >embedding boats and embedding rings with Shandon's M-1. The metal boat >chills and the M-1 freezes fast enough to keep away the freezing artifact. >The s.s. block is stationary and is easy to set the boat onto. All the >surfaces of the block are flat for cutting. It is easy to do and gives great >blocks and tissue. While we are collecting tissue, we pop the block out of >the boat and put the block into a plastic bag in another foam container with >dry ice to keep them frozen until we are done. The blocks never touch the >dry ice and are transferred to a -55 freezer. > Your way is basically the same. I don't see anything wrong with it. > > Gail Macke,HTL > Shriners Burns Hospital--Cincinnati, Ohio > >-----Original Message----- >From: Gayle Callis [mailto:gcallis@montana.edu] >Sent: Friday, October 03, 2003 10:37 AM >To: George Cole; Histonet@lists.utsouthwestern.edu >Subject: further comments on Re: [Histonet] Gayle callis' method of >freezing nerve > > >Correction, or rather clarification to these comments: > >George, > >I do NOT immerse the bottom of the crymold DIRECTLY into pure liquid >nitrogen so this direct contact starts freezing the OCT/tissue/mold bottom >per your description. The cryomold with OCT embedded tissue is placed in >bottom of the EMPTY platic petri dish that has been precooled by FLOATING >the dish on a layer of liquid nitrogen - basically the petri dish is >canoeing on liquid N2! The plastic dish is not as cold as direct liquid >nitrogen contact, acts as a buffer, but still extremely cold to give >perfect artifact free freezing. Freezing this way does not allow for any >curvature of the plastic cryomold, it stays perfectly flat. We only use >Tissue Tek cryomolds as other molds have plastic that is too thick and >conducts heat away slower than the thinner Tissue Tek plastic. One joy of >this method is once the OCT/tissue in cryomold is sitting in cold petri >dish, you can go to the next block immediatetly allowing for a large volume >of blocks/collection session. We do not have to hold a cryomold while it >freezes. It takes a steady hand to prevent liquid nitrogen from making >contact with OCT over the top of cryomold. > >When the OCT finishes freezing to top of mold with petri dish methods, >there are no indentations, just a nice layer of OCT. The petri dish method >is not as fast a freezing as you describe although it results in artifact >free tissues, and the tidge of slower freezing probably means good >interface of tissue to OCT, we never get gaps unless bubbles are not >removed. > >My experience total immersion of OCT embedded tissues into liquid N2 OR >embedded in a mold results in cracked OCT, and a horrible block to section. >However, you are holding the mold to permit freezing starting at bottom, >and as said before - a steady hand helps. > >Our preferred method of snap freezing is dry ice/isopentane slurry, to do a >ton of blocks at one tissue collection session. Only when we need >perfectly flat faced blocks is the petri dish canoeing on liquid nitrogen >used. > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 > > > >At 04:56 PM 10/2/2003 -0700, you wrote: >> Histotechs, Gayle Callis’ method of freezing nerves is >>illustrated in the DVD’s in the Muscle and Nerve packet I’ve >>been send around. It shows a nerve in a flat plastic mold covered with OCT >>It stresses the fact that the liquid nitrogen must touch the bottom of the >>mold only. It must not get into the top of the mold. Freezing causes the >>OCT to contract. Freezing from the bottom like Gayle says ands the DVD >>shows, keeps the contraction to only a small dent on the surface of the >>frozen OCT. No problem. This is shown in the DVD. It’s easily >>filled. If the liquid nitrogen gets into the top of the of the mold, the >>contraction will take place around the nerve making sectioning difficult >>indeed. Athough DVD One band 9 shows how to patch gaps like these, >>there’s no reason to do so if you freeze the tissue carefully. >>georgecole@ev1.net > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet >CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may >contain confidential and privileged information for the use of the >designated recipients. If you are not the intended recipient, (or >authorized to receive for the recipient) you are hereby notified that you >have received this communication in error and that any review, disclosure, >dissemination, distribution or copying of it or its contents is prohibited. >If you have received this communication in error, please destroy all copies >of this communication and any attachments and contact the sender by reply >e-mail or telephone (813) 281-0300. > > > CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. From P.Delahaije <@t> cukz.umcn.nl Mon Oct 6 08:26:38 2003 From: P.Delahaije <@t> cukz.umcn.nl (Delahaije P.) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Aplha B crystalline and pathological conditions Message-ID: <50BC1C1828B0D24EA4001512F7909784012714F4@umcnet14.umcn.nl> Hi! I'm trying to detect alpha B crystalline with an ELISA assay. I have two antibodies (mono and polyklonal) but i havent got a clue about wich buffers to use. I would also like to know if anybody knows something about the pathological conditions alpha b crystalline is produced/overexpressed or used to determine a certain kind of disease? (for instance Alzheimer and so on....) thanks a lot! Peter Delahaije, Laboratory of pediatrics & neurology. Nijmegen Netherlands From cwscouten <@t> myneurolab.com Mon Oct 6 08:53:35 2003 From: cwscouten <@t> myneurolab.com (Charles W. Scouten, Ph.D.) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] RE: shrinkage and Perfusion One system Message-ID: The saline prewash would be just lost time. It accomplishes nothing that will not be accomplished by the sucrose, which is equally effective at washing out the blood (more effective given the high pressure). Start with the sucrose. Time is of the essence, tissue deterioration begins immediately upon anoxia. Switch to fixative when the animal's muscles cease to move. 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 www.myneurolab.com -----Original Message----- From: Nancy.Walker@sanofi-synthelabo.com [mailto:Nancy.Walker@sanofi-synthelabo.com] Sent: Friday, October 03, 2003 8:12 AM To: Charles W. Scouten, Ph.D.; Histonet@lists.utsouthwestern.edu Subject: shrinkage and Perfusion One system Hello, >From the website mentioned in C. Scouten's mail I recuperated this info: The shrinkage occurs when a prewash with physiological saline is followed by fixative. The first action of fixative on the cell membrane is to shut off the sodium pump proteins. Sodium rushes into the cell, followed by water to maintain tonicity, and cells swell and expand. Membrane proteins are fixed and crosslinked to neighboring cells in the swollen position. Later, the cells stabilize and contract, and pull other cells with them. The result is gross shrinkage, local distortion and torn membranes with lost cellular contents from some cells. Tissue Shrinkage can be completely prevented if the extracellular fluid with ions is replaced by a nonionic isotonic solution that cannot enter the cells. This can be accomplished by prewash with 5% (isotonic) sucrose in distilled water. Start the prewash at low pressure, and pump up to 300 mmHg to break the blood brain barrier and wash the extracellular fluid (and red blood cells). Switch shortly thereafter to fixative. The perfusion takes the same time as the traditional procedure, and accomplishes the favorable results bulleted above. So if I'd like to try out the perfusion techniques that are made simple by the Perfusion one system, what would you suggest: first do a physiological saline solution, followed by a prewash with 5% sucrose and then PFA, or go directly to 5% sucrose then PFA? thanks for your advice, Nancy From gcallis <@t> montana.edu Mon Oct 6 09:29:25 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:00 2005 Subject: Why dry ice isopentane slurry for snap freezing? Re: [Histonet] Lovely set up for snap freezing In-Reply-To: References: <3.0.6.32.20031003144940.00bb5f88@gemini.msu.montana.edu> <3.0.6.32.20031003144940.00bb5f88@gemini.msu.montana.edu> Message-ID: <3.0.6.32.20031006082925.00bcfc20@gemini.msu.montana.edu> Simply the if, by accident, the ethanol touches any bare tissues i.e. OCT does not completely cover fresh OR I decide to freeze nonembedded tissue ny this method (undecalcified bone) then I have a fixative in the loop. Isopentane does not fix the tissue, ethanol is a fixative and not always ideal for any immunostaining I might be doing. The same goes for acetone, and when I have grad students who are not as adept at embedding and leave tissue uncovered, then there are potential problems with ethanol/dry ice. At that point, I usually go to pertir dish canoeing on top of liquid nitrogen - no volatile fumes other than good air exchange, don't want to pass out from pure nitrogen in closed spaces. My dry ice/isopentane method is adequate for immunostaining using a light microscope and some other special stains so we live with what we don't see even though the artifact is there. Freeze fixation is what we want to avoid, we prefer to fix AFTER cryosectioning. At 05:38 PM 10/4/2003 -0500, you wrote: >Gayle, Gail, & et al., > >A couple of things here: >First, why dry ice/isopentance slurry? Why not dry ice/ethanol? or >acetone? Yes, both are flammable, but much less so (as in not >explosive) than isopentane. I've done a fair amount of >freeze-substitution with dry ice-ethanol or acetone, and they work >well. And they're just as good -- or poor -- for freezing. >Which brings up my second point, which some of you may be tired of >hearing but ... >You do have freezing artifacts. Just because you can't see them >doesn't mean the artifacts aren't there and affecting your staining >or localizations. >The morphology may be perfectly good for light microscopy, and the >staining results may be appropriate as well. But. More rigor needs to >be used when working with freeze-fixation, and the exact nature of >the purpose of the stains or other reactions must be specified. >Otherwise the perfectly acceptable morphology may be badly misleading >as to the real nature of the stained elements. >There. Rant over. > >Phil > >>Gail, >> >>From one Gayle to another, a lovely setup, very stable and efficient. >> >>Curious though as to why you don't let blocks touch the dry ice? That has >>never been a worry for us, as they are going from much colder liquid N2 >>temps to approx -70 to -90C (dry ice) then into -55C freezer. When using >>dry ice/isopentane slurry we put newly frozen blocks on dry ice to >>evaporate isopentane fumes before going to -80C freezer. I don't think the >>dry ice hurts anything - in fact, you would have to use it to ship frozen >>blocks. >> >>Do you get some kind of defect (scratch marks, etc) from putting blocks on >>dry ice? We have used a solid block of dry ice to snap freeze with plastic >>cryomolds, OCT embedded tissue, but if the tissue is large, freezing >>artifact is present i.e. mouse spleen. We do use solid dry ice on occasion >>to snap freeze extremely tiny NALT, or nasal associated lymphoid tissue >>from mouse - one of our toughest tissues to dissect out and maintain flat. >> >>We never remove the disposable plastic cryomold until just before mounting >>block on chuck. Obviously your method is very cost effective and you reuse >>the molds. >> >>Thanks for the info, >> >>Gayle Callis >> >>At 04:17 PM 10/3/2003 -0400, you wrote: >>>Gayle, >>> We basically do the same thing you do. We use a stainless steel square >>>block sitting in the liquid nitrogen. We fill the foam container with >>>liquid nitrogen covering the s.s.block, this is left for 5-10 minutes with >>>adding liquid nitrogen as it goes down. Once the s.s.block is cold enough, >>>the level of liquid nitrogen is kept below the top of the s.s. block. You >>>can then set your embedding boat on it. We use the Tissue Tek metal >>>embedding boats and embedding rings with Shandon's M-1. The metal boat >>>chills and the M-1 freezes fast enough to keep away the freezing artifact. >>>The s.s. block is stationary and is easy to set the boat onto. All the >>>surfaces of the block are flat for cutting. It is easy to do and gives great >>>blocks and tissue. While we are collecting tissue, we pop the block out of >>>the boat and put the block into a plastic bag in another foam container with >>>dry ice to keep them frozen until we are done. The blocks never touch the >>>dry ice and are transferred to a -55 freezer. >>> Your way is basically the same. I don't see anything wrong with it. >>> >>> Gail Macke,HTL >>> Shriners Burns Hospital--Cincinnati, Ohio >>> >>>-----Original Message----- >>>From: Gayle Callis [mailto:gcallis@montana.edu] >>>Sent: Friday, October 03, 2003 10:37 AM >>>To: George Cole; Histonet@lists.utsouthwestern.edu >>>Subject: further comments on Re: [Histonet] Gayle callis' method of >>>freezing nerve >>> >>> >>>Correction, or rather clarification to these comments: >>> >>>George, >>> >>>I do NOT immerse the bottom of the crymold DIRECTLY into pure liquid >>>nitrogen so this direct contact starts freezing the OCT/tissue/mold bottom >>>per your description. The cryomold with OCT embedded tissue is placed in >>>bottom of the EMPTY platic petri dish that has been precooled by FLOATING >>>the dish on a layer of liquid nitrogen - basically the petri dish is >>>canoeing on liquid N2! The plastic dish is not as cold as direct liquid >>>nitrogen contact, acts as a buffer, but still extremely cold to give >>>perfect artifact free freezing. Freezing this way does not allow for any >>>curvature of the plastic cryomold, it stays perfectly flat. We only use >>>Tissue Tek cryomolds as other molds have plastic that is too thick and >>>conducts heat away slower than the thinner Tissue Tek plastic. One joy of >>>this method is once the OCT/tissue in cryomold is sitting in cold petri >>>dish, you can go to the next block immediatetly allowing for a large volume >>>of blocks/collection session. We do not have to hold a cryomold while it >>>freezes. It takes a steady hand to prevent liquid nitrogen from making >>>contact with OCT over the top of cryomold. >>> >>>When the OCT finishes freezing to top of mold with petri dish methods, >>>there are no indentations, just a nice layer of OCT. The petri dish method >>>is not as fast a freezing as you describe although it results in artifact >>>free tissues, and the tidge of slower freezing probably means good >>>interface of tissue to OCT, we never get gaps unless bubbles are not >>>removed. >>> >>>My experience total immersion of OCT embedded tissues into liquid N2 OR >>>embedded in a mold results in cracked OCT, and a horrible block to section. >>>However, you are holding the mold to permit freezing starting at bottom, >>>and as said before - a steady hand helps. >>> >>>Our preferred method of snap freezing is dry ice/isopentane slurry, to do a >>>ton of blocks at one tissue collection session. Only when we need >>>perfectly flat faced blocks is the petri dish canoeing on liquid nitrogen >>>used. >>> >>>Gayle Callis >>>MT,HT,HTL(ASCP) >>>Research Histopathology Supervisor >>>Veterinary Molecular Biology >>>Montana State University - Bozeman >>>PO Box 173610 >>>Bozeman MT 59717-3610 >>> >>> >>> >>>At 04:56 PM 10/2/2003 -0700, you wrote: >>>> Histotechs, Gayle Callis’ method of freezing nerves is >>>>illustrated in the DVD’s in the Muscle and Nerve packet I’ve >>>>been send around. It shows a nerve in a flat plastic mold covered with OCT >>>>It stresses the fact that the liquid nitrogen must touch the bottom of the >>>>mold only. It must not get into the top of the mold. Freezing causes the >>>>OCT to contract. Freezing from the bottom like Gayle says ands the DVD >>>>shows, keeps the contraction to only a small dent on the surface of the >>>>frozen OCT. No problem. This is shown in the DVD. It’s easily >>>>filled. If the liquid nitrogen gets into the top of the of the mold, the >>>>contraction will take place around the nerve making sectioning difficult >>>>indeed. Athough DVD One band 9 shows how to patch gaps like these, >>>>there’s no reason to do so if you freeze the tissue carefully. >>>>georgecole@ev1.net >>> >>>_______________________________________________ >>>Histonet mailing list >>>Histonet@lists.utsouthwestern.edu >>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>>CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may >>>contain confidential and privileged information for the use of the >>>designated recipients. If you are not the intended recipient, (or >>>authorized to receive for the recipient) you are hereby notified that you >>>have received this communication in error and that any review, disclosure, >>>dissemination, distribution or copying of it or its contents is prohibited. >>>If you have received this communication in error, please destroy all copies >>>of this communication and any attachments and contact the sender by reply >>>e-mail or telephone (813) 281-0300. >>> >>> >>> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >-- >Philip Oshel >Supervisor, AMFSC and BBPIC microscopy facilities >Department of Animal Sciences >University of Wisconsin >1675 Observatory Drive >Madison, WI 53706 - 1284 >voice: (608) 263-4162 >fax: (608) 262-5157 (dept. fax) > >_______________________________________________ >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 Mon Oct 6 09:46:32 2003 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Tissue cores Message-ID: <009801c38c18$a299d480$6401a8c0@instrumedics1> Many labs use the Paraffin Tape-Transfer (PSA) system specifically to ELIMINATE the water bath step which can cause problems. With the PSA the section with 600-1000 cores is captured on a tape and then directly transferred to a slide. The slide is ready for deparaffinazation immediately. Bernice From jlambrey <@t> hotmail.com Mon Oct 6 09:48:02 2003 From: jlambrey <@t> hotmail.com (Julien Lambrey de Souza) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Bond-X autostainer Message-ID: Hi histonetters, Has anyone used the Bond-X immunohistochemical system? Is it any good compared to the Dako for instance? Thanks, Julien. _________________________________________________________________ The new MSN 8: smart spam protection and 2 months FREE* http://join.msn.com/?page=features/junkmail From DPolkovi <@t> PRDUS.JNJ.COM Mon Oct 6 10:11:01 2003 From: DPolkovi <@t> PRDUS.JNJ.COM (Polkovitch, Debbie [PRDUS]) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Keeping bone on a slide Message-ID: <026FB2E8CDEED611BABC00508BAD8AC424D52B@rarusshexs6_ph.prdus.na.jnj.com> Hello, A co-worker of mine is having trouble keeping bone on a positively charged slide during target retrieval using the water bath. Does anyone have any suggestions? Thank you, Deborah Polkovitch, B.S. MT (ASCP) QIHC Research Associate, Drug Discovery, DSE-435 Johnson & Johnson PRD Welsh & McKean Roads, P.O. Box 776 Spring House, PA 19477-0776 Tel : (215) 628-5392 Fax : (215) 540-4887 dpolkovi@prdus.jnj.com -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031006/fb642af7/attachment.htm From mparker <@t> epl-inc.com Mon Oct 6 11:03:32 2003 From: mparker <@t> epl-inc.com (Mary Parker) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] unsubscribe Message-ID: <017901c38c23$648752c0$6b01a8c0@93F1H11> Mary Parker, H.T., A.S.C.P. Histology Manager EPL, Inc. P.O. Box 12766 RTP, N.C. 27709 (919) 998-9407 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031006/48b957d1/attachment.htm From tpmorken <@t> labvision.com Mon Oct 6 11:06:45 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Lovely set up for snap freezing Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CBEE@usca0082k03.rallansci.apogent.com> Dry ice is at -78C, so you're not going to freeze ethanol with that. But the reason to use extremely low temp freezing is to get the fastest RATE of freezing possible to cause the minimum of ice crystal formation. LN2 can cause some problems due to it's propensity to form a vapor layer next to the tissue, and so decrease it's rate of freezing (hence the use of talcum powder on tissue to "breakup" the vapor layer - whether that really happens, I don't know). The other liquids mentioned do not form a vapor layer so may show a higher rate of freezing than LN2 alone. Isopentane freezes around -160C, so using it with LN2 will give a much greater rate of freezing than any dry-ice slush (I always used a digital thermometer to check that the isopentane was at least -150 before freezing tissue). If it's a one-off procedure, then using dry ice slush may be OK, But if you are doing it routinely, LN2/isopentane or using Gayle's method of floating the mold right on the LN2 is the way to go. I've also used the CryoJane (Instrumedics, Inc) apparatus with great results (metal "hammer" in LN2). Tim Morken -----Original Message----- From: Philip Oshel [mailto:peoshel@wisc.edu] Sent: Sunday, October 05, 2003 6:47 AM To: Histonet@Pathology.swmed.edu Subject: RE: [Histonet] Lovely set up for snap freezing The acetone, ethanol, isopentane, or whatever are going to be as cold as the refrigerant -- dry ice -- and no colder. Perhaps slightly warmer, depending on the set up. Yes, this is closer to the freezing point of EtOH or acetone, but that doesn't matter, they're both still fluid. The best freezing is at liquid nitrogen temperatures, either high-pressure freezing, which gives the greatest depth of evanescent spherule/microcrystaline/amorphous (vitreous) ice, depending who is talking, or propane-jet which is sometimes next best for depth but uses explosive propane under pressure, or plunging into slush nitrogen, which is the coldest, being at the freezing point of nitrogen and gives the best freezing within its depth of good freezing, but gives the shallowest depth of good freezing. I've deliberately not given depths of good freezing because that is highly dependent on the samples, tech headaches, phase of the moon, the usual stuff. Propane jet might work for muscle fibers, HPF would except that it requires very small samples -- sized to fit on a TEM grid -- slush would be good for very small diameter (no, smaller) fibers or the outer 30, maybe 50 microns. Equipment is expensive: HPF very, propane jet, slush relatively cheap. None of these methods use any other vehicle: isopentane, ethane, EtOH, acetone, whatever. Remember: the sample thickness is tissue + carrier medium (buffer, or whatever not blotted off) + sample holder. The less non-tissue the better. Phil >Interesting thread. >I always thought you used isopentane because it had a much lower freezing >point (-160C) compared to ethanol (-114.5C) and Acetone (-95.4C). >Surely this would have an effect in a dry ice slurry, which would be near >the freezing temperature of both Acetone and Ethanol and therefore not be as >effective as the isopentane in quenching the tissue due to the ethanol and >acetone starting the freeze crystallisation process? >Just a thought. > >Nick Kirk >Histopathology >Hinchingbrooke Hospital >Huntingdon >England > >-----Original Message----- >From: histonet-admin@lists.utsouthwestern.edu >[mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Philip >Oshel >Sent: 04 October 2003 23:38 >To: Histonet@Pathology.swmed.edu >Subject: Re: [Histonet] Lovely set up for snap freezing > > >Gayle, Gail, & et al., > >A couple of things here: >First, why dry ice/isopentance slurry? Why not dry ice/ethanol? or >acetone? Yes, both are flammable, but much less so (as in not >explosive) than isopentane. I've done a fair amount of >freeze-substitution with dry ice-ethanol or acetone, and they work >well. And they're just as good -- or poor -- for freezing. >Which brings up my second point, which some of you may be tired of >hearing but ... >You do have freezing artifacts. Just because you can't see them >doesn't mean the artifacts aren't there and affecting your staining >or localizations. >The morphology may be perfectly good for light microscopy, and the >staining results may be appropriate as well. But. More rigor needs to >be used when working with freeze-fixation, and the exact nature of >the purpose of the stains or other reactions must be specified. >Otherwise the perfectly acceptable morphology may be badly misleading >as to the real nature of the stained elements. >There. Rant over. > >Phil > >>Gail, >> >>From one Gayle to another, a lovely setup, very stable and efficient. >> >>Curious though as to why you don't let blocks touch the dry ice? That has >>never been a worry for us, as they are going from much colder liquid N2 >>temps to approx -70 to -90C (dry ice) then into -55C freezer. When using >>dry ice/isopentane slurry we put newly frozen blocks on dry ice to >>evaporate isopentane fumes before going to -80C freezer. I don't think the >>dry ice hurts anything - in fact, you would have to use it to ship frozen >>blocks. >> >>Do you get some kind of defect (scratch marks, etc) from putting blocks on >>dry ice? We have used a solid block of dry ice to snap freeze with plastic >>cryomolds, OCT embedded tissue, but if the tissue is large, freezing >>artifact is present i.e. mouse spleen. We do use solid dry ice on occasion >>to snap freeze extremely tiny NALT, or nasal associated lymphoid tissue >>from mouse - one of our toughest tissues to dissect out and maintain flat. >> >>We never remove the disposable plastic cryomold until just before mounting >>block on chuck. Obviously your method is very cost effective and you reuse >>the molds. >> >>Thanks for the info, >> >>Gayle Callis >> >>At 04:17 PM 10/3/2003 -0400, you wrote: >>>Gayle, >>> We basically do the same thing you do. We use a stainless steel >square >>>block sitting in the liquid nitrogen. We fill the foam container with >>>liquid nitrogen covering the s.s.block, this is left for 5-10 minutes with >>>adding liquid nitrogen as it goes down. Once the s.s.block is cold enough, >>>the level of liquid nitrogen is kept below the top of the s.s. block. You >>>can then set your embedding boat on it. We use the Tissue Tek metal >>>embedding boats and embedding rings with Shandon's M-1. The metal boat >>>chills and the M-1 freezes fast enough to keep away the freezing artifact. >>>The s.s. block is stationary and is easy to set the boat onto. All the >>>surfaces of the block are flat for cutting. It is easy to do and gives >great >>>blocks and tissue. While we are collecting tissue, we pop the block out >of >>>the boat and put the block into a plastic bag in another foam container >with >>>dry ice to keep them frozen until we are done. The blocks never touch the >>>dry ice and are transferred to a -55 freezer. >>> Your way is basically the same. I don't see anything wrong with it. >>> >>> Gail Macke,HTL >>> Shriners Burns Hospital--Cincinnati, Ohio >>> >>>-----Original Message----- >>>From: Gayle Callis [mailto:gcallis@montana.edu] >>>Sent: Friday, October 03, 2003 10:37 AM >>>To: George Cole; Histonet@lists.utsouthwestern.edu >>>Subject: further comments on Re: [Histonet] Gayle callis' method of >>>freezing nerve >>> >>> >>>Correction, or rather clarification to these comments: >>> >>>George, >>> >>>I do NOT immerse the bottom of the crymold DIRECTLY into pure liquid >>>nitrogen so this direct contact starts freezing the OCT/tissue/mold bottom >>>per your description. The cryomold with OCT embedded tissue is placed in >>>bottom of the EMPTY platic petri dish that has been precooled by FLOATING >>>the dish on a layer of liquid nitrogen - basically the petri dish is >>>canoeing on liquid N2! The plastic dish is not as cold as direct liquid >>>nitrogen contact, acts as a buffer, but still extremely cold to give >>>perfect artifact free freezing. Freezing this way does not allow for any >>>curvature of the plastic cryomold, it stays perfectly flat. We only use >>>Tissue Tek cryomolds as other molds have plastic that is too thick and >>>conducts heat away slower than the thinner Tissue Tek plastic. One joy of >>>this method is once the OCT/tissue in cryomold is sitting in cold petri >>>dish, you can go to the next block immediatetly allowing for a large >volume >>>of blocks/collection session. We do not have to hold a cryomold while it >>>freezes. It takes a steady hand to prevent liquid nitrogen from making >>>contact with OCT over the top of cryomold. >>> >>>When the OCT finishes freezing to top of mold with petri dish methods, >>>there are no indentations, just a nice layer of OCT. The petri dish method >>>is not as fast a freezing as you describe although it results in artifact >>>free tissues, and the tidge of slower freezing probably means good >>>interface of tissue to OCT, we never get gaps unless bubbles are not >>>removed. >>> >>>My experience total immersion of OCT embedded tissues into liquid N2 OR >>>embedded in a mold results in cracked OCT, and a horrible block to >section. >>>However, you are holding the mold to permit freezing starting at bottom, >>>and as said before - a steady hand helps. >>> >>>Our preferred method of snap freezing is dry ice/isopentane slurry, to do >a >>>ton of blocks at one tissue collection session. Only when we need >>>perfectly flat faced blocks is the petri dish canoeing on liquid nitrogen >>>used. >>> >>>Gayle Callis >>>MT,HT,HTL(ASCP) >>>Research Histopathology Supervisor >>>Veterinary Molecular Biology >>>Montana State University - Bozeman >>>PO Box 173610 >>>Bozeman MT 59717-3610 >>> >>> >>> >>>At 04:56 PM 10/2/2003 -0700, you wrote: >>>> Histotechs, Gayle Callis’ method of freezing nerves is >>>>illustrated in the DVD’s in the Muscle and Nerve packet I’ve >>>>been send around. It shows a nerve in a flat plastic mold covered with >OCT >>>>It stresses the fact that the liquid nitrogen must touch the bottom of >the >>>>mold only. It must not get into the top of the mold. Freezing causes the >>>>OCT to contract. Freezing from the bottom like Gayle says ands the DVD >>>>shows, keeps the contraction to only a small dent on the surface of the >>>>frozen OCT. No problem. This is shown in the DVD. It’s easily >>>>filled. If the liquid nitrogen gets into the top of the of the mold, the >>>>contraction will take place around the nerve making sectioning difficult >>>>indeed. Athough DVD One band 9 shows how to patch gaps like these, >>>>there’s no reason to do so if you freeze the tissue carefully. >>>>georgecole@ev1.net >>> >>>_______________________________________________ >>>Histonet mailing list >>>Histonet@lists.utsouthwestern.edu >>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>>CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may > >>>contain confidential and privileged information for the use of the >>>designated recipients. If you are not the intended recipient, (or >>>authorized to receive for the recipient) you are hereby notified that you >>>have received this communication in error and that any review, disclosure, >>>dissemination, distribution or copying of it or its contents is >prohibited. >>>If you have received this communication in error, please destroy all >copies >>>of this communication and any attachments and contact the sender by reply >>>e-mail or telephone (813) 281-0300. >>> >>> >>> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >-- >Philip Oshel >Supervisor, AMFSC and BBPIC microscopy facilities >Department of Animal Sciences >University of Wisconsin >1675 Observatory Drive >Madison, WI 53706 - 1284 >voice: (608) 263-4162 >fax: (608) 262-5157 (dept. fax) > >_______________________________________________ >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 -- Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tissuearray <@t> hotmail.com Mon Oct 6 11:41:59 2003 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Tissue Cores Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031006/ca0ea2e4/attachment.htm From histobabs <@t> hotmail.com Mon Oct 6 11:47:05 2003 From: histobabs <@t> hotmail.com (Barbara Terrett) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] AMACR/p63 Message-ID: Hi, Is anyone using a AMACR/p63 cocktail antibody, possibly called PIN cocktail? If so , who is your supplier? Tahnks, b _________________________________________________________________ MSN 8 with e-mail virus protection service: 2 months FREE* http://join.msn.com/?page=features/virus From gareth.davis <@t> Vanderbilt.Edu Mon Oct 6 11:50:10 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Advice on cutting Olfactory sections Message-ID: <082C721AF78DB34983E8BA2CD085462105C6A7@mailbe07> I am applying for part-time histology job where I will be cutting paraffin sections of the olfactory area of mice. I've never cut decalcified bone before. I have cut tissue where I just allow them to float on puddles of warm water (while on slide warmer) and other tissue where I allow them to float in a waterbath before placing on a slide. But these have to be serial sections, so I don't see them being floating sections. Anyway, anyone's expertise and suggestions would be greatly appreciated. Thanks Gareth Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031006/f4434c30/attachment.htm From settembr <@t> umdnj.edu Mon Oct 6 11:56:42 2003 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] DAKO ER and PR antibodies Message-ID: Hello Theresa, I use the same ER and PR antibodies that you use and I also use Dako's Target Retrieval Solution My method works well: TRS for 40 minutes in a steamer ER used at 1:20 and incubated for 30 minutes (more can't hurt, up to 60 minutes) PR used at 1:30 and incubated for 20 minutes (more can' hurt either) I use Dako's LSAB2 kit (which is less sensitive than your Envision +, I might go that route and dilute my Ab's) Good Luck Dana Settembre University Hospital-UMDNJ Newark, NJ >>> THERESA ROHR 10/2/2003 8:00:08 AM >>> Hi out there in Histonet land: I am having a problem working up dilutions for DAKO's ER (M7047) and PR (M3569) antibodies. I am using the Envision+ Detection system Suggestions from DAKO were: ER 1:100 to 1:150 with Envision+ and a 20 minute target retrieval PR 1:200 to 1:400 with Envision+ and a 20 minute target retrieval. I've gone less and more and still cannot get any where near the staining on these cases that went out to a IHC Laboratory than I am using as a comparison. Any suggestions would certainly be appreciated. _________________________________________________________________ Frustrated with dial-up? Get high-speed for as low as $29.95/month (depending on the local service providers in your area). https://broadband.msn.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From settembr <@t> umdnj.edu Mon Oct 6 12:03:06 2003 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] BRST2 Message-ID: Hello Paula, I use Brst-2 from Signet cat.# 611-23 It is ready to use but I dilute it further @ a 1:1 dilution and incubate for 15 minutes I pretreat for 5 minutes @ room temperature with Dako's Prot. K cat.# 3020 (they have small and large volume) The detection system that I use is Dako's LSAB2 kit, but other kits work just as well. Good Luck Dana Settembre University Hospital-UMDNJ Newark, NJ >>> Paula Wilder 10/1/2003 4:39:15 PM >>> Can anyone tell me how they handle this antibody? We are getting quite a mixed response from neighboring institutions. Thanks so much! Paula Wilder St. Joseph Medical Center 7601 Osler Drive Towson, MD. _________________________________________________________________ Get McAfee virus scanning and cleaning of incoming attachments. Get Hotmail Extra Storage! http://join.msn.com/?PAGE=features/es _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pzeitlow <@t> bbpllab.com Mon Oct 6 12:01:43 2003 From: pzeitlow <@t> bbpllab.com (Pat Zeitlow) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] AMACR/p63 Message-ID: <813FB33DA405334F947F8BFC6EBD0B2A0BE867@bbplsrv1.bbpl> I use PIN cocktail provided by Biocare Medical. I have had no problems with the antibody and am seeing an increase in orders for it. Pat -----Original Message----- From: Barbara Terrett [mailto:histobabs@hotmail.com] Sent: Monday, October 06, 2003 11:47 AM To: histonet@pathology.swmed.edu Subject: [Histonet] AMACR/p63 Hi, Is anyone using a AMACR/p63 cocktail antibody, possibly called PIN cocktail? If so , who is your supplier? Tahnks, b _________________________________________________________________ MSN 8 with e-mail virus protection service: 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 cfavara <@t> niaid.nih.gov Mon Oct 6 12:14:31 2003 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Superfrost Plus Slides Message-ID: I would be much more inclined to think there is something amiss with the vortex mixers and or the rinses on the Nexus. I use the Nexus and am generally pleased with the results for high volumes but from my experience one clogged vortex mixer can cause erratic and uneven staining. One way to access this would be to run duplicates staining by hand and see if you get the same variation with the slides. I do think in terms quality of staining that staining by hand when done consistently beats any machine hands down!!!! or up!!!!! c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: Brennan, Liam [mailto:Liam.Brennan@bll.n-i.nhs.uk] Sent: Thursday, October 02, 2003 8:10 AM To: 'Bill Sinai'; 'HISTONET' Subject: RE: [Histonet] Superfrost Plus Slides Dear Bill We have experienced uneven staining on our Ventana NexES stainer over the past number of months. It is not confined to any particular antibody, and does'nt happen all the time. We have been told by Ventana that it is a worldwide problem with coated slides. Apparently a number of brands are manufactured in the same factory. We use Starfrost slides from Knittel Glaser in Germany. As yet, working with the people from Ventana, and trying a variety of slide brands, we have been unable to resolve the problem. I would be interested in hearing from others that have encountered similar problems, and what actions they have taken to resolve the issue. Liam Brennan Histopathology Dept Belfast City Hospital Northern Ireland -----Original Message----- From: Bill Sinai [mailto:bills@icpmr.wsahs.nsw.gov.au] Sent: 01 October 2003 22:31 To: histonet (E-mail) Subject: [Histonet] Superfrost Plus Slides Dear All, Has anyone had a problem with Superfrost Plus slides on automated Immunohistochemistry Staining machines? If so what have the problems been? Also if the problem was resolved and how it was resolved. We obtain our slides through a third party agency in Australia. We are at present using Menzel-Glaser (German) brand. I am led to believe they are the largest slide and coverglass manufacturer in the world. 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. _______________________________________________ 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 cfavara <@t> niaid.nih.gov Mon Oct 6 12:30:48 2003 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Cell Culture Staining Message-ID: Why do you need xylene? they are not in paraffin? c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: Johnson, Kevin [mailto:KJohnson@med.miami.edu] Sent: Friday, October 03, 2003 3:29 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Cell Culture Staining Greetings, all. A researcher just brought me (at Miller Time on a Friday!) two culture flasks of terminally differentiated Sertoli cells and requested H&E staining. Well. Conventional H&E obviously is out, since polystyrene is not compatible with xylene. Further restrictions: the cells cannot be scraped off, they cannot be replated on glass coverslips, and for reasons of her own, they cannot go back into the incubator pending an answer. They now have been fixed in situ with 10% neutral buffered formalin. Does anyone have a protocol for staining these puppies? If not H&E per se, then is there another staining method that might satisfy her? Thanks in advance, Kevin Johnson University of Miami Diabetes Research Institute _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dbpiontek <@t> hsc.wvu.edu Mon Oct 6 12:59:49 2003 From: dbpiontek <@t> hsc.wvu.edu (Denise Bland-Piontek) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] COX 1&2 Message-ID: Is anyone currently doing antibodies Cox1 & Cox2? I'm trying to locate a vendor that has a reliable antibody for IHC. So far the only manufacturers I've located don't seem to really have IHC as a priority. As always any help is welcomed! Denise Bland-Piontek WVU From peoshel <@t> wisc.edu Mon Oct 6 13:23:07 2003 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Re: Why dry ice isopentane slurry for snap freezing? In-Reply-To: <3.0.6.32.20031006082925.00bcfc20@gemini.msu.montana.edu> References: <3.0.6.32.20031003144940.00bb5f88@gemini.msu.montana.edu> <3.0.6.32.20031003144940.00bb5f88@gemini.msu.montana.edu> <3.0.6.32.20031006082925.00bcfc20@gemini.msu.montana.edu> Message-ID: OK, I understand this. But, does ethanol or acetone at -78C really act fast enough to coagulate proteins in a few hours? I wouldn't've suspected that. Freeze substitution takes days. I don't know. How thick a layer of OCT do you use? But, a couple of points for the discussion: First, you are freeze-fixing. It's reversible by thawing, but it's still fixation, just physical instead of chemical. Which, from what you are saying, is an important difference. Second, my point with the artifact comment is that invisible artifacts -- i.e., below the resolution of the imaging technique used -- must still be considered. For instance, if you are interested in an enzyme that occurs inside smooth ER at some stage and moves outside the ER at another stage, but the freezing method, or subsequent handling (e.g. warming the tissue to -40C or higher, which is above the recrystallization point) disrupts the ER, then enzyme within the ER can -- almost certainly will -- move outside the ER. This movement may also be subresolution, but if the example enzyme is not accessible to the stain or Ab when within the ER, then you have a false positive, no matter how good the morphology. Phil >Simply the if, by accident, the ethanol touches any bare tissues i.e. OCT >does not completely cover fresh OR I decide to freeze nonembedded tissue >ny this method (undecalcified bone) then I have a fixative in the loop. >Isopentane does not fix the tissue, ethanol is a fixative and not always >ideal for any immunostaining I might be doing. > >The same goes for acetone, and when I have grad students who are not as >adept at embedding and leave tissue uncovered, then there are potential >problems with ethanol/dry ice. At that point, I usually go to pertir dish >canoeing on top of liquid nitrogen - no volatile fumes other than good air >exchange, don't want to pass out from pure nitrogen in closed spaces. > >My dry ice/isopentane method is adequate for immunostaining using a light >microscope and some other special stains so we live with what we don't see >even though the artifact is there. Freeze fixation is what we want to >avoid, we prefer to fix AFTER cryosectioning. > > > >At 05:38 PM 10/4/2003 -0500, you wrote: >>Gayle, Gail, & et al., >> >>A couple of things here: >>First, why dry ice/isopentance slurry? Why not dry ice/ethanol? or >>acetone? Yes, both are flammable, but much less so (as in not >>explosive) than isopentane. I've done a fair amount of >>freeze-substitution with dry ice-ethanol or acetone, and they work >>well. And they're just as good -- or poor -- for freezing. >>Which brings up my second point, which some of you may be tired of >>hearing but ... >>You do have freezing artifacts. Just because you can't see them >>doesn't mean the artifacts aren't there and affecting your staining >>or localizations. >>The morphology may be perfectly good for light microscopy, and the >>staining results may be appropriate as well. But. More rigor needs to >>be used when working with freeze-fixation, and the exact nature of >>the purpose of the stains or other reactions must be specified. >>Otherwise the perfectly acceptable morphology may be badly misleading >>as to the real nature of the stained elements. >>There. Rant over. >> >>Phil >> >>>Gail, >>> >>>From one Gayle to another, a lovely setup, very stable and efficient. >>> >>>Curious though as to why you don't let blocks touch the dry ice? That has >>>never been a worry for us, as they are going from much colder liquid N2 >>>temps to approx -70 to -90C (dry ice) then into -55C freezer. When using >>>dry ice/isopentane slurry we put newly frozen blocks on dry ice to >>>evaporate isopentane fumes before going to -80C freezer. I don't think the >>>dry ice hurts anything - in fact, you would have to use it to ship frozen >>>blocks. >>> >>>Do you get some kind of defect (scratch marks, etc) from putting blocks on >>>dry ice? We have used a solid block of dry ice to snap freeze with plastic >>>cryomolds, OCT embedded tissue, but if the tissue is large, freezing >>>artifact is present i.e. mouse spleen. We do use solid dry ice on occasion >>>to snap freeze extremely tiny NALT, or nasal associated lymphoid tissue >>>from mouse - one of our toughest tissues to dissect out and maintain flat. >>> >>>We never remove the disposable plastic cryomold until just before mounting >>>block on chuck. Obviously your method is very cost effective and you reuse >>>the molds. >>> >>>Thanks for the info, >>> >>>Gayle Callis >>> >>>At 04:17 PM 10/3/2003 -0400, you wrote: >>>>Gayle, >>>> We basically do the same thing you do. We use a stainless steel >square >>>>block sitting in the liquid nitrogen. We fill the foam container with >>>>liquid nitrogen covering the s.s.block, this is left for 5-10 minutes with >>>>adding liquid nitrogen as it goes down. Once the s.s.block is cold enough, >>>>the level of liquid nitrogen is kept below the top of the s.s. block. You >>>>can then set your embedding boat on it. We use the Tissue Tek metal >>>>embedding boats and embedding rings with Shandon's M-1. The metal boat >>>>chills and the M-1 freezes fast enough to keep away the freezing artifact. >>>>The s.s. block is stationary and is easy to set the boat onto. All the >>>>surfaces of the block are flat for cutting. It is easy to do and gives >great >>>>blocks and tissue. While we are collecting tissue, we pop the block out of >>>>the boat and put the block into a plastic bag in another foam container >with >>>>dry ice to keep them frozen until we are done. The blocks never touch the >>>>dry ice and are transferred to a -55 freezer. >>>> Your way is basically the same. I don't see anything wrong with it. >>>> >>>> Gail Macke,HTL >>>> Shriners Burns Hospital--Cincinnati, Ohio >>>> >>>>-----Original Message----- >>>>From: Gayle Callis [mailto:gcallis@montana.edu] >>>>Sent: Friday, October 03, 2003 10:37 AM >>>>To: George Cole; Histonet@lists.utsouthwestern.edu >>>>Subject: further comments on Re: [Histonet] Gayle callis' method of >>>>freezing nerve >>>> >>>> >>>>Correction, or rather clarification to these comments: >>>> >>>>George, >>>> >>>>I do NOT immerse the bottom of the crymold DIRECTLY into pure liquid >>>>nitrogen so this direct contact starts freezing the OCT/tissue/mold bottom >>>>per your description. The cryomold with OCT embedded tissue is placed in >>>>bottom of the EMPTY platic petri dish that has been precooled by FLOATING >>>>the dish on a layer of liquid nitrogen - basically the petri dish is >>>>canoeing on liquid N2! The plastic dish is not as cold as direct liquid >>>>nitrogen contact, acts as a buffer, but still extremely cold to give >>>>perfect artifact free freezing. Freezing this way does not allow for any >>>>curvature of the plastic cryomold, it stays perfectly flat. We only use >>>>Tissue Tek cryomolds as other molds have plastic that is too thick and >>>>conducts heat away slower than the thinner Tissue Tek plastic. One joy of >>>>this method is once the OCT/tissue in cryomold is sitting in cold petri >>>>dish, you can go to the next block immediatetly allowing for a large volume >>>>of blocks/collection session. We do not have to hold a cryomold while it >>>>freezes. It takes a steady hand to prevent liquid nitrogen from making >>>>contact with OCT over the top of cryomold. >>>> >>>>When the OCT finishes freezing to top of mold with petri dish methods, >>>>there are no indentations, just a nice layer of OCT. The petri dish method >>>>is not as fast a freezing as you describe although it results in artifact >>>>free tissues, and the tidge of slower freezing probably means good >>>>interface of tissue to OCT, we never get gaps unless bubbles are not >>>>removed. >>>> >>>>My experience total immersion of OCT embedded tissues into liquid N2 OR >>>>embedded in a mold results in cracked OCT, and a horrible block to section. >>>>However, you are holding the mold to permit freezing starting at bottom, >>>>and as said before - a steady hand helps. >>>> >>>>Our preferred method of snap freezing is dry ice/isopentane slurry, to do a >>>>ton of blocks at one tissue collection session. Only when we need >>>>perfectly flat faced blocks is the petri dish canoeing on liquid nitrogen >>>>used. >>>> >>>>Gayle Callis >>>>MT,HT,HTL(ASCP) >>>>Research Histopathology Supervisor >>>>Veterinary Molecular Biology >>>>Montana State University - Bozeman >>>>PO Box 173610 >>>>Bozeman MT 59717-3610 > >>Philip Oshel >>Supervisor, AMFSC and BBPIC microscopy facilities >>Department of Animal Sciences >>University of Wisconsin >>1675 Observatory Drive >>Madison, WI 53706 - 1284 >>voice: (608) 263-4162 >>fax: (608) 262-5157 (dept. fax) >> >>_______________________________________________ >>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 AmyIchuan <@t> aol.com Mon Oct 6 14:22:56 2003 From: AmyIchuan <@t> aol.com (AmyIchuan@aol.com) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] unsubsrcibe Message-ID: <1d8.11f97444.2cb31b10@aol.com> -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031006/54860340/attachment.htm From Luis.Chiriboga <@t> med.nyu.edu Mon Oct 6 14:42:12 2003 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] COX 1&2 In-Reply-To: Message-ID: have used from Caymen Chemicals catalog #160107 Rabbit anti-human. there is also an antibody available from Transduction labs (Becton Dicknson) catalog #C22420 mouse anti-rat. -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Denise Bland-Piontek Sent: Monday, October 06, 2003 2:00 PM To: histonet@pathology.swmed.edu Subject: [Histonet] COX 1&2 Is anyone currently doing antibodies Cox1 & Cox2? I'm trying to locate a vendor that has a reliable antibody for IHC. So far the only manufacturers I've located don't seem to really have IHC as a priority. As always any help is welcomed! Denise Bland-Piontek WVU _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jim.Tunstead <@t> msnyuhealth.org Mon Oct 6 14:23:44 2003 From: Jim.Tunstead <@t> msnyuhealth.org (Tunstead, Jim) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Histotechnologist position in New York City Message-ID: <1371F1DB5EE3294CA931D4321EBCB004243AD0@EXCEBW2K18.msnyuhealth.org> Dear All, We currently have a position available at Mount Sinai School of Medicine which may be of interest to list users. Please see details below. Histotechnologist / Lab Coordinator We currently have an opening for a Histotechnologist. The individual will be an integral part of a newly equipped pathology core laboratory. The core provides histology, immunohistochemistry and image analysis support for the Cardiovascular Institute. Services are provided to a small group of investigators, allowing greater involvement in their research projects. As well as advising on general procedures, the successful candidate will have the opportunity to develop new processing and staining protocols. Specific Responsibilities: Act as a histology and immunochemistry resource for researchers in the preparation of their protocols. Assist in establishing, optimizing, customizing and documenting protocols for fixing, processing, sectioning, staining, and reagent preparation for research projects. Actively participate in group meetings with researchers to discuss experimental design. Fix, process and paraffin embed samples; process and freeze fresh samples. Perform sectioning of paraffin and frozen blocks for immunohistochemistry, in situ hybridization and special stains. Accurately label, organize, and track tissue, blocks, and slides. Prepare reagents from detailed laboratory procedures and document that quality control standards are being met. Research, troubleshoot, and test new reagents. Inventory, maintain, and order supplies as necessary. Assist in establishing fees and processing billing. Perform routine maintenance on laboratory equipment, including automated immunostainer, microtome, cryostat, freezer, and tissue processor. Assist in organizing work and storage space, and assist in general lab maintenance. Perform other duties, as assigned. The ideal candidate will possess the following qualifications: The minimum requirement is a high school diploma/AA with formal or on the job training as a histology technician. A Bachelor of Science degree and HT/HTL certification by a nationally recognized certifying agency, such as the American Society of Clinical Pathology, are desirable but not required. A minimum of three years of experience is required and good microtome sectioning skills are essential. Additional experience and training are highly desirable, particularly in a research setting. Experience in the use of an automated immunohistology stainer, clinical and research histology experience, and a strong scientific background are very desirable. Experience with microscopy, tissue procurement, and non-human histology is also a plus. The successful technician must be able to organize and prioritize tasks, work accurately and efficiently, be detail oriented, work both independently and with a group, and have excellent written and verbal communication skills. Familiarity with Microsoft Word, Excel, and Internet Explorer is desirable. Business Title: Lab Coordinator Job Type: Full time Job Category: Laboratory research Compensation: Salary DOE + excellent benefits CLOSING DATE: Open until filled. If you have questions or wish to apply, please contact Jim Tunstead by e-mail, phone, fax or mail. Jim Tunstead, Ph.D. Assistant Professor Cardiovascular Institute, Box 1030 Mount Sinai School of Medicine One Gustave Levy Place New York, NY 10029 ph:212-241-0731 (Jacinta Russell) fx: 212-860-7032 jim.tunstead@msnyuhealth.org From gudrun.lang <@t> aon.at Mon Oct 6 14:53:20 2003 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] testicular biopsies Message-ID: <00e901c38c43$7e8aa9e0$0d12a8c0@SERVER> My question is: Is it a must to mix the bouin-solution for fixation of testicular biopsies always freshly? Or is there no difference to "old" solutions? Lang Gudrun Akh Linz, Austria -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031006/234ea7f2/attachment.htm From RITA.ANGEL <@t> UC.EDU Mon Oct 6 15:05:14 2003 From: RITA.ANGEL <@t> UC.EDU (Rita Angel) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] NSH meeting Message-ID: <5.1.0.14.2.20031006160321.032c0508@ucmail3.uc.edu> Thanks to everyone that responded to my question about the NSH meeting this month. Our lab is registered and looking forward to being there. Thanks so much, Rita Angel, HT University of Cincinnati From peggy <@t> nsh.org Mon Oct 6 16:38:55 2003 From: peggy <@t> nsh.org (Peggy Micciche) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] NSH Annual Reports Message-ID: <000001c38c52$3eaabe50$7300000a@histo2> Important Notice To NSH Members: Due to a computer error at the mail-house, your NSH 2003 Annual Report was mailed with an incorrect name. Be assured that our main membership file is correct. If you do not receive a report and you wish to have a copy, please contact the NSH office. We regret any inconvenience this may cause and thank you for your understanding. Sincerely, NSH Office From usdahisto <@t> hotmail.com Mon Oct 6 17:29:22 2003 From: usdahisto <@t> hotmail.com (T. Truscott) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Bird feather Message-ID: Lalen, I would try rinsing your decal solution out of the tissue with water for about ten minutes to make sure all formic acid is gone. then procede as usual. Formic acid left in the tissue can interfere with adhesion. good luck Tom T. USDA Pullman WA. >From: "Sarah Jones" >To: , >Subject: Re: [Histonet] Bird feather >Date: Wed, 01 Oct 2003 23:20:54 -0500 > >Have you tried celloidin coating the sections after deparaffinization? >Are they lifting in the oven before deparaffinization? I have the same >trouble with horse hoof. I use the ultrastick goldseal slides, but >still have trouble from time to time. You could try plastic embedding >as a last resort. Sarah > >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 > > > >>> "Elena Alsing" 10/01/03 04:15PM >>> >Greetings to all! >How do you make bird feather stay on the slide? We received feather >shafts in formalin. I decalcified them in equal amount of sodium citrate >and formic acid and then process them. They cut nicely but they keep on >lifting off the >slide. I put them on plus slides and air dried them over the weekend - >didn't work. I put them on probe-on slides, let them dry dry overnight >at room temp and hand-stained them - didn't work. I don't know what to >do... Please HELP! >Thanks. >Lalen Alsing >UC Davis-CAHFS, Fresno > > > >_______________________________________________ >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 _________________________________________________________________ Instant message in style with MSN Messenger 6.0. Download it now FREE! http://msnmessenger-download.com From Dren6289 <@t> aol.com Mon Oct 6 18:23:36 2003 From: Dren6289 <@t> aol.com (Dren6289@aol.com) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Microwave Tissue PROCESSING Message-ID: <1e7.11109b27.2cb35378@aol.com> Is anyone using a microvwave to do your routine tissue processing. If so, can you tell me the benefits you have found? -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031006/2315c3e1/attachment.htm From Rcartun <@t> harthosp.org Mon Oct 6 18:33:33 2003 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Cyclin D1 Message-ID: The immunohistochemical detection of cyclin D1 in formalin-fixed, paraffin embedded tissue has been problematic. Over the years, I have tried to help many labs detect cyclin D1 by supplying our procedure, as well as positive control tissue. We recently evaluated a new rabbit monoclonal antibody (clone SP4) from Lab Vision Corporation (Fremont, CA) that has proved very impressive in my laboratory (when compared to clone DCS-6). If you are having trouble detecting this protein, I recommend that you try this antibody. Richard Cartun Director, Immunopathology Hartford Hospital Hartford, CT 06102 From micro <@t> superlink.net Mon Oct 6 18:37:37 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Histology Services References: <002401c38c0a$1cf0f1a0$92362850@KEMLOS> Message-ID: <018101c38c62$e3bd8d40$80cd5cd1@DJ4VDH31> Sorry for the many repeated messages! I sent the one reply only, but some smart @$$ keeps repeating my message! Would this person please identify and contact me direct, instead to pollute the list server! ----- Original Message ----- From: "Kemlo Rogerson" To: Sent: Monday, October 06, 2003 9:01 AM Subject: RE: [Histonet] Histology Services So nice you named it twice, um thrice um....... Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 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-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Markus F. Meyenhofer Sent: 03 October 2003 21:22 To: LaTricia Faison; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histology Services Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ 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 micro <@t> superlink.net Mon Oct 6 18:37:37 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Histology Services References: <002401c38c0a$1cf0f1a0$92362850@KEMLOS> Message-ID: <018101c38c62$e3bd8d40$80cd5cd1@DJ4VDH31> Sorry for the many repeated messages! I sent the one reply only, but some smart @$$ keeps repeating my message! Would this person please identify and contact me direct, instead to pollute the list server! ----- Original Message ----- From: "Kemlo Rogerson" To: Sent: Monday, October 06, 2003 9:01 AM Subject: RE: [Histonet] Histology Services So nice you named it twice, um thrice um....... Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 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-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Markus F. Meyenhofer Sent: 03 October 2003 21:22 To: LaTricia Faison; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histology Services Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ 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 micro <@t> superlink.net Mon Oct 6 18:37:37 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Histology Services References: <002401c38c0a$1cf0f1a0$92362850@KEMLOS> Message-ID: <018101c38c62$e3bd8d40$80cd5cd1@DJ4VDH31> Sorry for the many repeated messages! I sent the one reply only, but some smart @$$ keeps repeating my message! Would this person please identify and contact me direct, instead to pollute the list server! ----- Original Message ----- From: "Kemlo Rogerson" To: Sent: Monday, October 06, 2003 9:01 AM Subject: RE: [Histonet] Histology Services So nice you named it twice, um thrice um....... Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 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-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Markus F. Meyenhofer Sent: 03 October 2003 21:22 To: LaTricia Faison; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histology Services Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ 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 micro <@t> superlink.net Mon Oct 6 18:37:37 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Histology Services References: <002401c38c0a$1cf0f1a0$92362850@KEMLOS> Message-ID: <018101c38c62$e3bd8d40$80cd5cd1@DJ4VDH31> Sorry for the many repeated messages! I sent the one reply only, but some smart @$$ keeps repeating my message! Would this person please identify and contact me direct, instead to pollute the list server! ----- Original Message ----- From: "Kemlo Rogerson" To: Sent: Monday, October 06, 2003 9:01 AM Subject: RE: [Histonet] Histology Services So nice you named it twice, um thrice um....... Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 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-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Markus F. Meyenhofer Sent: 03 October 2003 21:22 To: LaTricia Faison; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histology Services Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ 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 adams329 <@t> bellsouth.net Mon Oct 6 19:50:39 2003 From: adams329 <@t> bellsouth.net (adams329@bellsouth.net) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Histology Position in Georgia Message-ID: <20031007005038.ZFTL1821.imf17aec.mail.bellsouth.net@mail.bellsouth.net> We currently have a position for a Histology Technician in Macon, Georgia. The Hospital is HCA Coliseum Medical Center. The hours are 8:30-5:00. Macon is approximately 70 miles South of Atlanta, GA and a 3hour drive to Savannah, GA. The Histology Lab averages about 150 blocks, per day. It is a nice working environment with three certified Histology Techs and one Grossing room Tech. Knowledge of Cytology prep work is a plus. Our fourth Tech is Retiring the end of October. If you're interested, you may email me or contact Radonna Prince-Bush at 478-765-4865. Thank you, Fran Lee Adams,HT(ASCP) Coliseum Medical Center 350 Hospital Drive Macon, GA 31201 From micro <@t> superlink.net Mon Oct 6 18:37:37 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Histology Services References: <002401c38c0a$1cf0f1a0$92362850@KEMLOS> Message-ID: <018101c38c62$e3bd8d40$80cd5cd1@DJ4VDH31> Sorry for the many repeated messages! I sent the one reply only, but some smart @$$ keeps repeating my message! Would this person please identify and contact me direct, instead to pollute the list server! ----- Original Message ----- From: "Kemlo Rogerson" To: Sent: Monday, October 06, 2003 9:01 AM Subject: RE: [Histonet] Histology Services So nice you named it twice, um thrice um....... Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 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-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Markus F. Meyenhofer Sent: 03 October 2003 21:22 To: LaTricia Faison; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histology Services Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ 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 micro <@t> superlink.net Mon Oct 6 18:37:37 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Histology Services References: <002401c38c0a$1cf0f1a0$92362850@KEMLOS> Message-ID: <018101c38c62$e3bd8d40$80cd5cd1@DJ4VDH31> Sorry for the many repeated messages! I sent the one reply only, but some smart @$$ keeps repeating my message! Would this person please identify and contact me direct, instead to pollute the list server! ----- Original Message ----- From: "Kemlo Rogerson" To: Sent: Monday, October 06, 2003 9:01 AM Subject: RE: [Histonet] Histology Services So nice you named it twice, um thrice um....... Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 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-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Markus F. Meyenhofer Sent: 03 October 2003 21:22 To: LaTricia Faison; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histology Services Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ 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 gale-kleven <@t> uiowa.edu Mon Oct 6 21:28:27 2003 From: gale-kleven <@t> uiowa.edu (Gale A. Kleven) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] unsubscribe Message-ID: <001c01c38c7a$b17e5090$a4570640@Hydemobil> ______________________________________________________ Gale A. Kleven Department of Psychology University of Iowa http://www.psychology.uiowa.edu/ E11 Seashore Hall Iowa City, IA 52246 email: gale-kleven@uiowa.edu ______________________________________________________ -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031006/3cdff96b/attachment.htm From FARFAT <@t> aol.com Mon Oct 6 21:33:45 2003 From: FARFAT <@t> aol.com (FARFAT@aol.com) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Cyclin D1 Message-ID: <4a.2315a8b8.2cb38009@aol.com> Hi Richard, I could not agree with you more.CyclinD1 is working great since i got the lab vision product which is rabbit mono.Have you tried their CD-3? it is excellent too. Akbar. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031006/0fc03cf6/attachment.htm From micro <@t> superlink.net Mon Oct 6 18:37:37 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Histology Services References: <002401c38c0a$1cf0f1a0$92362850@KEMLOS> Message-ID: <018101c38c62$e3bd8d40$80cd5cd1@DJ4VDH31> Sorry for the many repeated messages! I sent the one reply only, but some smart @$$ keeps repeating my message! Would this person please identify and contact me direct, instead to pollute the list server! ----- Original Message ----- From: "Kemlo Rogerson" To: Sent: Monday, October 06, 2003 9:01 AM Subject: RE: [Histonet] Histology Services So nice you named it twice, um thrice um....... Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 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-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Markus F. Meyenhofer Sent: 03 October 2003 21:22 To: LaTricia Faison; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histology Services Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ 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 micro <@t> superlink.net Mon Oct 6 18:37:37 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Histology Services References: <002401c38c0a$1cf0f1a0$92362850@KEMLOS> Message-ID: <018101c38c62$e3bd8d40$80cd5cd1@DJ4VDH31> Sorry for the many repeated messages! I sent the one reply only, but some smart @$$ keeps repeating my message! Would this person please identify and contact me direct, instead to pollute the list server! ----- Original Message ----- From: "Kemlo Rogerson" To: Sent: Monday, October 06, 2003 9:01 AM Subject: RE: [Histonet] Histology Services So nice you named it twice, um thrice um....... Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 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-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Markus F. Meyenhofer Sent: 03 October 2003 21:22 To: LaTricia Faison; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histology Services Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ 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 micro <@t> superlink.net Mon Oct 6 18:37:37 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Histology Services References: <002401c38c0a$1cf0f1a0$92362850@KEMLOS> Message-ID: <018101c38c62$e3bd8d40$80cd5cd1@DJ4VDH31> Sorry for the many repeated messages! I sent the one reply only, but some smart @$$ keeps repeating my message! Would this person please identify and contact me direct, instead to pollute the list server! ----- Original Message ----- From: "Kemlo Rogerson" To: Sent: Monday, October 06, 2003 9:01 AM Subject: RE: [Histonet] Histology Services So nice you named it twice, um thrice um....... Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 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-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Markus F. Meyenhofer Sent: 03 October 2003 21:22 To: LaTricia Faison; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histology Services Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ 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 georgecole <@t> ev1.net Tue Oct 7 03:12:47 2003 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] TWO PAGES OF NERVE PROCEDURE Message-ID: <000001c38caa$cc625070$014dbad0@hppav> ALL YOU MUSCLE AND NERVE PACKET RECIPIENTS Two pages containing part of the nerve silver impregnation procedure were left out of the procedure copies mailed to you all. I'm sending those pages to you in a regular manila envelope. Sorry about that---I didn't use the written procedures----and it took me a little while to notice those passages were missing. They were so basic and necessary It just didn't dawn on me that they could turn up missing. I hope that some of you are getting some good out of the packets. georgecole@ev1.net -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031007/5a04a4dd/attachment.htm From micro <@t> superlink.net Mon Oct 6 18:37:37 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Histology Services References: <002401c38c0a$1cf0f1a0$92362850@KEMLOS> Message-ID: <018101c38c62$e3bd8d40$80cd5cd1@DJ4VDH31> Sorry for the many repeated messages! I sent the one reply only, but some smart @$$ keeps repeating my message! Would this person please identify and contact me direct, instead to pollute the list server! ----- Original Message ----- From: "Kemlo Rogerson" To: Sent: Monday, October 06, 2003 9:01 AM Subject: RE: [Histonet] Histology Services So nice you named it twice, um thrice um....... Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 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-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Markus F. Meyenhofer Sent: 03 October 2003 21:22 To: LaTricia Faison; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histology Services Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ 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 micro <@t> superlink.net Mon Oct 6 18:37:37 2003 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Histology Services References: <002401c38c0a$1cf0f1a0$92362850@KEMLOS> Message-ID: <018101c38c62$e3bd8d40$80cd5cd1@DJ4VDH31> Sorry for the many repeated messages! I sent the one reply only, but some smart @$$ keeps repeating my message! Would this person please identify and contact me direct, instead to pollute the list server! ----- Original Message ----- From: "Kemlo Rogerson" To: Sent: Monday, October 06, 2003 9:01 AM Subject: RE: [Histonet] Histology Services So nice you named it twice, um thrice um....... Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 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-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Markus F. Meyenhofer Sent: 03 October 2003 21:22 To: LaTricia Faison; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Histology Services Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ----- Original Message ----- From: "LaTricia Faison" To: Sent: Friday, October 03, 2003 2:18 PM Subject: [Histonet] Histology Services Hello to everyone out there. My name is Trish Faison. I am the manager of a Histology Core Lab at the Medical College of Georgia. We offer a wide range of histology services to fit different research needs. We offer several protocols at a reasonable price with quick turnaround times. If anyone wants more information on the services offered, you can e-mail me or call me at (706)733-0188 ext.2378. _______________________________________________ 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 hodges420 <@t> msn.com Tue Oct 7 02:47:38 2003 From: hodges420 <@t> msn.com (MARY T HODGES) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Position in Tucson, Az Message-ID: Good Morning all, We have need of two histologist here in Tucson in Dr Jill Cohen's Dermpath lab. Anyone wishing to apply fax your resume to Dr Jill Cohen 1-520-320-7684 Thanks Tere Hodges -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031007/c54a6a6a/attachment.htm From c.m.vanderloos <@t> amc.uva.nl Tue Oct 7 06:35:10 2003 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] RE: Cell Culture Staining Message-ID: <8c0e398bbd1f.8bbd1f8c0e39@amc.uva.nl> Hi Kevin, There is a method described by Harold Kerstens to embed your cells into paraffin blocks. Spin the cells down in an Eppendorf tube, remove the supernatant and mix the cells gently with 1 ml warm (65C) 2% agarose solution. Centrifuge again to concentrate the cells. Embed in paraffin as usally. See paper by: Kerstens et al. JHC 48:709-718, 2000. For the next time try to get some collagen sponges from some cell culturing company. Spin the (unfixed!) cells down and wash 3x with PBS (remove proteins). Discard supernatant and resuspend cells in a small volume. Insert sponge and allow the cells to adhere to the collagen (15 min, RT). Fix sponge in buffered formalin and embed as usual. Good luck! Chris van der Loos Dept. of Cardiovascular Pathology Academical Medical Center Amsterdam - The Netherlands >-----Original Message----- >From: Johnson, Kevin [mailto:KJohnson@med.miami.edu] >Sent: Friday, October 03, 2003 3:29 PM >To: 'histonet@lists.utsouthwestern.edu' >Subject: [Histonet] Cell Culture Staining > >Greetings, all. > >A researcher just brought me (at Miller Time on a Friday!) two culture >flasks of terminally differentiated Sertoli cells and requested H&E >staining. Well. >Conventional H&E obviously is out, since polystyrene is not compatible >with xylene. Further restrictions: the cells cannot be scraped off, >they cannot be replated on glass coverslips, and for reasons of her >own, they cannot go back into the incubator pending an answer. They >now have been fixed in situ with 10% neutral buffered formalin. > >Does anyone have a protocol for staining these puppies? If not H&E >per se, then is there another staining method that might satisfy her? > >Thanks in advance, > >Kevin Johnson >University of Miami >Diabetes Research Institute From funderwood <@t> mcohio.org Tue Oct 7 07:42:04 2003 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Purchasing machines Grossing Work Station,Microtom etc. Message-ID: I would highly recommend the tissue tek DRS 2000 automated stainer and to complement that the SCA coverslipper. Bothe are sold by Sakura Finetek. I'm not sure of their web address but call 1.800.725.8723 to get more product info. Fred >>> "Histology" 10/01/03 11:23AM >>> Our lab is in the process of purchasing our automated stainer for routine H&E's,Grossing Work Station,Microtom,Cryotom,Embedding center and processor (300 cassettes). I would really appreciate comments from anyone who really likes, or dislikes the machines that they are using. Our plan is to get an automatic coverslipper as well. Thanks in advance!!! Muhammad Tahseen Histology Supervisor Deptt. Pathology Shaukat Khanum Cancer Hospital And Research Center Lahore. Pakistan Ph. +92 42 5180725-36 Ext 2369, Fax. +92 42 5180723 e-mail. Histology@skm.org.pk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Tue Oct 7 07:42:04 2003 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Purchasing machines Grossing Work Station,Microtom etc. Message-ID: I would highly recommend the tissue tek DRS 2000 automated stainer and to complement that the SCA coverslipper. Bothe are sold by Sakura Finetek. I'm not sure of their web address but call 1.800.725.8723 to get more product info. Fred >>> "Histology" 10/01/03 11:23AM >>> Our lab is in the process of purchasing our automated stainer for routine H&E's,Grossing Work Station,Microtom,Cryotom,Embedding center and processor (300 cassettes). I would really appreciate comments from anyone who really likes, or dislikes the machines that they are using. Our plan is to get an automatic coverslipper as well. Thanks in advance!!! Muhammad Tahseen Histology Supervisor Deptt. Pathology Shaukat Khanum Cancer Hospital And Research Center Lahore. Pakistan Ph. +92 42 5180725-36 Ext 2369, Fax. +92 42 5180723 e-mail. Histology@skm.org.pk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lfaison <@t> mail.mcg.edu Tue Oct 7 08:47:48 2003 From: lfaison <@t> mail.mcg.edu (LaTricia Faison) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Nerve Tissue Message-ID: Does anybody know of a way to unthaw nerve that has been snap frozen to a cork without doing damage to the nerve cells? It's hard to cross section and remove the nerve from the cork when it is attached to it by snap freezing. Thanks in advance for any advice. From DDittus787 <@t> aol.com Tue Oct 7 09:25:11 2003 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] COX 1&2 Message-ID: <09EDAECB.4F9FAB90.0A1F969F@aol.com> you can try zymed as well, recently used both and they worked well. 1-800-874-4494 dana From kalschev <@t> svm.vetmed.wisc.edu Tue Oct 7 11:13:19 2003 From: kalschev <@t> svm.vetmed.wisc.edu (Vicki Kalscheur) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Hard Tissue Committee Meeting Message-ID: <00a901c38ced$ec1a4bf0$65d56880@kalscheur426> Dear members and interested individuals, Please join us on Saturday, October 18th, 2003 from 5:30 - 6:30 pm in Louisville, KY as we gather at NSH's 29th Annual Symposium/Convention. The room number will be posted at the Hard Tissue Display Table. I look forward to seeing many of you! Sincerely, Vicki Kalscheur, Chairperson -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031007/f8fbb953/attachment.htm From MGomez <@t> ameripath.com Tue Oct 7 11:11:54 2003 From: MGomez <@t> ameripath.com (MGomez@ameripath.com) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Histotech needed Message-ID: Good morning Histonetters: We are looking for a certified HT/HTL to work at our Histology Laboratory located in Sandy, Utah. If you know anyone interested please write or call at 801-256-0040 x205/x244 for immediate consideration. You may ask for Milton or Mike. We're located 20 minutes from Salt Lake City and 40 minutes from Park City. Thank you very much, Milton From georgecole <@t> ev1.net Tue Oct 7 11:11:01 2003 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] For the Guiness Book Of Records Message-ID: <000e01c38ced$9adda2a0$074dbad0@hppav> We have a place in the Guiness Book Of Records----Egad!! Who would ever think there would be so many histotechs named Markus F. Meyenhofer. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031007/8d4dc572/attachment.htm From dobbin <@t> upei.ca Tue Oct 7 11:59:51 2003 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] culture cells In-Reply-To: <20031004183831.CF1295BD2E@perceval.uca.es> Message-ID: <3F82C6D8.12995.101E7A6@localhost> Hi Jose, I may be taking this vein even further from the original question, but just a handy little tip for your method. Once the drop of cells has been left on the glass slide for no more than a minute, use a suction device to remove the excess saline (carefully from the side of the drop). I always found that leaving a whole drop to dry, not only took longer, but left an incredible amount of salt crystals that obliterated the preparation. The cells "stick" to the slides fairly quickly due to electrostatic charges and you lose very few cells. The reward is a nice clean, unobscurred preparation. Cheers! Greg Date sent: Sat, 04 Oct 2003 20:38:31 +0200 (CEST) From: Jose Luis Palazon Fernandez Subject: [Histonet] culture cells To: histonet@lists.utsouthwestern.edu Organization: Universidad de Cadiz > maybe you can use trypsine to detach the cells, wash the trypsine out with serum or culture media and them concentrate the cells by centrifugating. After you obtain a concentrated suspention you can drop the cells on the slides using a Pasteur pipete, let them dry and fix with formalin or alcohol, and then treat as a conventional slide for H&E. > > hope this help > > Jos? Luis > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From georgecole <@t> ev1.net Tue Oct 7 13:19:46 2003 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] ROSS STAPF ADDRESS Message-ID: <000001c38cff$9781c840$094dbad0@hppav> Ross, I never did get your complete mailing address. I muscle and nerve packet for you---it's all packed up with no place to go. georgecole@ev1.net -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031007/47078efc/attachment.htm From pruegg <@t> msn.com Tue Oct 7 13:46:14 2003 From: pruegg <@t> msn.com (Patsy Ruegg) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] IHCRG committee meeting Message-ID: I would like to invite those of you attending the NSH S/C to join the IHC Resource Group at our committee meeting Saturday Oct. 18th at 5:30 PM. Check the schedule and the IHCRG committee table for the meeting room number. Patsy Ruegg, IHCRG Committee Chair Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 HP-303-644-4538 HF-303-644-3377 hm email pruegg@msn.com wk email pruegg@colobio.com 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. ----Original Message Follows---- From: "Vicki Kalscheur" To: "Histonet Discussion" Subject: [Histonet] Hard Tissue Committee Meeting Date: Tue, 7 Oct 2003 11:13:19 -0500 Dear members and interested individuals, Please join us on Saturday, October 18th, 2003 from 5:30 - 6:30 pm in Louisville, KY as we gather at NSH's 29th Annual Symposium/Convention. The room number will be posted at the Hard Tissue Display Table. I look forward to seeing many of you! Sincerely, Vicki Kalscheur, Chairperson _________________________________________________________________ Add MSN 8 Internet Software to your existing Internet access and enjoy patented spam protection and more. Sign up now! http://join.msn.com/?page=dept/byoa From kellym <@t> mcmaster.ca Tue Oct 7 14:15:35 2003 From: kellym <@t> mcmaster.ca (Margaret Kelly) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] unsubsrcibe Message-ID: <007901c38d07$65e06c80$1a02a8c0@DELL> -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031007/8f7a7b69/attachment.htm From info <@t> instrumedics.com Tue Oct 7 14:49:26 2003 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] optimizing snap freezing Message-ID: <007801c38d0c$1d2e3f80$6401a8c0@instrumedics1> According to experts in the field of freezing there are two requirements that have to met to minimize the growth of damaging ice crystals. One is rapidly freezing at low temperatures and the other is thermal exchange. The Gentle Jane method embodies both of these requirements. A 3/4lb.chrome-plated brass heat extractor, a good conductor, is chilled in liquid nitrogen (-196C). The heat extractor is placed on the Gentle Jane device and falls at a controlled rate. The tissue to be frozen is placed on the top of the mound of CryoGel or other embedding medium so that the heat extractor contacts the tissue first for the best freezing. The tissue and the embedding medium are frozen simultaneously in 10-15 sec. Please see the details on our web site www.instrumedics.com and go to "products" for the "Stand -Alone Gentle Jane". Bernice schiller@instrumedics.com From Vivian.King <@t> CLS.ab.ca Tue Oct 7 15:10:33 2003 From: Vivian.King <@t> CLS.ab.ca (King, Vivian) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Musto Movat Stain Message-ID: <30C050525B881C4AAFF41E6D16543E68192718@mail3.cls.ab.ca> Does anyone out there have any ideas why the connective tissue is staining up "green" instead of yellow (saffron)? We have been struggling with this for a while and have had no luck in resolving the problem. Any help would be greatly appreciated. Thanks. Vivian King Tech II - Neuropathology FMC Calgary, Alberta 403-944-1576 vivian.king@cls.ab.ca -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031007/b432a6f6/attachment.htm From SJones <@t> cvm.tamu.edu Tue Oct 7 15:41:59 2003 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Musto Movat Stain Message-ID: Hi Vivian, I know that the saffron has to be free of any water contamination. Also, possibly your phosphotungstic acid is old and needs to be changed. 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 >>> "King, Vivian" 10/07/03 03:10PM >>> Does anyone out there have any ideas why the connective tissue is staining up "green" instead of yellow (saffron)? We have been struggling with this for a while and have had no luck in resolving the problem. Any help would be greatly appreciated. Thanks. Vivian King Tech II - Neuropathology FMC Calgary, Alberta 403-944-1576 vivian.king@cls.ab.ca From tigersnake <@t> ecybermind.net Tue Oct 7 19:44:50 2003 From: tigersnake <@t> ecybermind.net (Paul Howard Lockwood) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Histology Services Message-ID: <200310072141.h97LfbW16501@ginsberg.ecybermind.net> To All, I think I see a possible source for this problem of repeating messages. It might not be a person, but a repeater program in the servers some of you use. It might be a bit buggy, in that it sends a message, and then sends it several more times because it hasn't recognized that it has already sent the message. If you are experiencing this problem, call your internet provider and inform them of the problem. They should be able to correct it. I hope this is of some help. Sincerely, Paul Lockwood 10/6/03 4:37:37 PM, "Markus F. Meyenhofer" wrote: >Sorry for the many repeated messages! >I sent the one reply only, but some smart @$$ keeps repeating my message! >Would this person please identify and contact me direct, instead to pollute >the list server! > > >----- Original Message ----- >From: "Kemlo Rogerson" >To: >Sent: Monday, October 06, 2003 9:01 AM >Subject: RE: [Histonet] Histology Services > > >So nice you named it twice, um thrice um....... > >Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS >Tel: 01270 877625 >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-admin@lists.utsouthwestern.edu >[mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Markus F. >Meyenhofer >Sent: 03 October 2003 21:22 >To: LaTricia Faison; histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Histology Services > >Nice ad!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! > > >----- Original Message ----- >From: "LaTricia Faison" >To: >Sent: Friday, October 03, 2003 2:18 PM >Subject: [Histonet] Histology Services > > >Hello to everyone out there. My name is Trish Faison. I am the manager >of >a Histology Core Lab at the Medical College of Georgia. We offer a wide >range of histology services to fit different research needs. We offer >several protocols at a reasonable price with quick turnaround times. If >anyone wants more information on the services offered, you can e-mail me >or >call me at (706)733-0188 ext.2378. > > > > > > > >_______________________________________________ >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 rcthree <@t> u.washington.edu Tue Oct 7 16:59:12 2003 From: rcthree <@t> u.washington.edu (Rob Crawford) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] Lab histo position opening in Seattle Message-ID: <00c901c38d1e$3dd8a790$ecd35f80@RHODOLITE> Hi Everyone- I just wanted to send out notice of a soon to open position in the Seattle area (University of Washington to be specific). My boss will be looking to fill my position in the lab. Ideally, he's looking for someone w/ small animal experience and good cryo histo technique. Immunohistochemistry/Immunofluorescence, alternate embedding techniques(plastic/paraffin), microscopy and digital imaging a plus. No numbers have been mentioned as far as salaries go, but if anyone is interested I'd be happy to talk to them about the lab and/or look at their CV/resume. We're working on putting together an official posting but I wanted to put a notice on Histonet first. If interested it's best to contact me at this email addy. Thanks! Rob Crawford rcthree@u.washington.edu Univ. of Washington Chamberlain Lab 206-221-6386(phone) 206-616-8272(fax) -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031007/535ae908/attachment.htm From IBirchall <@t> groupwise.swin.edu.au Tue Oct 7 18:54:43 2003 From: IBirchall <@t> groupwise.swin.edu.au (Ian Birchall) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] cell culture staining Message-ID: Hi, after fixation with NBF I have performed H+E staining on the cells still adherent to the culture plate. Following fixation, wash with water, cover with haematoxylin (Harris) 5min, wash, blue (if necessary) wash, cover with eosin (3 min), wash and coverslip (on the culture plate) using aqueous mountant (Gurr, aquahere). You can also do immuno staining for cell surface markers on cultured cells without removing them from the plate. Regards, Ian Birchall, Melbourne. >>> histonet-request@lists.utsouthwestern.edu 10/8/03 3:00:01 AM >>> 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-admin@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: Cell Culture Staining (C.M. van der Loos) 2. Re: Purchasing machines Grossing Work Station,Microtom etc. (Fred Underwood) 3. Re: Purchasing machines Grossing Work Station,Microtom etc. (Fred Underwood) 4. Nerve Tissue (LaTricia Faison) 5. Re: COX 1&2 (DDittus787@aol.com) 6. Hard Tissue Committee Meeting (Vicki Kalscheur) 7. Histotech needed (MGomez@ameripath.com) 8. For the Guiness Book Of Records (George Cole) --__--__-- Message: 1 Date: Tue, 07 Oct 2003 13:35:10 +0200 From: "C.M. van der Loos" To: histonet@lists.utsouthwestern.edu Cc: KJohnson@med.miami.edu Subject: [Histonet] RE: Cell Culture Staining Hi Kevin, There is a method described by Harold Kerstens to embed your cells into paraffin blocks. Spin the cells down in an Eppendorf tube, remove the supernatant and mix the cells gently with 1 ml warm (65C) 2% agarose solution. Centrifuge again to concentrate the cells. Embed in paraffin as usally. See paper by: Kerstens et al. JHC 48:709-718, 2000. For the next time try to get some collagen sponges from some cell culturing company. Spin the (unfixed!) cells down and wash 3x with PBS (remove proteins). Discard supernatant and resuspend cells in a small volume. Insert sponge and allow the cells to adhere to the collagen (15 min, RT). Fix sponge in buffered formalin and embed as usual. Good luck! Chris van der Loos Dept. of Cardiovascular Pathology Academical Medical Center Amsterdam - The Netherlands >-----Original Message----- >From: Johnson, Kevin [mailto:KJohnson@med.miami.edu] >Sent: Friday, October 03, 2003 3:29 PM >To: 'histonet@lists.utsouthwestern.edu' >Subject: [Histonet] Cell Culture Staining > >Greetings, all. > >A researcher just brought me (at Miller Time on a Friday!) two culture >flasks of terminally differentiated Sertoli cells and requested H&E >staining. Well. >Conventional H&E obviously is out, since polystyrene is not compatible >with xylene. Further restrictions: the cells cannot be scraped off, >they cannot be replated on glass coverslips, and for reasons of her >own, they cannot go back into the incubator pending an answer. They >now have been fixed in situ with 10% neutral buffered formalin. > >Does anyone have a protocol for staining these puppies? If not H&E >per se, then is there another staining method that might satisfy her? > >Thanks in advance, > >Kevin Johnson >University of Miami >Diabetes Research Institute --__--__-- Message: 2 Date: Tue, 07 Oct 2003 08:42:04 -0400 From: "Fred Underwood" To: , Subject: Re: [Histonet] Purchasing machines Grossing Work Station,Microtom etc. I would highly recommend the tissue tek DRS 2000 automated stainer and to complement that the SCA coverslipper. Bothe are sold by Sakura Finetek. I'm not sure of their web address but call 1.800.725.8723 to get more product info. Fred >>> "Histology" 10/01/03 11:23AM >>> Our lab is in the process of purchasing our automated stainer for routine H&E's,Grossing Work Station,Microtom,Cryotom,Embedding center and processor (300 cassettes). I would really appreciate comments from anyone who really likes, or dislikes the machines that they are using. Our plan is to get an automatic coverslipper as well. Thanks in advance!!! Muhammad Tahseen Histology Supervisor Deptt. Pathology Shaukat Khanum Cancer Hospital And Research Center Lahore. Pakistan Ph. +92 42 5180725-36 Ext 2369, Fax. +92 42 5180723 e-mail. Histology@skm.org.pk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --__--__-- Message: 3 Date: Tue, 07 Oct 2003 08:42:04 -0400 From: "Fred Underwood" To: , Subject: Re: [Histonet] Purchasing machines Grossing Work Station,Microtom etc. I would highly recommend the tissue tek DRS 2000 automated stainer and to complement that the SCA coverslipper. Bothe are sold by Sakura Finetek. I'm not sure of their web address but call 1.800.725.8723 to get more product info. Fred >>> "Histology" 10/01/03 11:23AM >>> Our lab is in the process of purchasing our automated stainer for routine H&E's,Grossing Work Station,Microtom,Cryotom,Embedding center and processor (300 cassettes). I would really appreciate comments from anyone who really likes, or dislikes the machines that they are using. Our plan is to get an automatic coverslipper as well. Thanks in advance!!! Muhammad Tahseen Histology Supervisor Deptt. Pathology Shaukat Khanum Cancer Hospital And Research Center Lahore. Pakistan Ph. +92 42 5180725-36 Ext 2369, Fax. +92 42 5180723 e-mail. Histology@skm.org.pk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --__--__-- Message: 4 Date: Tue, 07 Oct 2003 09:47:48 -0400 From: "LaTricia Faison" To: Subject: [Histonet] Nerve Tissue Does anybody know of a way to unthaw nerve that has been snap frozen to a = cork without doing damage to the nerve cells? It's hard to cross section = and remove the nerve from the cork when it is attached to it by snap = freezing. Thanks in advance for any advice. --__--__-- Message: 5 Date: Tue, 07 Oct 2003 10:25:11 -0400 From: DDittus787@aol.com To: Luis.Chiriboga@med.nyu.edu, dbpiontek@hsc.wvu.edu, histonet@pathology.swmed.edu Subject: Re: [Histonet] COX 1&2 you can try zymed as well, recently used both and they worked well. 1-800-874-4494 dana --__--__-- Message: 6 From: "Vicki Kalscheur" To: "Histonet Discussion" Date: Tue, 7 Oct 2003 11:13:19 -0500 Subject: [Histonet] Hard Tissue Committee Meeting This is a multi-part message in MIME format. ------=_NextPart_000_00A6_01C38CC4.03368850 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable Dear members and interested individuals, Please join us on Saturday, October 18th, 2003 from 5:30 - 6:30 pm = in Louisville, KY as we gather at NSH's 29th Annual = Symposium/Convention. The room number will be posted at the Hard Tissue = Display Table. I look forward to seeing many of you!=20 Sincerely,=20 Vicki Kalscheur, Chairperson ------=_NextPart_000_00A6_01C38CC4.03368850 Content-Type: text/html; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable
Dear members and interested=20 individuals,
 
    Please join us on = Saturday,=20 October 18th, 2003 from 5:30 - 6:30 pm in Louisville, KY as we gather at = NSH's=20 29th Annual Symposium/Convention.  The room number will be posted = at the=20 Hard Tissue Display Table. I look forward to seeing many of you! =
 
Sincerely, 
    Vicki Kalscheur,=20 Chairperson
------=_NextPart_000_00A6_01C38CC4.03368850-- --__--__-- Message: 7 From: MGomez@ameripath.com To: Histonet@lists.utsouthwestern.edu Date: Tue, 7 Oct 2003 12:11:54 -0400 Subject: [Histonet] Histotech needed Good morning Histonetters: We are looking for a certified HT/HTL to work at our Histology Laboratory located in Sandy, Utah. If you know anyone interested please write or call at 801-256-0040 x205/x244 for immediate consideration. You may ask for Milton or Mike. We're located 20 minutes from Salt Lake City and 40 minutes from Park City. Thank you very much, Milton --__--__-- Message: 8 From: "George Cole" To: Date: Tue, 7 Oct 2003 09:11:01 -0700 Subject: [Histonet] For the Guiness Book Of Records This is a multi-part message in MIME format. ------=_NextPart_000_000F_01C38CB2.EE7ECAA0 Content-Type: text/plain; charset="us-ascii" Content-Transfer-Encoding: 7bit We have a place in the Guiness Book Of Records----Egad!! Who would ever think there would be so many histotechs named Markus F. Meyenhofer. ------=_NextPart_000_000F_01C38CB2.EE7ECAA0 Content-Type: text/html; charset="us-ascii" Content-Transfer-Encoding: quoted-printable

We have a place in the Guiness Book Of Records----Egad!! Who would ever = think there would be so many histotechs = named

 Markus = F. Meyenhofer.   

------=_NextPart_000_000F_01C38CB2.EE7ECAA0-- --__--__-- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest From kemlo <@t> tiscali.co.uk Wed Oct 8 02:35:50 2003 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:22:00 2005 Subject: [Histonet] For the Guiness Book Of Records In-Reply-To: <000e01c38ced$9adda2a0$074dbad0@hppav> Message-ID: <002301c38d6e$cf8450e0$4d122850@KEMLOS> Methinks he is the victim of a cunning virus. Methinks he is the victim of a cunning virus. Methinks he is the victim of a cunning virus. Methinks he is the victim of a cunning virus. Methinks he is the victim of a cunning virus. Methinks he is the victim of a cunning virus. Methinks he is the victim of a cunning virus. Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 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-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of George Cole Sent: 07 October 2003 17:11 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] For the Guiness Book Of Records We have a place in the Guiness Book Of Records----Egad!! Who would ever think there would be so many histotechs named Markus F. Meyenhofer. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/fd2ff21e/attachment.htm From mwhitesi <@t> adventisthealthcare.com Wed Oct 8 05:38:28 2003 From: mwhitesi <@t> adventisthealthcare.com (Marnie Whiteside) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Position in Maryland Message-ID: We are currently looking for a histology assistant to work in the gross from 830am to 5pm Monday through Friday. There is the opportunity for advancement. We have trained several of our histology assistants to become histology technicians. We are located in the Tacoma Park, Maryland just outside of Washington DC. If you are interested you can reach me at 301-891-6102/5155. Marnie Whiteside Washington Adventist Hospital -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/c4b86568/attachment.htm From frouwke <@t> sci.kun.nl Wed Oct 8 05:47:14 2003 From: frouwke <@t> sci.kun.nl (Frouwke Kuijpers) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] RE: shrinkage and Perfusion One system References: Message-ID: <00e901c38d89$8a53f5a0$4a8aae83@sci.kun.nl> Start the prewash at low pressure, and pump up to 300 mmHg to break the blood brain barrier and wash the extracellular fluid (and red blood cells). Charles: next question: we don't do the perfusion by a pump, just by gravity. You said the pressure for the 10% sucrose must go till mm300Hg.... Well we dont' have a pump: what will your advice be than? Start with the saline solution to get rid of the blood and than the sucrose? Frouwke Kuijpers ----- Original Message ----- From: "Charles W. Scouten, Ph.D." To: ; Sent: Monday, October 06, 2003 3:53 PM Subject: [Histonet] RE: shrinkage and Perfusion One system The saline prewash would be just lost time. It accomplishes nothing that will not be accomplished by the sucrose, which is equally effective at washing out the blood (more effective given the high pressure). Start with the sucrose. Time is of the essence, tissue deterioration begins immediately upon anoxia. Switch to fixative when the animal's muscles cease to move. 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 www.myneurolab.com -----Original Message----- From: Nancy.Walker@sanofi-synthelabo.com [mailto:Nancy.Walker@sanofi-synthelabo.com] Sent: Friday, October 03, 2003 8:12 AM To: Charles W. Scouten, Ph.D.; Histonet@lists.utsouthwestern.edu Subject: shrinkage and Perfusion One system Hello, >From the website mentioned in C. Scouten's mail I recuperated this info: The shrinkage occurs when a prewash with physiological saline is followed by fixative. The first action of fixative on the cell membrane is to shut off the sodium pump proteins. Sodium rushes into the cell, followed by water to maintain tonicity, and cells swell and expand. Membrane proteins are fixed and crosslinked to neighboring cells in the swollen position. Later, the cells stabilize and contract, and pull other cells with them. The result is gross shrinkage, local distortion and torn membranes with lost cellular contents from some cells. Tissue Shrinkage can be completely prevented if the extracellular fluid with ions is replaced by a nonionic isotonic solution that cannot enter the cells. This can be accomplished by prewash with 5% (isotonic) sucrose in distilled water. Start the prewash at low pressure, and pump up to 300 mmHg to break the blood brain barrier and wash the extracellular fluid (and red blood cells). Switch shortly thereafter to fixative. The perfusion takes the same time as the traditional procedure, and accomplishes the favorable results bulleted above. So if I'd like to try out the perfusion techniques that are made simple by the Perfusion one system, what would you suggest: first do a physiological saline solution, followed by a prewash with 5% sucrose and then PFA, or go directly to 5% sucrose then PFA? thanks for your advice, Nancy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mucram11 <@t> earthlink.net Wed Oct 8 08:06:26 2003 From: mucram11 <@t> earthlink.net (mucram11@earthlink.net) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] RE: shrinkage and Perfusion One system Message-ID: <57050-220031038136265@M2W059.mail2web.com> Hi, We never had a pump. We used gravity for over 2,000 rats so I think I can say it will work. The height of the container or IV bottle used seemed to be the key. We controlled flow with the standard IV roller closure on the tubing. It was approximately 3ft above the area the animal was being perfused in, always under a hood or very well ventilated area. We looked for a steady drip in the tubing and out the cannula. We did not open the valve all the way as it would blow out the vascular system. We did have a couple of students try wide open and prove the point very well. The use of a saline solution with and without anticoag agents had been used. If you are going to use sucrose I would use the pre-wash to get the blood and red cells out. Just to be sure you have all the small vessels cleared for the fixative or freezing technique. Pam Marcum Original Message: ----------------- From: Frouwke Kuijpers frouwke@sci.kun.nl Date: Wed, 8 Oct 2003 12:47:14 +0200 To: cwscouten@myneurolab.com, Nancy.Walker@sanofi-synthelabo.com, Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: shrinkage and Perfusion One system Start the prewash at low pressure, and pump up to 300 mmHg to break the blood brain barrier and wash the extracellular fluid (and red blood cells). Charles: next question: we don't do the perfusion by a pump, just by gravity. You said the pressure for the 10% sucrose must go till mm300Hg.... Well we dont' have a pump: what will your advice be than? Start with the saline solution to get rid of the blood and than the sucrose? Frouwke Kuijpers ----- Original Message ----- From: "Charles W. Scouten, Ph.D." To: ; Sent: Monday, October 06, 2003 3:53 PM Subject: [Histonet] RE: shrinkage and Perfusion One system The saline prewash would be just lost time. It accomplishes nothing that will not be accomplished by the sucrose, which is equally effective at washing out the blood (more effective given the high pressure). Start with the sucrose. Time is of the essence, tissue deterioration begins immediately upon anoxia. Switch to fixative when the animal's muscles cease to move. 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 www.myneurolab.com -----Original Message----- From: Nancy.Walker@sanofi-synthelabo.com [mailto:Nancy.Walker@sanofi-synthelabo.com] Sent: Friday, October 03, 2003 8:12 AM To: Charles W. Scouten, Ph.D.; Histonet@lists.utsouthwestern.edu Subject: shrinkage and Perfusion One system Hello, >From the website mentioned in C. Scouten's mail I recuperated this info: The shrinkage occurs when a prewash with physiological saline is followed by fixative. The first action of fixative on the cell membrane is to shut off the sodium pump proteins. Sodium rushes into the cell, followed by water to maintain tonicity, and cells swell and expand. Membrane proteins are fixed and crosslinked to neighboring cells in the swollen position. Later, the cells stabilize and contract, and pull other cells with them. The result is gross shrinkage, local distortion and torn membranes with lost cellular contents from some cells. Tissue Shrinkage can be completely prevented if the extracellular fluid with ions is replaced by a nonionic isotonic solution that cannot enter the cells. This can be accomplished by prewash with 5% (isotonic) sucrose in distilled water. Start the prewash at low pressure, and pump up to 300 mmHg to break the blood brain barrier and wash the extracellular fluid (and red blood cells). Switch shortly thereafter to fixative. The perfusion takes the same time as the traditional procedure, and accomplishes the favorable results bulleted above. So if I'd like to try out the perfusion techniques that are made simple by the Perfusion one system, what would you suggest: first do a physiological saline solution, followed by a prewash with 5% sucrose and then PFA, or go directly to 5% sucrose then PFA? thanks for your advice, Nancy _______________________________________________ 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 -------------------------------------------------------------------- mail2web - Check your email from the web at http://mail2web.com/ . From mbecker <@t> pathlabinc.com Wed Oct 8 09:15:50 2003 From: mbecker <@t> pathlabinc.com (Michelle Becker) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] test Message-ID: From siksik <@t> vgernet.net Wed Oct 8 09:24:34 2003 From: siksik <@t> vgernet.net (Steven Slap) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] freezing in isopentane Message-ID: Hi HistoNetters At risk of someone writing "nice ad" again, there is an instrument, the "Clini-RF", from Bright Instruments, that would be very useful in resolving some of these issues. Instead of using liquid nitrogen to cool the isopentane, the Clini-RF is a stand-alone freezer with a -80 degree tank which can be used for direct immersion in isopentane. Although this instrument is available from Hacker Instruments in the United States, I get nothing except satisfaction from seeing it sold. best regards, Steven Slap From cmconway <@t> usgs.gov Wed Oct 8 08:39:02 2003 From: cmconway <@t> usgs.gov (Carla M Conway) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Azure B melanin stain protocol wanted Message-ID: Does anyone have a protocol for Azure B staining of melanin in FFPE tissues? I will be using Dako's Envision Doublestain system (which comes with DAB and Fast Red chromogens) for IHC of trout kidney and want to avoid melanin bleaching with potassium permangante. Thanks in advance, Carla Conway Western Fisheries Research Center 6505 NE 65th St. Seattle, WA ph: 206-526-6282 ext. 242 fax: 206-526-6654 From STapper <@t> slhduluth.com Wed Oct 8 09:43:52 2003 From: STapper <@t> slhduluth.com (Tapper, Sheila) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Node revealing solution Message-ID: <3BFBBD68413CB443A7125A63EC0ACD1D6716A4@slhw2smail01.slhdomain.com> I am looking for a commercial preparation of the node revealing solution used at the grossing bench. We have been preparing a solution here that consists of acetic acid, buffered formalin, 95% ethanol, and diethyl ether. Is anyone aware of a vendor? Thank you. Sheila Tapper St. Luke's Hospital Duluth, MN -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/f0802944/attachment.htm From Ronnie_Houston <@t> bshsi.com Wed Oct 8 10:07:06 2003 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Node revealing solution Message-ID: <530361BF03351B4CAE5270A05D3037B5FE0479@bsrexms01.BSHSIR.COM> Decal Corporation, 800 428 5856, www.decal-bone.com has a product Dissect-Aid that is designed for that purpose.. Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 ronnie_houston@bshsi.com -----Original Message----- From: Tapper, Sheila [mailto:STapper@slhduluth.com] Sent: Wednesday, October 08, 2003 10:44 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Node revealing solution I am looking for a commercial preparation of the node revealing solution used at the grossing bench. We have been preparing a solution here that consists of acetic acid, buffered formalin, 95% ethanol, and diethyl ether. Is anyone aware of a vendor? Thank you. Sheila Tapper St. Luke's Hospital Duluth, MN ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ 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. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/5c501240/attachment.htm From cfavara <@t> niaid.nih.gov Wed Oct 8 10:23:54 2003 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Node revealing solution Message-ID: I used Pen-fix from Anatech for nodes in rabbits and found it very useful. I think it is about the same thing! Best regards, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: Tapper, Sheila [mailto:STapper@slhduluth.com] Sent: Wednesday, October 08, 2003 8:44 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Node revealing solution I am looking for a commercial preparation of the node revealing solution used at the grossing bench. We have been preparing a solution here that consists of acetic acid, buffered formalin, 95% ethanol, and diethyl ether. Is anyone aware of a vendor? Thank you. Sheila Tapper St. Luke's Hospital Duluth, MN -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/e5371300/attachment.htm From StarkusL <@t> ummhc.org Wed Oct 8 10:29:38 2003 From: StarkusL <@t> ummhc.org (Starkus, Laurie) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Mouse tissue Message-ID: I've just started doing research with mouse tissue and had some difficulty flattening the tissue in the water bath so that I got no folds in the tissue. I've addressed this issue by turning up the temperature on the water bath. However, now, I am getting a complaint that some of the tissues are damaged. Some of the tissue is disattached from the main tissue. What temperature do you all use your water baths at? I'm using gelatin in the water bath and coated slides. Anyone have any advice on cutting mouse tissue. I've never had this problem with human tissue. From Cheryl.Clarke <@t> fraserhealth.ca Wed Oct 8 10:35:30 2003 From: Cheryl.Clarke <@t> fraserhealth.ca (Clarke, Cheryl) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] unsubsrcibe Message-ID: From KMMerril <@t> LancasterGeneral.org Wed Oct 8 10:36:52 2003 From: KMMerril <@t> LancasterGeneral.org (Merrill, Kathleen M) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] biopsy artifact Message-ID: We are experiencing a purple haze(Jimi Hendrix) artifact on primarily GI biopsies. We have played with the times on our short bx processing cycle to no avail. We would like any info on biopsy cycles(times, reagents, etc) and any suggestions to improve and rid ourselves of this problem Thanks. Kathleen Merrill, HT. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/e442b678/attachment.htm From cwscouten <@t> myneurolab.com Wed Oct 8 10:39:33 2003 From: cwscouten <@t> myneurolab.com (Charles W. Scouten, Ph.D.) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] RE: shrinkage and Perfusion One system Message-ID: Yes, gravity will work, with 20% shrinkage of the brain, some shape distortion, and with a reddish tinge due to remaining red blood cells, which catalyze any HRP reaction, and autofluoresce. OK for some purposes, terrible for others, but not the optimum for any. I have asked www.Histonet.org to post a picture called Perfused Brains Annotated which shows whole rat brains, one fresh dissected out, one perfused by pressure sucrose and 4% paraformaldehyde/glut, and the last by traditional gravity flow with saline prewash, and the same fixative. The last is reddish and shrunken compared to the middle brain. Three ft of water translates directly, using the units converter at: http://www.sciencemadesimple.com/conversions.html to 67 mm Hg, below average diastolic pressure in mice or rats or people. And some pressure is lost in the lines and needle. Even in living animals with cycles of systolic pressure, some capillaries occasionally get plugged for a while and stop flowing. That is part of the advantage of exercise, to keep the lines open. It is extremely unlikely that completely open exposure to 67 mm Hg would in any way "blow" the vascular system of any mammal, or even clear out all the blood. The blocked passages will stop fixative from reaching the surrounding tissue in that area, and lead to local deterioration in unpredictable and irreproduceible areas. The only advantage of sucrose over saline is to clear the extracellular fluid of sodium before fixative hits. High pressure, 300 mm Hg, applied to either saline or sucrose as the prewash, (there is never a reason for both), will thoroughly clear all red blood cells, but should not be pumped up in advance the 'thump' the system, but increase over a few seconds to clear most blood before the blood brain barrier opens. This prewash will take half the time of traditional prewash, and get fixative in sooner and more thoroughly. 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 www.myneurolab.com -----Original Message----- From: mucram11@earthlink.net [mailto:mucram11@earthlink.net] Sent: Wednesday, October 08, 2003 8:06 AM To: frouwke@sci.kun.nl; Charles W. Scouten, Ph.D.; nancy.walker@sanofi-synthelabo.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: shrinkage and Perfusion One system Hi, We never had a pump. We used gravity for over 2,000 rats so I think I can say it will work. The height of the container or IV bottle used seemed to be the key. We controlled flow with the standard IV roller closure on the tubing. It was approximately 3ft above the area the animal was being perfused in, always under a hood or very well ventilated area. We looked for a steady drip in the tubing and out the cannula. We did not open the valve all the way as it would blow out the vascular system. We did have a couple of students try wide open and prove the point very well. The use of a saline solution with and without anticoag agents had been used. If you are going to use sucrose I would use the pre-wash to get the blood and red cells out. Just to be sure you have all the small vessels cleared for the fixative or freezing technique. Pam Marcum Original Message: ----------------- From: Frouwke Kuijpers frouwke@sci.kun.nl Date: Wed, 8 Oct 2003 12:47:14 +0200 To: cwscouten@myneurolab.com, Nancy.Walker@sanofi-synthelabo.com, Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: shrinkage and Perfusion One system Start the prewash at low pressure, and pump up to 300 mmHg to break the blood brain barrier and wash the extracellular fluid (and red blood cells). Charles: next question: we don't do the perfusion by a pump, just by gravity. You said the pressure for the 10% sucrose must go till mm300Hg.... Well we dont' have a pump: what will your advice be than? Start with the saline solution to get rid of the blood and than the sucrose? Frouwke Kuijpers ----- Original Message ----- From: "Charles W. Scouten, Ph.D." To: ; Sent: Monday, October 06, 2003 3:53 PM Subject: [Histonet] RE: shrinkage and Perfusion One system The saline prewash would be just lost time. It accomplishes nothing that will not be accomplished by the sucrose, which is equally effective at washing out the blood (more effective given the high pressure). Start with the sucrose. Time is of the essence, tissue deterioration begins immediately upon anoxia. Switch to fixative when the animal's muscles cease to move. 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 www.myneurolab.com -----Original Message----- From: Nancy.Walker@sanofi-synthelabo.com [mailto:Nancy.Walker@sanofi-synthelabo.com] Sent: Friday, October 03, 2003 8:12 AM To: Charles W. Scouten, Ph.D.; Histonet@lists.utsouthwestern.edu Subject: shrinkage and Perfusion One system Hello, >From the website mentioned in C. Scouten's mail I recuperated this info: The shrinkage occurs when a prewash with physiological saline is followed by fixative. The first action of fixative on the cell membrane is to shut off the sodium pump proteins. Sodium rushes into the cell, followed by water to maintain tonicity, and cells swell and expand. Membrane proteins are fixed and crosslinked to neighboring cells in the swollen position. Later, the cells stabilize and contract, and pull other cells with them. The result is gross shrinkage, local distortion and torn membranes with lost cellular contents from some cells. Tissue Shrinkage can be completely prevented if the extracellular fluid with ions is replaced by a nonionic isotonic solution that cannot enter the cells. This can be accomplished by prewash with 5% (isotonic) sucrose in distilled water. Start the prewash at low pressure, and pump up to 300 mmHg to break the blood brain barrier and wash the extracellular fluid (and red blood cells). Switch shortly thereafter to fixative. The perfusion takes the same time as the traditional procedure, and accomplishes the favorable results bulleted above. So if I'd like to try out the perfusion techniques that are made simple by the Perfusion one system, what would you suggest: first do a physiological saline solution, followed by a prewash with 5% sucrose and then PFA, or go directly to 5% sucrose then PFA? thanks for your advice, Nancy _______________________________________________ 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 -------------------------------------------------------------------- mail2web - Check your email from the web at http://mail2web.com/ . From Jackie.O'Connor <@t> abbott.com Wed Oct 8 10:55:44 2003 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] biopsy artifact Message-ID: What type of fixative are you using? I remember Jimi - saw him in concert at the Hollywood Bowl - he was the opening act for the Mama's and Papa's in 1966. Anyway, back from my trip - - -I remember trying a formalin substitute at one hospital where we did a lot of GI biopsies - the pathologists complained about a purple haze! Unfortunately, my Jimi Hendrix days won't let me remember the type of formalin substitute - I'm thinking some kind of zinc - but not sure. Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics 847.938.4919 "Merrill, Kathleen M" Sent by: histonet-admin@lists.utsouthwestern.edu 10/08/2003 10:36 AM To: cc: Subject: [Histonet] biopsy artifact We are experiencing a purple haze(Jimi Hendrix) artifact on primarily GI biopsies. We have played with the times on our short bx processing cycle to no avail. We would like any info on biopsy cycles(times, reagents, etc) and any suggestions to improve and rid ourselves of this problem Thanks. Kathleen Merrill, HT. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/443c152f/attachment.htm From ctsmolde <@t> capeheart.uct.ac.za Wed Oct 8 02:34:39 2003 From: ctsmolde <@t> capeheart.uct.ac.za (Jenny Molde) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Cost Message-ID: Hi everyone out there I would like to know more or less what one would charge for a routine H+E and special stain on a sample recieved from explant and embedded in MMA resin, cut and stained. Also the immunocytochemical charge please. All suggestions will be greatly appreciated. Many thanks. Jenny Molde Cardiovascular Research Unit University of Cape Town South Africa. From livieira <@t> ualg.pt Wed Oct 8 04:49:49 2003 From: livieira <@t> ualg.pt (Lina Vieira) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Xilol and xilol substitute Message-ID: <000b01c38d81$83f169c0$2914100a@labhistologia> Helo Everyone, Can you give me some advise about use xilol substitute in the tissue prossessing and after, xilol in the staining? I have this question because in this laboratory we use only xilol substitute (terpene based), and someone ask me about the possibility of prossessing and cut the tissue in this laboratory, and the staining would be make in another laboratory... Tanks in advance Lina Vieira -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/63e64b4f/attachment.htm From lfaison <@t> mail.mcg.edu Wed Oct 8 11:26:42 2003 From: lfaison <@t> mail.mcg.edu (LaTricia Faison) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Shattering Mouse Brain Message-ID: Does anyone know what may cause mouse brain to separate when put on the water bath? The tissue was given to me after it had been in paraformaldehyde for about 7-8 months. It was paraffin processed and shattered soon after touching the water bath. From Ronnie_Houston <@t> bshsi.com Wed Oct 8 11:33:22 2003 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Association for Molecular Pathology Message-ID: <530361BF03351B4CAE5270A05D3037B5FE047B@bsrexms01.BSHSIR.COM> Anyone going to the annual meeting of the Association of Molecular Pathology meeting in Orlando, Nov 20-23, 2003? Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 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 edmondsj <@t> musc.edu Wed Oct 8 11:42:25 2003 From: edmondsj <@t> musc.edu (edmondsj) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Shattering Mouse Brain In-Reply-To: References: Message-ID: <18360538.1065616945@jbepc.thurmond-gazes.musc.edu> Your water bath may be too high, neuro tissue tends to explode on a water bath that is higher than 38-40 degrees celcius. Try cooling the bath down. Joyce Edmonds MUSC Charleston, SC --On Wednesday, October 08, 2003 12:26 PM -0400 LaTricia Faison wrote: > Does anyone know what may cause mouse brain to separate when put on the > water bath? The tissue was given to me after it had been in > paraformaldehyde for about 7-8 months. It was paraffin processed and > shattered soon after touching the water bath. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Lynne.Bell <@t> hitchcock.org Wed Oct 8 11:40:34 2003 From: Lynne.Bell <@t> hitchcock.org (Bell, Lynne) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Node revealing solution Message-ID: We use a product called "Dissect Aid" manufactured by the Decal Chemical Corporation, phone number is 800-428-5856. It contains water, glacial acetic acid, alcohol and formaldehyde. Our pathologists really like it. Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/7c0fd42e/attachment.htm From mucram11 <@t> earthlink.net Wed Oct 8 12:19:33 2003 From: mucram11 <@t> earthlink.net (mucram11@earthlink.net) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] RE: shrinkage and Perfusion One system Message-ID: <244640-220031038171933309@M2W047.mail2web.com> Strange we did not have shrinkage problems with our set up. Pam Marcum Original Message: ----------------- From: Charles W. Scouten, Ph.D. cwscouten@myneurolab.com Date: Wed, 8 Oct 2003 10:39:33 -0500 To: mucram11@earthlink.net, frouwke@sci.kun.nl, nancy.walker@sanofi-synthelabo.com, histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: shrinkage and Perfusion One system Yes, gravity will work, with 20% shrinkage of the brain, some shape distortion, and with a reddish tinge due to remaining red blood cells, which catalyze any HRP reaction, and autofluoresce. OK for some purposes, terrible for others, but not the optimum for any. I have asked www.Histonet.org to post a picture called Perfused Brains Annotated which shows whole rat brains, one fresh dissected out, one perfused by pressure sucrose and 4% paraformaldehyde/glut, and the last by traditional gravity flow with saline prewash, and the same fixative. The last is reddish and shrunken compared to the middle brain. Three ft of water translates directly, using the units converter at: http://www.sciencemadesimple.com/conversions.html to 67 mm Hg, below average diastolic pressure in mice or rats or people. And some pressure is lost in the lines and needle. Even in living animals with cycles of systolic pressure, some capillaries occasionally get plugged for a while and stop flowing. That is part of the advantage of exercise, to keep the lines open. It is extremely unlikely that completely open exposure to 67 mm Hg would in any way "blow" the vascular system of any mammal, or even clear out all the blood. The blocked passages will stop fixative from reaching the surrounding tissue in that area, and lead to local deterioration in unpredictable and irreproduceible areas. The only advantage of sucrose over saline is to clear the extracellular fluid of sodium before fixative hits. High pressure, 300 mm Hg, applied to either saline or sucrose as the prewash, (there is never a reason for both), will thoroughly clear all red blood cells, but should not be pumped up in advance the 'thump' the system, but increase over a few seconds to clear most blood before the blood brain barrier opens. This prewash will take half the time of traditional prewash, and get fixative in sooner and more thoroughly. 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 www.myneurolab.com -----Original Message----- From: mucram11@earthlink.net [mailto:mucram11@earthlink.net] Sent: Wednesday, October 08, 2003 8:06 AM To: frouwke@sci.kun.nl; Charles W. Scouten, Ph.D.; nancy.walker@sanofi-synthelabo.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: shrinkage and Perfusion One system Hi, We never had a pump. We used gravity for over 2,000 rats so I think I can say it will work. The height of the container or IV bottle used seemed to be the key. We controlled flow with the standard IV roller closure on the tubing. It was approximately 3ft above the area the animal was being perfused in, always under a hood or very well ventilated area. We looked for a steady drip in the tubing and out the cannula. We did not open the valve all the way as it would blow out the vascular system. We did have a couple of students try wide open and prove the point very well. The use of a saline solution with and without anticoag agents had been used. If you are going to use sucrose I would use the pre-wash to get the blood and red cells out. Just to be sure you have all the small vessels cleared for the fixative or freezing technique. Pam Marcum Original Message: ----------------- From: Frouwke Kuijpers frouwke@sci.kun.nl Date: Wed, 8 Oct 2003 12:47:14 +0200 To: cwscouten@myneurolab.com, Nancy.Walker@sanofi-synthelabo.com, Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: shrinkage and Perfusion One system Start the prewash at low pressure, and pump up to 300 mmHg to break the blood brain barrier and wash the extracellular fluid (and red blood cells). Charles: next question: we don't do the perfusion by a pump, just by gravity. You said the pressure for the 10% sucrose must go till mm300Hg.... Well we dont' have a pump: what will your advice be than? Start with the saline solution to get rid of the blood and than the sucrose? Frouwke Kuijpers ----- Original Message ----- From: "Charles W. Scouten, Ph.D." To: ; Sent: Monday, October 06, 2003 3:53 PM Subject: [Histonet] RE: shrinkage and Perfusion One system The saline prewash would be just lost time. It accomplishes nothing that will not be accomplished by the sucrose, which is equally effective at washing out the blood (more effective given the high pressure). Start with the sucrose. Time is of the essence, tissue deterioration begins immediately upon anoxia. Switch to fixative when the animal's muscles cease to move. 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 www.myneurolab.com -----Original Message----- From: Nancy.Walker@sanofi-synthelabo.com [mailto:Nancy.Walker@sanofi-synthelabo.com] Sent: Friday, October 03, 2003 8:12 AM To: Charles W. Scouten, Ph.D.; Histonet@lists.utsouthwestern.edu Subject: shrinkage and Perfusion One system Hello, >From the website mentioned in C. Scouten's mail I recuperated this info: The shrinkage occurs when a prewash with physiological saline is followed by fixative. The first action of fixative on the cell membrane is to shut off the sodium pump proteins. Sodium rushes into the cell, followed by water to maintain tonicity, and cells swell and expand. Membrane proteins are fixed and crosslinked to neighboring cells in the swollen position. Later, the cells stabilize and contract, and pull other cells with them. The result is gross shrinkage, local distortion and torn membranes with lost cellular contents from some cells. Tissue Shrinkage can be completely prevented if the extracellular fluid with ions is replaced by a nonionic isotonic solution that cannot enter the cells. This can be accomplished by prewash with 5% (isotonic) sucrose in distilled water. Start the prewash at low pressure, and pump up to 300 mmHg to break the blood brain barrier and wash the extracellular fluid (and red blood cells). Switch shortly thereafter to fixative. The perfusion takes the same time as the traditional procedure, and accomplishes the favorable results bulleted above. So if I'd like to try out the perfusion techniques that are made simple by the Perfusion one system, what would you suggest: first do a physiological saline solution, followed by a prewash with 5% sucrose and then PFA, or go directly to 5% sucrose then PFA? thanks for your advice, Nancy _______________________________________________ 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 -------------------------------------------------------------------- mail2web - Check your email from the web at http://mail2web.com/ . -------------------------------------------------------------------- mail2web - Check your email from the web at http://mail2web.com/ . From STEGTM <@t> samcstl.org Wed Oct 8 12:49:59 2003 From: STEGTM <@t> samcstl.org (Therersa Stegall) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Purple Haze..... Message-ID: Kathleen: Thanks for naming the amorphous blight that has been plaguing us lately. This "Jimi Hendrix" artifact has been running 'round our brains for the past few weeks. We have 1-started filtering the hematoxylin each morning, 2-alternated between Sta-on, gelatin, and naked waterbaths, 3-scrubbed the entire staining system, 4-decreased time in microwave, ovens, for drying slides; and yet, we still find this on some biopsies. Not always all the biopsies in a batch, this led us to ask GI about collection procedures, and possible air-drying, they insist it's not them. We've checked pH on water, accurately calibrated the bluing and acid/water washes (we stain regressively), and just about run out of ideas. I'm beginning to suspect the hematoxylin is leaving a precipitate, or maybe the microtomy is inconsistent and ruining cell integrity, or .... maybe it's terrorism! Any ideas out there? From georgecole <@t> ev1.net Wed Oct 8 12:57:37 2003 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Muscle and Nerve packets Message-ID: <000801c38dc5$aa553350$034dbad0@hppav> Folks, your response has been great. So far, I've sent out 50 Muscle and Nerve packets around the states and 14 to countries around the world. I can't help but wonder if these procedures will be 'an interesting' bunch of pages in the clutter of other such pages gathering dust in a heap of papers, or are some of these improvements getting on the line with histotechs doing them in their muscle and nerve biopsies? I'm not the pop quiz type----I'm the grandpop quiz kind----excuse me---this keyboard did it. I will just leave this little cluster of words hanging---- For so many years chasing after ways to find possibly life-saving information from nerve and muscle tissues, it became a stern rule that any---that is ANY---improvement in the effectiveness of the search would be adopted without regard for the comfort of old habits, the relative difficulty of the improved me, or the time it took to complete and adopt the improved procedures, although I would, of course, work to 'get going' with new, improved procedures. This became, please, pardon the purple prose, a MORAL IMPERATIVE. Processing a biopsy is not a histotech doing his or her thing----it's a histotech doing everything to the nth degree with the biopsy tissues for the good of the patient. "Some people do it that way, some people do it this way---ho hum" No. We ALL go ALL the way ALL THE TIME searching though those bits of people. Gad---an aria popped out----but that's it, I think----I've never seen a histotech become the darling of the paparazzi ---We wrestle in a lab with those tricky tissues to make them yield their secrets-BUT when the neuropathologist said to me, "We never would have found this condition if you hadn't done those procedures", that line from Gilbert and Sullivan played in my head: "I wouldn't trade places with Admiral Nelson hisself, no matter who he was a-huggin' of at the moment.". . No I realize I have sent packets out. That's it. What happens now is up to all you histotechs out there. However, I wouldn't mind at all if an occasional word, now and then, would show up on the histonet or on my e-mail about any of those procedures from the packet being adopted in your labs. I have more packets----they are waiting breathlessly to go out to into the world. georgecole@ev1.net -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/f7ea6b54/attachment.htm From mcauliff <@t> umdnj.edu Wed Oct 8 13:11:33 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Perfusion debate In-Reply-To: References: Message-ID: <3F845355.9040100@umdnj.edu> Joining in the perfusion debate......... I have perfused mouse and rat CNS with a pump and had excellent results. I have never measured the pressure the pump puts out but I do use a syringe needle (into the left ventricle) with a large diameter, 16 gauge for a rat, 20 gauge for a mouse. I think something close to the inside diameter of the aorta is required for good results. In my experience, high rate of flow is more important that pressure. I don't worry about washing out every last RBC, I don't think it can be done. I use a minimum amount of buffer (a few milleliters) as a prewash and I don't use vasodilators or anti-coagulents. No shrinkage problems for LM or EM. I have never tried sucrose as a prewash. I have also perfused rat kidney with just two jars high on a shelf above the sink. Beautiful EM's from that work, even from the renal medulla which has relatively low blood flow. Again, I did not measure the pressure but it was only about 90-100 mm Hg (converting the height in inches to mm then converting from water to mercury using specific gravity). To summarize, use a high rate of flow and be sure the buffer (not the fix+buffer solution) you use is somwhat hypertonic. This last point is well documented in the literature. Geoff mucram11@earthlink.net wrote: > Strange we did not have shrinkage problems with our set up. Pam Marcum > > Original Message: > ----------------- > From: Charles W. Scouten, Ph.D. cwscouten@myneurolab.com > Date: Wed, 8 Oct 2003 10:39:33 -0500 > To: mucram11@earthlink.net, frouwke@sci.kun.nl, > nancy.walker@sanofi-synthelabo.com, histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] RE: shrinkage and Perfusion One system > > > Yes, gravity will work, with 20% shrinkage of the brain, some shape > distortion, and with a reddish tinge due to remaining red blood cells, > which catalyze any HRP reaction, and autofluoresce. OK for some > purposes, > terrible for others, but not the optimum for any. > I have asked www.Histonet.org to post a picture called Perfused Brains > Annotated which shows whole rat brains, one fresh dissected out, one > perfused by pressure sucrose and 4% paraformaldehyde/glut, and the > last by > traditional gravity flow with saline prewash, and the same fixative. > > The last is reddish and shrunken compared to the middle brain. > Three ft of water translates directly, using the units converter at: > http://www.sciencemadesimple.com/conversions.html > to 67 mm Hg, below average diastolic pressure in mice or rats or > people. And some pressure is lost in the lines and needle. Even in > living animals > with cycles of systolic pressure, some capillaries occasionally get > plugged > for a while and stop flowing. That is part of the advantage of exercise, > to keep the lines open. It is extremely unlikely that completely open > exposure to 67 mm Hg would in any way "blow" the vascular system of any > mammal, or even clear out all the blood. The blocked passages will stop > fixative from reaching the surrounding tissue in that area, and lead to > local deterioration in unpredictable and irreproduceible areas. > > The only advantage of sucrose over saline is to clear the extracellular > fluid of sodium before fixative hits. High pressure, 300 mm Hg, > applied to > either saline or sucrose as the prewash, (there is never a reason for > both), will thoroughly clear all red blood cells, but should not be > pumped > up in advance the 'thump' the system, but increase over a few seconds to > clear most blood before the blood brain barrier opens. This prewash will > take half the time of traditional prewash, and get fixative in sooner and > more thoroughly. > > 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 www.myneurolab.com > > -----Original Message----- > From: mucram11@earthlink.net [mailto:mucram11@earthlink.net] Sent: > Wednesday, October 08, 2003 8:06 AM > To: frouwke@sci.kun.nl; Charles W. Scouten, Ph.D.; > nancy.walker@sanofi-synthelabo.com; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] RE: shrinkage and Perfusion One system > > Hi, > We never had a pump. We used gravity for over 2,000 rats so I think I can > say it will work. The height of the container or IV bottle used > seemed to > be the key. We controlled flow with the standard IV roller closure on > the > tubing. It was approximately 3ft above the area the animal was being > perfused in, always under a hood or very well ventilated area. We looked > for a steady drip in the tubing and out the cannula. We did not open the > valve all the way as it would blow out the vascular system. We did have a > couple of students try wide open and prove the point very well. > The use of a saline solution with and without anticoag agents had been > used. If you are going to use sucrose I would use the pre-wash to get > the > blood and red cells out. Just to be sure you have all the small vessels > cleared for the fixative or freezing technique. > Pam Marcum > > Original Message: > ----------------- > From: Frouwke Kuijpers frouwke@sci.kun.nl > Date: Wed, 8 Oct 2003 12:47:14 +0200 > To: cwscouten@myneurolab.com, Nancy.Walker@sanofi-synthelabo.com, > Histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] RE: shrinkage and Perfusion One system > > > Start the prewash at low pressure, and pump up to 300 mmHg > to break the blood brain barrier and wash the extracellular fluid (and > red > blood cells). > > Charles: next question: we don't do the perfusion by a pump, just by > gravity. You said the pressure for the 10% sucrose must go till > mm300Hg.... > Well we dont' have a pump: what will your advice be than? > Start with the saline solution to get rid of the blood and than the > sucrose? > > Frouwke Kuijpers > > ----- Original Message ----- From: "Charles W. Scouten, Ph.D." > > To: ; > > Sent: Monday, October 06, 2003 3:53 PM > Subject: [Histonet] RE: shrinkage and Perfusion One system > > > The saline prewash would be just lost time. It accomplishes nothing that > will not be accomplished by the sucrose, which is equally effective at > washing out the blood (more effective given the high pressure). Start > with > the sucrose. Time is of the essence, tissue deterioration begins > immediately upon anoxia. Switch to fixative when the animal's muscles > cease > to move. > > 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 > www.myneurolab.com > > > -----Original Message----- > From: Nancy.Walker@sanofi-synthelabo.com > [mailto:Nancy.Walker@sanofi-synthelabo.com] > Sent: Friday, October 03, 2003 8:12 AM > To: Charles W. Scouten, Ph.D.; Histonet@lists.utsouthwestern.edu > Subject: shrinkage and Perfusion One system > > > Hello, > >> From the website mentioned in C. Scouten's mail I recuperated this info: > > > The shrinkage occurs when a prewash with physiological saline is followed > by fixative. The first action of fixative on the cell membrane is to shut > off the sodium pump proteins. Sodium rushes into the cell, followed by > water to maintain tonicity, and cells swell and expand. Membrane proteins > are fixed and crosslinked to neighboring cells in the swollen position. > Later, the cells stabilize and contract, and pull other cells with them. > The result is gross shrinkage, local distortion and torn membranes with > lost cellular contents from some cells. > > Tissue Shrinkage can be completely prevented if the extracellular fluid > with ions is replaced by a nonionic isotonic solution that cannot > enter the > cells. This can be accomplished by prewash with 5% (isotonic) sucrose in > distilled water. Start the prewash at low pressure, and pump up to 300 > mmHg > to break the blood brain barrier and wash the extracellular fluid (and > red > blood cells). Switch shortly thereafter to fixative. The perfusion takes > the same time as the traditional procedure, and accomplishes the > favorable > results bulleted above. > > So if I'd like to try out the perfusion techniques that are made > simple by > the Perfusion one system, what would you suggest: first do a > physiological saline solution, followed by a prewash with 5% sucrose and > then PFA, or go directly to 5% sucrose then PFA? > > thanks for your advice, > > Nancy > > -- -- ********************************************** 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 ********************************************** -- -- ********************************************** 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 Salvacion.DeLovino <@t> cshs.org Wed Oct 8 13:26:30 2003 From: Salvacion.DeLovino <@t> cshs.org (DeLovino, Salvacion S.) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] freezing in isopentane Message-ID: What would be the price tag of that, if I may ask? Thank you! Salvacion Delovino -----Original Message----- From: Steven Slap [mailto:siksik@vgernet.net] Sent: Wednesday, October 08, 2003 7:25 AM To: HistoNet; Gayle Callis; Phil Oshel Subject: [Histonet] freezing in isopentane Hi HistoNetters At risk of someone writing "nice ad" again, there is an instrument, the "Clini-RF", from Bright Instruments, that would be very useful in resolving some of these issues. Instead of using liquid nitrogen to cool the isopentane, the Clini-RF is a stand-alone freezer with a -80 degree tank which can be used for direct immersion in isopentane. Although this instrument is available from Hacker Instruments in the United States, I get nothing except satisfaction from seeing it sold. best regards, Steven Slap _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ryaskovich <@t> dir.nidcr.nih.gov Wed Oct 8 13:48:11 2003 From: ryaskovich <@t> dir.nidcr.nih.gov (Yaskovich, Ruth A (NIH/NIDCR)) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Purple Haze..... Message-ID: <8F3AB322628548428A992EFB0E80F5D3A2EDDC@nihexchange8.nih.gov> I used to get this problem once in awhile and haven't had it since switching to charged slides. May-be in the amounts of gelatin? What kind of hematoxylin are you using? Ruth Yaskovich N.I.D.C.R. N.I.H. -----Original Message----- From: Therersa Stegall [mailto:STEGTM@samcstl.org] Sent: Wednesday, October 08, 2003 1:50 PM To: KMMerril@LancasterGeneral.org; histonet@pathology.swmed.edu Subject: [Histonet] Purple Haze..... Kathleen: Thanks for naming the amorphous blight that has been plaguing us lately. This "Jimi Hendrix" artifact has been running 'round our brains for the past few weeks. We have 1-started filtering the hematoxylin each morning, 2-alternated between Sta-on, gelatin, and naked waterbaths, 3-scrubbed the entire staining system, 4-decreased time in microwave, ovens, for drying slides; and yet, we still find this on some biopsies. Not always all the biopsies in a batch, this led us to ask GI about collection procedures, and possible air-drying, they insist it's not them. We've checked pH on water, accurately calibrated the bluing and acid/water washes (we stain regressively), and just about run out of ideas. I'm beginning to suspect the hematoxylin is leaving a precipitate, or maybe the microtomy is inconsistent and ruining cell integrity, or .... maybe it's terrorism! Any ideas out there? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Wed Oct 8 13:57:00 2003 From: cwscouten <@t> myneurolab.com (Charles W. Scouten, Ph.D.) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] freezing in isopentane Message-ID: The price tag for the Clini-RF from us is $5,239.00 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 www.myneurolab.com -----Original Message----- From: DeLovino, Salvacion S. [mailto:Salvacion.DeLovino@cshs.org] Sent: Wednesday, October 08, 2003 1:27 PM To: 'Steven Slap'; HistoNet; Gayle Callis; Phil Oshel Subject: RE: [Histonet] freezing in isopentane What would be the price tag of that, if I may ask? Thank you! Salvacion Delovino -----Original Message----- From: Steven Slap [mailto:siksik@vgernet.net] Sent: Wednesday, October 08, 2003 7:25 AM To: HistoNet; Gayle Callis; Phil Oshel Subject: [Histonet] freezing in isopentane Hi HistoNetters At risk of someone writing "nice ad" again, there is an instrument, the "Clini-RF", from Bright Instruments, that would be very useful in resolving some of these issues. Instead of using liquid nitrogen to cool the isopentane, the Clini-RF is a stand-alone freezer with a -80 degree tank which can be used for direct immersion in isopentane. Although this instrument is available from Hacker Instruments in the United States, I get nothing except satisfaction from seeing it sold. best regards, Steven Slap _______________________________________________ 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 cwscouten <@t> myneurolab.com Wed Oct 8 14:06:15 2003 From: cwscouten <@t> myneurolab.com (Charles W. Scouten, Ph.D.) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] RE: shrinkage and Perfusion One system Message-ID: I didn't know the brains I was working with had shrunk until I used the pressure sucrose perfusion. Suddenly I couldn't get two coronal sections through the hamster brain on the same slide if arranged side by side, across the width. I had been doing that, but had to rotate my placement after using the new perfusion. It is a very old and well known observation, documented in the Atlas of Konig and Klippel, and the reason Paxinos chooses to work only with fresh unfixed tissue, in spite of the hassels. Total loss of the extracellular space is also documented, most EM people working with brain are familiar with the problem, some use the pressure sucrose method devised by Cragg, from whom I got the idea. 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 www.myneurolab.com -----Original Message----- From: mucram11@earthlink.net [mailto:mucram11@earthlink.net] Sent: Wednesday, October 08, 2003 12:20 PM To: Charles W. Scouten, Ph.D.; mucram11@earthlink.net; frouwke@sci.kun.nl; nancy.walker@sanofi-synthelabo.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: shrinkage and Perfusion One system Strange we did not have shrinkage problems with our set up. Pam Marcum Original Message: ----------------- From: Charles W. Scouten, Ph.D. cwscouten@myneurolab.com Date: Wed, 8 Oct 2003 10:39:33 -0500 To: mucram11@earthlink.net, frouwke@sci.kun.nl, nancy.walker@sanofi-synthelabo.com, histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: shrinkage and Perfusion One system Yes, gravity will work, with 20% shrinkage of the brain, some shape distortion, and with a reddish tinge due to remaining red blood cells, which catalyze any HRP reaction, and autofluoresce. OK for some purposes, terrible for others, but not the optimum for any. I have asked www.Histonet.org to post a picture called Perfused Brains Annotated which shows whole rat brains, one fresh dissected out, one perfused by pressure sucrose and 4% paraformaldehyde/glut, and the last by traditional gravity flow with saline prewash, and the same fixative. The last is reddish and shrunken compared to the middle brain. Three ft of water translates directly, using the units converter at: http://www.sciencemadesimple.com/conversions.html to 67 mm Hg, below average diastolic pressure in mice or rats or people. And some pressure is lost in the lines and needle. Even in living animals with cycles of systolic pressure, some capillaries occasionally get plugged for a while and stop flowing. That is part of the advantage of exercise, to keep the lines open. It is extremely unlikely that completely open exposure to 67 mm Hg would in any way "blow" the vascular system of any mammal, or even clear out all the blood. The blocked passages will stop fixative from reaching the surrounding tissue in that area, and lead to local deterioration in unpredictable and irreproduceible areas. The only advantage of sucrose over saline is to clear the extracellular fluid of sodium before fixative hits. High pressure, 300 mm Hg, applied to either saline or sucrose as the prewash, (there is never a reason for both), will thoroughly clear all red blood cells, but should not be pumped up in advance the 'thump' the system, but increase over a few seconds to clear most blood before the blood brain barrier opens. This prewash will take half the time of traditional prewash, and get fixative in sooner and more thoroughly. 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 www.myneurolab.com -----Original Message----- From: mucram11@earthlink.net [mailto:mucram11@earthlink.net] Sent: Wednesday, October 08, 2003 8:06 AM To: frouwke@sci.kun.nl; Charles W. Scouten, Ph.D.; nancy.walker@sanofi-synthelabo.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: shrinkage and Perfusion One system Hi, We never had a pump. We used gravity for over 2,000 rats so I think I can say it will work. The height of the container or IV bottle used seemed to be the key. We controlled flow with the standard IV roller closure on the tubing. It was approximately 3ft above the area the animal was being perfused in, always under a hood or very well ventilated area. We looked for a steady drip in the tubing and out the cannula. We did not open the valve all the way as it would blow out the vascular system. We did have a couple of students try wide open and prove the point very well. The use of a saline solution with and without anticoag agents had been used. If you are going to use sucrose I would use the pre-wash to get the blood and red cells out. Just to be sure you have all the small vessels cleared for the fixative or freezing technique. Pam Marcum Original Message: ----------------- From: Frouwke Kuijpers frouwke@sci.kun.nl Date: Wed, 8 Oct 2003 12:47:14 +0200 To: cwscouten@myneurolab.com, Nancy.Walker@sanofi-synthelabo.com, Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: shrinkage and Perfusion One system Start the prewash at low pressure, and pump up to 300 mmHg to break the blood brain barrier and wash the extracellular fluid (and red blood cells). Charles: next question: we don't do the perfusion by a pump, just by gravity. You said the pressure for the 10% sucrose must go till mm300Hg.... Well we dont' have a pump: what will your advice be than? Start with the saline solution to get rid of the blood and than the sucrose? Frouwke Kuijpers ----- Original Message ----- From: "Charles W. Scouten, Ph.D." To: ; Sent: Monday, October 06, 2003 3:53 PM Subject: [Histonet] RE: shrinkage and Perfusion One system The saline prewash would be just lost time. It accomplishes nothing that will not be accomplished by the sucrose, which is equally effective at washing out the blood (more effective given the high pressure). Start with the sucrose. Time is of the essence, tissue deterioration begins immediately upon anoxia. Switch to fixative when the animal's muscles cease to move. 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 www.myneurolab.com -----Original Message----- From: Nancy.Walker@sanofi-synthelabo.com [mailto:Nancy.Walker@sanofi-synthelabo.com] Sent: Friday, October 03, 2003 8:12 AM To: Charles W. Scouten, Ph.D.; Histonet@lists.utsouthwestern.edu Subject: shrinkage and Perfusion One system Hello, >From the website mentioned in C. Scouten's mail I recuperated this info: The shrinkage occurs when a prewash with physiological saline is followed by fixative. The first action of fixative on the cell membrane is to shut off the sodium pump proteins. Sodium rushes into the cell, followed by water to maintain tonicity, and cells swell and expand. Membrane proteins are fixed and crosslinked to neighboring cells in the swollen position. Later, the cells stabilize and contract, and pull other cells with them. The result is gross shrinkage, local distortion and torn membranes with lost cellular contents from some cells. Tissue Shrinkage can be completely prevented if the extracellular fluid with ions is replaced by a nonionic isotonic solution that cannot enter the cells. This can be accomplished by prewash with 5% (isotonic) sucrose in distilled water. Start the prewash at low pressure, and pump up to 300 mmHg to break the blood brain barrier and wash the extracellular fluid (and red blood cells). Switch shortly thereafter to fixative. The perfusion takes the same time as the traditional procedure, and accomplishes the favorable results bulleted above. So if I'd like to try out the perfusion techniques that are made simple by the Perfusion one system, what would you suggest: first do a physiological saline solution, followed by a prewash with 5% sucrose and then PFA, or go directly to 5% sucrose then PFA? thanks for your advice, Nancy _______________________________________________ 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 -------------------------------------------------------------------- mail2web - Check your email from the web at http://mail2web.com/ . -------------------------------------------------------------------- mail2web - Check your email from the web at http://mail2web.com/ . From JBurrill <@t> criver.com Wed Oct 8 14:27:49 2003 From: JBurrill <@t> criver.com (JBurrill@criver.com) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] TNFalpha staining on mouse tissue Message-ID: Has anyone had any luck staining for TNFalpha in formalin fixed paraffin embedded mouse tissue sections. Thanks in advance. Jason Jason D. Burrill, B.A., HT(ASCP) Senior Supervisor, Histology Charles River Laboratories 251 Ballardvale Street Wilmington, MA 01887 ph: 978-658-6000 ext 1652 fax: 978-988-8793 E-mail: jburrill@criver.com -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/5b4d03dc/attachment.htm From NDimaano <@t> howost.com Wed Oct 8 14:34:33 2003 From: NDimaano <@t> howost.com (Dimaano, Nena) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Journal of Histotechnology Message-ID: <20BE9059B6FC2D4B9E8FF2A9E2C4EEBD13F20C@HOS2KEXCHCL.howost.strykercorp.com> Skipped content of type multipart/alternative From jschuma1 <@t> FAIRVIEW.ORG Wed Oct 8 15:35:13 2003 From: jschuma1 <@t> FAIRVIEW.ORG (JENNIFER SCHUMACHER) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Sentinel nodes Message-ID: Good afternoon everyone, We routinely cut our negative sentinel node biopsies shallow, cutting 5 levels (mounting levels 2 and 4 for IHC), approximately 30 microns between levels. Now, a pathologist read that "most institutions" are cutting them deeper, approximately 100 microns between levels. This is totally an INFORMAL poll, but I would appreciate any feedback from the community. I will check the archives as well, but thought I would get the question out there before everyone heads off to Kentucky:-) Thanks. Jennifer -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/bab5b703/attachment.htm From vermast <@t> rogers.com Wed Oct 8 15:56:37 2003 From: vermast <@t> rogers.com (vermast) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] negative IHC controls Message-ID: <000f01c38dde$aa247d00$57c08a3f@yoursz6x6sefxo> I would like to get a feel for how many out there are running negative control slides for IHC. In our lab we do just a handful of antibodies and initially I had been running a negative control slide with each patient slide. After much discussion with our pathologists, we decided to omit these negatives (which were conistently negative) and continue to just run a positive control with each primary antibody for the run. We use the Dako autostainer and prediluted primaries. The decision to stop running negatives also coincided with Dako's decision to sell the negative control sera separately from the primaries (they used to come packaged together). Perhaps I assumed that discontinuing to pair these reagents together meant that few labs were using the negatives. Anyhow, after having reviewed the last QMPLS (Canada) survey committee comments, I believe the committe would like a negative control run with each patient tissue slide in order to evaluate background (they have used NCCLS guide pages as reference). Incidentally we weren't a part of the survey due to a technicality. Any help or advice would be appreciated. L. Vermast Stratford, Ont. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/9cd6f406/attachment.htm From Dren6289 <@t> aol.com Wed Oct 8 16:05:29 2003 From: Dren6289 <@t> aol.com (Dren6289@aol.com) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Question On Microwave processing Message-ID: <16.3637398c.2cb5d619@aol.com> If you use microwave for tissue processing, can you please e-mail me back with the reason you use it and the benefits. I need some ammunition for the boss. Thanks -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/6c9b1e8a/attachment.htm From nick.kirk3 <@t> btopenworld.com Wed Oct 8 16:30:10 2003 From: nick.kirk3 <@t> btopenworld.com (Nick Kirk) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Cost In-Reply-To: Message-ID: Jenny We have a fairly robust method of costing where the total cost is broken down into two component parts, the staff cost and the variable or consumables cost. The staff cost is worked out on a pro-rata basis for a mid point on a basic grade Biomedical Scientist's salary, working out an average time taken to do the particular procedure and then costed according to that hourly rate. The variable costs are simply broken down to the actual costs of each individual item in that process, i.e. the cost of an individual slide, the cost of the particular number of micro litres of an immuno reagent required etc. As it's broken down this way it makes it much easier to adjust the costs charged to the "customer", as the costs of the individual items within that process change with time. It's a bit of work initially but it is very easy to manage once set up. I use an Excel spreadsheet which does all the calculations for me and I can charge the "customer" accordingly. Obviously charges will vary enormously depending on where you are in the world, but the principal of the system should be the same no matter where you are. Tot siens Geniet jou dag Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Jenny Molde Sent: 08 October 2003 08:35 To: histonet@pathology.swmed.edu Subject: [Histonet] Cost Hi everyone out there I would like to know more or less what one would charge for a routine H+E and special stain on a sample recieved from explant and embedded in MMA resin, cut and stained. Also the immunocytochemical charge please. All suggestions will be greatly appreciated. Many thanks. Jenny Molde Cardiovascular Research Unit University of Cape Town South Africa. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nick.kirk3 <@t> btopenworld.com Wed Oct 8 16:30:10 2003 From: nick.kirk3 <@t> btopenworld.com (Nick Kirk) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Purple Haze..... In-Reply-To: Message-ID: If this artefact is what I think you are talking about and that is a general non-descript blue haze in the nuclei where chromatin detail is non-existent, we in the UK have also experienced much the same problem. Some years ago this problem lead to a national study being done through the auspices of our National External Quality Control organisation (NEQAS). If I remember correctly (and I'm sure someone over here will correct me if I'm wrong) no firm conclusions were made as to what causes this artefact (in the UK we often call this "blue blob syndrome") Basically the nuclear meltdown artefact, to give it another name, was principally found in endoscopic biopsies, usually of the GI tract, but was occasionally seen in other tissue as well. I personally think it's a multi-factorial phenomenon and has something to do with how the biopsy is taken and the length of time it takes for the biopsy to reach fixative from the endoscope. Coupled together with other as yet unknown factors, it produces the said artefact. That is why it is an intermittent problem. I do seem to recall now that I think a bit, that there was some link postulated with vacuum processing as well, but I don't think it was proven and as I said earlier, this is an intermittent phenomenon so I don't think that's the answer either. We'll probably never know. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England. -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Therersa Stegall Sent: 08 October 2003 18:50 To: KMMerril@LancasterGeneral.org; histonet@pathology.swmed.edu Subject: [Histonet] Purple Haze..... Kathleen: Thanks for naming the amorphous blight that has been plaguing us lately. This "Jimi Hendrix" artifact has been running 'round our brains for the past few weeks. We have 1-started filtering the hematoxylin each morning, 2-alternated between Sta-on, gelatin, and naked waterbaths, 3-scrubbed the entire staining system, 4-decreased time in microwave, ovens, for drying slides; and yet, we still find this on some biopsies. Not always all the biopsies in a batch, this led us to ask GI about collection procedures, and possible air-drying, they insist it's not them. We've checked pH on water, accurately calibrated the bluing and acid/water washes (we stain regressively), and just about run out of ideas. I'm beginning to suspect the hematoxylin is leaving a precipitate, or maybe the microtomy is inconsistent and ruining cell integrity, or .... maybe it's terrorism! Any ideas out there? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LuckG <@t> empirehealth.org Wed Oct 8 16:35:08 2003 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Sentinel nodes Message-ID: Jennifer, We cut levels x 3 on all negative sentinel lymph node blocks. Levels 1 & 3 become H&E's and two #2 levels are held for possible IHC's. We rough cut into the block approximately 1/3 of it's depth and pickup the 1st level. The 2nd levels are at mid block and the third level is at the deeper 2/3's of the block. I guess in answer to your question about intervals between levels, here it is determined relative to the depth of the tissue in the paraffin block. Greg Luck Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 509.473.7077 luckg@empirehealth.org -----Original Message----- From: JENNIFER SCHUMACHER [mailto:jschuma1@FAIRVIEW.ORG] Sent: Wednesday, October 08, 2003 1:35 PM To: histonet@lists.utsouthwestern.EDU Subject: [Histonet] Sentinel nodes Good afternoon everyone, We routinely cut our negative sentinel node biopsies shallow, cutting 5 levels (mounting levels 2 and 4 for IHC), approximately 30 microns between levels. Now, a pathologist read that "most institutions" are cutting them deeper, approximately 100 microns between levels. This is totally an INFORMAL poll, but I would appreciate any feedback from the community. I will check the archives as well, but thought I would get the question out there before everyone heads off to Kentucky:-) Thanks. Jennifer -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/3af7a038/attachment.htm From cwscouten <@t> myneurolab.com Wed Oct 8 17:09:11 2003 From: cwscouten <@t> myneurolab.com (Charles W. Scouten, Ph.D.) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Perfusion debate Message-ID: The large diameter needle into the ventricle sounds great. High flow rate sounds good to get clean of red blood cells, but how high is high, and in what species and body size? Of course, pressure and flow rate are closely related, the third determining variable is cardiovascular resistance, which is related to size, gender, species, health condition, etc. A mouse would have far more resistance to a given flow than would a pig. However, as might be guessed, all mammals have very close to the same average blood pressure. See the table below, (pointed out to me by histonetters, thank you again): The following is a table of average blood pressure for several species (systolic/diastolic) (Green, 1979). mice 113/81 rats 116/90 hamster 150/110 rabbit 110/80 dog 112/56 cat 120/75 baboon 148/100 rhesus 160/127 pig 170/108. Consider what would happen if you perfused a pig with the same flow rate that works well on a mouse, or vice versa. But you can use the same pressure to perfuse a pig or mouse, and get comparable results. Controlling pressure is safer and more predictable. A high flow rate means high pressure, but the operational definition of high flow rate must depend on the species and body size, among other variables. High but safe pressure is 200 mm Hg in all of the above. The blood brain barrier is reliable broken in hamsters and rats at 300 mm Hg. What do you bet for the rest? At 300 mm Hg, I come very close to washing out every last RBC in Hamster or Rat brain. HRP shows maybe one in each whole brain coronal 60 micron thick section. There is no noticeable (by me) damage to capillaries, but maybe a better anatomist could see it. You do have shrinkage in the CNS, it is well documented. You don't see it as a problem because it has always been there, and you haven't worked with fixed sections without it. See Konig and Klippel for "inevitable shrinkage", and ask Paxinos why his atlases are done only on fresh, not fixed tissue. But you can have both fixed and no shrinkage. You EM sections showed little or no extracellular space. Actually, the 5% sucrose is slightly hypertonic, isotonic is something like 4.94 by weight. I had never heard that point, though, can you tell me more about the documentation on that? Kidney is different, there is no blood brain barrier, and near isotonic sucrose would replace the extracellular fluid without the high pressure. Isn't the active sodium pump limited to only exciteable cells? If so, this formalin shrinkage thing would only be relevant for neural or muscle cells. Does anybody know if kidney or liver, for example, shrink with perfusion? 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 www.myneurolab.com -----Original Message----- From: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] Sent: Wednesday, October 08, 2003 1:12 PM To: histonet@pathology.swmed.edu Subject: Re: [Histonet] Perfusion debate Joining in the perfusion debate......... I have perfused mouse and rat CNS with a pump and had excellent results. I have never measured the pressure the pump puts out but I do use a syringe needle (into the left ventricle) with a large diameter, 16 gauge for a rat, 20 gauge for a mouse. I think something close to the inside diameter of the aorta is required for good results. In my experience, high rate of flow is more important that pressure. I don't worry about washing out every last RBC, I don't think it can be done. I use a minimum amount of buffer (a few milleliters) as a prewash and I don't use vasodilators or anti-coagulents. No shrinkage problems for LM or EM. I have never tried sucrose as a prewash. I have also perfused rat kidney with just two jars high on a shelf above the sink. Beautiful EM's from that work, even from the renal medulla which has relatively low blood flow. Again, I did not measure the pressure but it was only about 90-100 mm Hg (converting the height in inches to mm then converting from water to mercury using specific gravity). To summarize, use a high rate of flow and be sure the buffer (not the fix+buffer solution) you use is somwhat hypertonic. This last point is well documented in the literature. Geoff mucram11@earthlink.net wrote: > Strange we did not have shrinkage problems with our set up. Pam Marcum > > Original Message: > ----------------- > From: Charles W. Scouten, Ph.D. cwscouten@myneurolab.com > Date: Wed, 8 Oct 2003 10:39:33 -0500 > To: mucram11@earthlink.net, frouwke@sci.kun.nl, > nancy.walker@sanofi-synthelabo.com, histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] RE: shrinkage and Perfusion One system > > > Yes, gravity will work, with 20% shrinkage of the brain, some shape > distortion, and with a reddish tinge due to remaining red blood cells, > which catalyze any HRP reaction, and autofluoresce. OK for some > purposes, > terrible for others, but not the optimum for any. > I have asked www.Histonet.org to post a picture called Perfused Brains > Annotated which shows whole rat brains, one fresh dissected out, one > perfused by pressure sucrose and 4% paraformaldehyde/glut, and the > last by > traditional gravity flow with saline prewash, and the same fixative. > > The last is reddish and shrunken compared to the middle brain. > Three ft of water translates directly, using the units converter at: > http://www.sciencemadesimple.com/conversions.html > to 67 mm Hg, below average diastolic pressure in mice or rats or > people. And some pressure is lost in the lines and needle. Even in > living animals > with cycles of systolic pressure, some capillaries occasionally get > plugged > for a while and stop flowing. That is part of the advantage of exercise, > to keep the lines open. It is extremely unlikely that completely open > exposure to 67 mm Hg would in any way "blow" the vascular system of any > mammal, or even clear out all the blood. The blocked passages will stop > fixative from reaching the surrounding tissue in that area, and lead to > local deterioration in unpredictable and irreproduceible areas. > > The only advantage of sucrose over saline is to clear the extracellular > fluid of sodium before fixative hits. High pressure, 300 mm Hg, > applied to > either saline or sucrose as the prewash, (there is never a reason for > both), will thoroughly clear all red blood cells, but should not be > pumped > up in advance the 'thump' the system, but increase over a few seconds to > clear most blood before the blood brain barrier opens. This prewash will > take half the time of traditional prewash, and get fixative in sooner and > more thoroughly. > > 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 www.myneurolab.com > > -----Original Message----- > From: mucram11@earthlink.net [mailto:mucram11@earthlink.net] Sent: > Wednesday, October 08, 2003 8:06 AM > To: frouwke@sci.kun.nl; Charles W. Scouten, Ph.D.; > nancy.walker@sanofi-synthelabo.com; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] RE: shrinkage and Perfusion One system > > Hi, > We never had a pump. We used gravity for over 2,000 rats so I think I can > say it will work. The height of the container or IV bottle used > seemed to > be the key. We controlled flow with the standard IV roller closure on > the > tubing. It was approximately 3ft above the area the animal was being > perfused in, always under a hood or very well ventilated area. We looked > for a steady drip in the tubing and out the cannula. We did not open the > valve all the way as it would blow out the vascular system. We did have a > couple of students try wide open and prove the point very well. > The use of a saline solution with and without anticoag agents had been > used. If you are going to use sucrose I would use the pre-wash to get > the > blood and red cells out. Just to be sure you have all the small vessels > cleared for the fixative or freezing technique. > Pam Marcum > > Original Message: > ----------------- > From: Frouwke Kuijpers frouwke@sci.kun.nl > Date: Wed, 8 Oct 2003 12:47:14 +0200 > To: cwscouten@myneurolab.com, Nancy.Walker@sanofi-synthelabo.com, > Histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] RE: shrinkage and Perfusion One system > > > Start the prewash at low pressure, and pump up to 300 mmHg > to break the blood brain barrier and wash the extracellular fluid (and > red > blood cells). > > Charles: next question: we don't do the perfusion by a pump, just by > gravity. You said the pressure for the 10% sucrose must go till > mm300Hg.... > Well we dont' have a pump: what will your advice be than? > Start with the saline solution to get rid of the blood and than the > sucrose? > > Frouwke Kuijpers > > ----- Original Message ----- From: "Charles W. Scouten, Ph.D." > > To: ; > > Sent: Monday, October 06, 2003 3:53 PM > Subject: [Histonet] RE: shrinkage and Perfusion One system > > > The saline prewash would be just lost time. It accomplishes nothing that > will not be accomplished by the sucrose, which is equally effective at > washing out the blood (more effective given the high pressure). Start > with > the sucrose. Time is of the essence, tissue deterioration begins > immediately upon anoxia. Switch to fixative when the animal's muscles > cease > to move. > > 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 > www.myneurolab.com > > > -----Original Message----- > From: Nancy.Walker@sanofi-synthelabo.com > [mailto:Nancy.Walker@sanofi-synthelabo.com] > Sent: Friday, October 03, 2003 8:12 AM > To: Charles W. Scouten, Ph.D.; Histonet@lists.utsouthwestern.edu > Subject: shrinkage and Perfusion One system > > > Hello, > >> From the website mentioned in C. Scouten's mail I recuperated this info: > > > The shrinkage occurs when a prewash with physiological saline is followed > by fixative. The first action of fixative on the cell membrane is to shut > off the sodium pump proteins. Sodium rushes into the cell, followed by > water to maintain tonicity, and cells swell and expand. Membrane proteins > are fixed and crosslinked to neighboring cells in the swollen position. > Later, the cells stabilize and contract, and pull other cells with them. > The result is gross shrinkage, local distortion and torn membranes with > lost cellular contents from some cells. > > Tissue Shrinkage can be completely prevented if the extracellular fluid > with ions is replaced by a nonionic isotonic solution that cannot > enter the > cells. This can be accomplished by prewash with 5% (isotonic) sucrose in > distilled water. Start the prewash at low pressure, and pump up to 300 > mmHg > to break the blood brain barrier and wash the extracellular fluid (and > red > blood cells). Switch shortly thereafter to fixative. The perfusion takes > the same time as the traditional procedure, and accomplishes the > favorable > results bulleted above. > > So if I'd like to try out the perfusion techniques that are made > simple by > the Perfusion one system, what would you suggest: first do a > physiological saline solution, followed by a prewash with 5% sucrose and > then PFA, or go directly to 5% sucrose then PFA? > > thanks for your advice, > > Nancy > > -- -- ********************************************** 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 ********************************************** -- -- ********************************************** 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 nick.kirk3 <@t> btopenworld.com Wed Oct 8 17:22:59 2003 From: nick.kirk3 <@t> btopenworld.com (Nick Kirk) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] negative IHC controls In-Reply-To: <000f01c38dde$aa247d00$57c08a3f@yoursz6x6sefxo> Message-ID: In my opinion you should always run a negative control with every patient run. For starters, how do you know a. Your blocking solutions have worked? b. You haven't got endogenous biotin activity (especially in liver biopsies) giving a false positive? c. You haven't got a contaminant? For clinical assurance and governance reasons alone I would have thought that negative controls are essential. I think the reasons for DAKO selling the two reagents separate is for commercial reasons only (they can charge more) rather than any fact that people aren't using them. In the UK it is considered "best practice" to use negative controls and not to do so would be frowned upon by the National External Quality Assurance Scheme (NEQAS) who monitor the quality of immuno carried out by hospitals in the UK, which nearly every lab in the UK and many more overseas are signed up to. No negative controls is poor quality control in my book. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of vermast Sent: 08 October 2003 21:57 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] negative IHC controls I would like to get a feel for how many out there are running negative control slides for IHC. In our lab we do just a handful of antibodies and initially I had been running a negative control slide with each patient slide. After much discussion with our pathologists, we decided to omit these negatives (which were conistently negative) and continue to just run a positive control with each primary antibody for the run. We use the Dako autostainer and prediluted primaries. The decision to stop running negatives also coincided with Dako's decision to sell the negative control sera separately from the primaries (they used to come packaged together). Perhaps I assumed that discontinuing to pair these reagents together meant that few labs were using the negatives. Anyhow, after having reviewed the last QMPLS (Canada) survey committee comments, I believe the committe would like a negative control run with each patient tissue slide in order to evaluate background (they have used NCCLS guide pages as reference). Incidentally we weren't a part of the survey due to a technicality. Any help or advice would be appreciated. L. Vermast Stratford, Ont. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/84d45cbe/attachment.htm From cfarm <@t> sio.midco.net Wed Oct 8 17:23:06 2003 From: cfarm <@t> sio.midco.net (celia) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] warthin starry Message-ID: <000801c38dea$bf4500e0$cacb0945@youru10ixi0anw> Hello Everyone, I have put this question out once before and I didn't get much for a response so I thought I would try putting it out there one more time. I have an ongoing problem with my Warthin-Starry stain. The peripheral edges of some of the gastric biopsies are not staining. I am also experiencing this same non-staining with the Steiner stain on the Ventana stainer. I have tried reducing the temp on the processor down from 60c to 58c thinking they might be "cooking" on the processor (the H&E's look fine) as well as checking the ph of my solutions. This did not work. Could this be a handling problem prior to fixation in 10% NBF? I am at my wits end. Any suggestions will be greatly appreciated. Thanks in advance. Celia, HT SVH, Sioux Falls, SD -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/f2cd54e7/attachment.htm From nick.kirk3 <@t> btopenworld.com Wed Oct 8 17:38:25 2003 From: nick.kirk3 <@t> btopenworld.com (Nick Kirk) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Cost In-Reply-To: <3F8483D6.2050407@pathology.washington.edu> Message-ID: Yes, all such things as power, water etc come under a heading of capital charges which is incorporated into the staff costs part of the overall cost. The cost of doing the billing is free (we're so generous!) but it takes very little of my time with the spreadsheet I use so it doesn't amount to much anyway. (we're also grossly underpaid here in the UK) Nick -----Original Message----- From: Victor Tobias [mailto:victor@pathology.washington.edu] Sent: 08 October 2003 22:39 To: Nick Kirk Subject: Re: [Histonet] Cost Nick, Do you consider the cost of electricity, building lease, salary of the person actually submitting the billing, etc, into your cost? We call it overhead here in the states. Victor Nick Kirk wrote: >Jenny > >We have a fairly robust method of costing where the total cost is broken >down into two component parts, the staff cost and the variable or >consumables cost. >The staff cost is worked out on a pro-rata basis for a mid point on a basic >grade Biomedical Scientist's salary, working out an average time taken to do >the particular procedure and then costed according to that hourly rate. >The variable costs are simply broken down to the actual costs of each >individual item in that process, i.e. the cost of an individual slide, the >cost of the particular number of micro litres of an immuno reagent required >etc. >As it's broken down this way it makes it much easier to adjust the costs >charged to the "customer", as the costs of the individual items within that >process change with time. > >It's a bit of work initially but it is very easy to manage once set up. >I use an Excel spreadsheet which does all the calculations for me and I can >charge the "customer" accordingly. > >Obviously charges will vary enormously depending on where you are in the >world, but the principal of the system should be the same no matter where >you are. > >Tot siens >Geniet jou dag > >Nick Kirk >Histopathology >Hinchingbrooke Hospital >Huntingdon >England > > >-----Original Message----- >From: histonet-admin@lists.utsouthwestern.edu >[mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Jenny Molde >Sent: 08 October 2003 08:35 >To: histonet@pathology.swmed.edu >Subject: [Histonet] Cost > > >Hi everyone out there >I would like to know more or less what one would charge for a >routine H+E and special stain on a sample recieved from explant and >embedded in MMA resin, cut and stained. Also the immunocytochemical >charge please. All suggestions will be greatly appreciated. Many >thanks. > >Jenny Molde >Cardiovascular Research Unit >University of Cape Town >South Africa. > >_______________________________________________ >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 damianmole <@t> ntlworld.com Wed Oct 8 17:39:06 2003 From: damianmole <@t> ntlworld.com (damianmole) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] unsubscribe References: Message-ID: <005101c38dec$fe89d530$4d24fe3e@officedesk> From gliuygao <@t> hotmail.com Wed Oct 8 17:42:37 2003 From: gliuygao <@t> hotmail.com (yan gao) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Cut coronary artery Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/a28ee081/attachment.htm From Vfierke <@t> SNBLUSA.com Wed Oct 8 17:57:11 2003 From: Vfierke <@t> SNBLUSA.com (Vaughn Fierke) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Research histology opening Message-ID: There is an immediate opening for an HT or HTL in our Pathology Services at SNBL USA, Ltd pharmaceutical toxicology research center in Everett, Washington. Candidate would work primarily in immunohistochemistry, some EM processing and routine histology. Person should be comfortable designing study specific tests with four pathologists, able to work in a team atmosphere and communicate effectively with other lab staff. Technicians are trained in in rodent and NHP necropsy and trimming procedures. We offer competitve salary and benefits DOE. Our location is about 40 minutes north from downtown Seattle and next door to the Boeing 747-777 assembly area. Puget Sound shoreline is a few minutes to the west and within an hour to excellent skiing and hiking in the Cascade mountains to the east. For more information, check the website . Send resume to the human resources department or call 425-407-0121 ex2479 directly to the histology lab. Confidentiality Notice: This email, its contents and attachments are confidential and may contain privileged information. It is intended solely for the use of addressee(s) only. Any use, copying or disclosure of this communication or attachments to any other person is expressly prohibited without written permission of SNBL-USA,Ltd. If you receive this message in error, please notify the sender at SNBL USA, Ltd. immediately by return e-mail, telephone +1 425 407 0121, or fax +1 425 407 8601. We appreciate your cooperation From la.sebree <@t> hosp.wisc.edu Wed Oct 8 18:31:43 2003 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] negative IHC controls Message-ID: L.Vermast, We run a negative control with each patient slide/antibody. In some cases we can use a negative control for more than one antibody on the same patient IF the antibodies are from the same source, ie rabbit, mouse, ascites fluid, and if the staining protocols are identical. We also have statements from several of our pathologists that if their case has multiple blocks, one negative/antibody/CASE will suffice rather than one negative/antibody/BLOCK. We've never run into a problem doing it this way. -----Original Message----- From: vermast [mailto:vermast@rogers.com] Sent: Wed 10/8/2003 3:56 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] negative IHC controls I would like to get a feel for how many out there are running negative control slides for IHC. In our lab we do just a handful of antibodies and initially I had been running a negative control slide with each patient slide. After much discussion with our pathologists, we decided to omit these negatives (which were conistently negative) and continue to just run a positive control with each primary antibody for the run. We use the Dako autostainer and prediluted primaries. The decision to stop running negatives also coincided with Dako's decision to sell the negative control sera separately from the primaries (they used to come packaged together). Perhaps I assumed that discontinuing to pair these reagents together meant that few labs were using the negatives. Anyhow, after having reviewed the last QMPLS (Canada) survey committee comments, I believe the committe would like a negative control run with each patient tissue slide in order to evaluate background (they have used NCCLS guide pages as reference). Incidentally we weren't a part of the survey due to a technicality. Any help or advice would be appreciated. L. Vermast Stratford, Ont. From bhewlett <@t> cogeco.ca Wed Oct 8 19:04:51 2003 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] negative IHC controls References: <000f01c38dde$aa247d00$57c08a3f@yoursz6x6sefxo> Message-ID: <006701c38df9$07371790$6400a8c0@bryaniwx13voft> Negative reagent controls are supposed to be consistently negative. That's the point! The notion of not performing negative reagent controls on patient samples for IHC, because they have always been consistently negative in the past, is equivalent to the equally bizarre notion of removing the airbags, seat belts, child restraints and other safety equipment from your automobile, because you have not yet had an accident and have therefore never used them. Bryan Hewlett Consultant Technologist QMP-LS ----- Original Message ----- From: vermast To: histonet@lists.utsouthwestern.edu Sent: Wednesday, October 08, 2003 4:56 PM Subject: [Histonet] negative IHC controls I would like to get a feel for how many out there are running negative control slides for IHC. In our lab we do just a handful of antibodies and initially I had been running a negative control slide with each patient slide. After much discussion with our pathologists, we decided to omit these negatives (which were conistently negative) and continue to just run a positive control with each primary antibody for the run. We use the Dako autostainer and prediluted primaries. The decision to stop running negatives also coincided with Dako's decision to sell the negative control sera separately from the primaries (they used to come packaged together). Perhaps I assumed that discontinuing to pair these reagents together meant that few labs were using the negatives. Anyhow, after having reviewed the last QMPLS (Canada) survey committee comments, I believe the committe would like a negative control run with each patient tissue slide in order to evaluate background (they have used NCCLS guide pages as reference). Incidentally we weren't a part of the survey due to a technicality. Any help or advice would be appreciated. L. Vermast Stratford, Ont. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/1bbe4dbd/attachment.htm From GREYTRUNK <@t> aol.com Wed Oct 8 21:29:18 2003 From: GREYTRUNK <@t> aol.com (GREYTRUNK@aol.com) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Purple Haze..... Message-ID: <1ee.11186fa8.2cb621fe@aol.com> Check the temperature of the water on your stainer. Roxanne -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031008/3b039bac/attachment.htm From nick.kirk3 <@t> btopenworld.com Wed Oct 8 23:28:55 2003 From: nick.kirk3 <@t> btopenworld.com (Nick Kirk) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Purple Haze..... In-Reply-To: <1ee.11186fa8.2cb621fe@aol.com> Message-ID: Roxanne I can't see how the temperature of the water on the stainer could be causing this artefact as the problem tends to be intermittent even when batch staining. Sometimes within a rack of slides you will get some slides from GI biopsies showing the artefact while others from the same clinic session do not. It has to be something individual to each specimen in my opinion, otherwise we would see every slide going through on that particular batch exhibiting the same staining artefact, which we don't. I still think the actual biopsy process has a major part to play, especially as that's the only variable we don't have any control over and all the others we do. The temperature thing is a spurious argument in my opinion when you consider all blocks are chilled prior to cutting so everything is cold at some point and if you do immuno antigen retrieval you apply excessive heat to the slide and still get good nuclear detail when the section is counterstained. I think it has to be something else. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of GREYTRUNK@aol.com Sent: 09 October 2003 03:29 To: ryaskovich@dir.nidcr.nih.gov; STEGTM@samcstl.org; KMMerril@LancasterGeneral.org; histonet@pathology.swmed.edu Subject: Re: [Histonet] Purple Haze..... Check the temperature of the water on your stainer. Roxanne -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031009/ec29eb3b/attachment.htm From Myri37 <@t> aol.com Wed Oct 8 23:55:51 2003 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] MMA sections Message-ID: <3d.35f6c51c.2cb64457@aol.com> Skipped content of type multipart/alternative-------------- next part -------------- A non-text attachment was scrubbed... Name: not available Type: image/jpeg Size: 2082 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031009/d8e5526d/attachment.jpg From ctsblack <@t> capeheart.uct.ac.za Thu Oct 9 01:21:36 2003 From: ctsblack <@t> capeheart.uct.ac.za (Melanie Black) Date: Fri Sep 16 15:22:01 2005 Subject: Fwd: Re: [Histonet] MMA sections Message-ID: Hi Marseille This works very well, you need to de-resin in Xylene at 37 grees. Two changes for 15 minutes each. Then go on with normal wax staining process, i.e dehydrate, block primary etc, etc. You can find papers written by Neil Hand on this subject. It is tried and tested with great results. Good Luck Melanie Black Cardio Vascular Research University of Cape Town South Africa. >Date: Thu, 9 Oct 2003 08:16:20 +0200 >To: Myri37@aol.com >From: Melanie Black >Subject: Re: [Histonet] MMA sections >Cc: >Bcc: >X-Attachments: > >>Hello everyone >>i need to do ummunostaining on MMA sections of 5 Um deplasticised >>in xylene, i tried one time with a protocol used for paraffin >>section, but it doesn't work, i deplasiticised MMA in aceton 2 X 10 >>minutes at 56 ?C >>Please, do you have a protocol especially for MMA sections, or >>special embedding in MMA for immunostaning, or any advice for >>deplasticise.. >>thank you very much in advance >>Myriam Baali >> >>Natural Implant >>Marseille -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031009/5e762178/attachment.htm From binma2 <@t> web.de Thu Oct 9 03:29:21 2003 From: binma2 <@t> web.de (BIN MA) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] IHC staining problem with 30 um mouse lymph node frozen section Message-ID: <200310090829.h998TLQ18916@mailgate5.cinetic.de> An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031009/60470ee9/attachment.htm From L.D.Ridley <@t> bham.ac.uk Thu Oct 9 03:43:36 2003 From: L.D.Ridley <@t> bham.ac.uk (LD Ridley) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] SV40 staining Message-ID: <1065689016.9957c700L.D.Ridley@bham.ac.uk> Hi all, I'm still relatively new to immunohistochemistry but I seem to have hit a bit of a problem and would really appreciate any help. My problem is that I'm trying to stain SV40 T antigen in monkey kidney infected with SV40 (FFPE)using the Chemate Envision detection kit. I have had success with staining infected Cos7 cells using the antibody bought from Santa Cruz (Pab 101) sc-147 using extreme antigen retrieval techniques (so i know my antibody and technique seems fine). However when i try to stain the kidney slides i cant get any positivity. i've tried everything in vain. The guy that sent me the block can get nice staining using clone Pab416 My question really to anyone out there is.. can one antibody work on one tissue but not on another? and can different clones of an antibody work on one tissue and not on another even though they are both 'looking for' the same antigen.I have done a thorough literature search and both clones should detect the Large T antigen of SV40 so i'm reluctant to give up on Clone Pab 101 before rushing out to buy clone Pab 416. Sorry this quite a long one but any help from anyone working with SV40 would really be appreciated. I'll probably need to clear a few things up that i havn't explained very well here. Thanks in advance Lee Lee Ridley Research Associate L.D.Ridley@bham.ac.uk Birmingham Women's Hospital Reproductive & Child Health Academic Unit 3RD Floor Edgbaston Birmingham United Kingdom B15 2TG Tel: +44 (0)121 6074754 From Salvacion.DeLovino <@t> cshs.org Thu Oct 9 06:55:00 2003 From: Salvacion.DeLovino <@t> cshs.org (DeLovino, Salvacion S.) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Sentinel nodes Message-ID: After the first H&E cut, we cut another for IHC. Then we go 40 microns deeper then cut another set of H&E/IHC slides. Then we go another 40 microns deeper and cut the final set. So we have a total of 6 slides. That's if we have a big enough tissue. If not, then just adjust how deep we go depending on the size or thickness of the specimen. Salvacion Delovino -----Original Message----- From: JENNIFER SCHUMACHER [mailto:jschuma1@FAIRVIEW.ORG] Sent: Wednesday, October 08, 2003 1:35 PM To: histonet@lists.utsouthwestern.EDU Subject: [Histonet] Sentinel nodes Good afternoon everyone, We routinely cut our negative sentinel node biopsies shallow, cutting 5 levels (mounting levels 2 and 4 for IHC), approximately 30 microns between levels. Now, a pathologist read that "most institutions" are cutting them deeper, approximately 100 microns between levels. This is totally an INFORMAL poll, but I would appreciate any feedback from the community. I will check the archives as well, but thought I would get the question out there before everyone heads off to Kentucky:-) Thanks. Jennifer Important Warning: This message is intended for the use of the person or entity to which it is addressed and may contain information that is privileged and confidential, the disclosure of which is governed by applicable law. If the reader of this message is not the intended recipient, or the employee or agent responsible to deliver it is the intended recipient, you are hereby notified that any dissemination, distribution or copy of this information is STRICTLY PROHIBITED. If you have received this message by error, please notify us immediately by calling (310) 423-6428 and destroy the related message. Thank you for your cooperation. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031009/038a13c7/attachment.htm From cgfields <@t> lexhealth.org Thu Oct 9 08:16:09 2003 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] warthin starry Message-ID: Hi Celia, We also had problems with gastric bx's and it wasn't a consistent problem. I retrieved them and then stained them H&E and they were much better. It did not seem to matter what you retrieved them with either....very weird. We think it is something with the fixation. I had a problem with bone marrows and it turned out to be, the decal was messed up, but I retrieved them and they stained better. Go figure.... but it worked. My Pathologist also heard about this at a conference he went to. They did not know why it worked either. We do the microwave Steiner now and it works much better. Try the retrieval...good luck. Carole Fields Lexington Medical Center W.Columbia, SC > -----Original Message----- > From: celia [SMTP:cfarm@sio.midco.net] > Sent: Wednesday, October 08, 2003 6:23 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] warthin starry > > Hello Everyone, > > I have put this question out once before and I didn't get much for a > response so I thought I would try putting it out there one more time. I > have an ongoing problem with my Warthin-Starry stain. The peripheral > edges of some of the gastric biopsies are not staining. I am also > experiencing this same non-staining with the Steiner stain on the Ventana > stainer. I have tried reducing the temp on the processor down from 60c > to 58c thinking they might be "cooking" on the processor (the H&E's look > fine) as well as checking the ph of my solutions. This did not work. > Could this be a handling problem prior to fixation in 10% NBF? I am at > my wits end. Any suggestions will be greatly appreciated. > > Thanks in advance. > > Celia, HT > SVH, Sioux Falls, SD > > > _________________________________ 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. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031009/1e0092c7/attachment.htm From Ronnie_Houston <@t> bshsi.com Thu Oct 9 08:29:04 2003 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Association for Molecular Pathology Message-ID: <530361BF03351B4CAE5270A05D3037B5FE0481@bsrexms01.BSHSIR.COM> Mohammed, Check out the association's web-site http://www.ampweb.org/ Ronnie -----Original Message----- From: Sayeed, Mohammed [mailto:Sayeed@www.urol.bcm.tmc.edu] Sent: Thursday, October 09, 2003 9:13 AM To: 'Houston, Ronnie' Subject: RE: [Histonet] Association for Molecular Pathology Dear Ronnie Can you provide me and the other Histonet readers more info about the meeting in Orlando. I would like to attend. M. Sayeed Cheif Tech Spore Pathology Baylor college of Medicine. > -----Original Message----- > From: Houston, Ronnie [SMTP:Ronnie_Houston@bshsi.com] > Sent: Wednesday, October 08, 2003 11:33 AM > To: Histonet (E-mail) > Subject: [Histonet] Association for Molecular Pathology > > > Anyone going to the annual meeting of the Association of Molecular > Pathology > meeting in Orlando, Nov 20-23, 2003? > > Ronnie Houston > Regional Histology Operations Manager > Bon Secours HealthPartners Laboratories > 5801 Bremo Road > Richmond, VA 23226 > (804) 287 7972 > 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. > > > _______________________________________________ > 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 djcarter <@t> dallastx.net Thu Oct 9 08:30:19 2003 From: djcarter <@t> dallastx.net (Dallas Nippert) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Purple Haze..... References: Message-ID: <001901c38e69$825318a0$1bfcf3d0@djcarter> If you use a slide adhesive such as sta-on in your waterbath it will cause a purple haze on the slide if you use too much. Dallas ----- Original Message ----- From: Nick Kirk To: Histonet Sent: Wednesday, October 08, 2003 11:28 PM Subject: RE: [Histonet] Purple Haze..... Roxanne I can't see how the temperature of the water on the stainer could be causing this artefact as the problem tends to be intermittent even when batch staining. Sometimes within a rack of slides you will get some slides from GI biopsies showing the artefact while others from the same clinic session do not. It has to be something individual to each specimen in my opinion, otherwise we would see every slide going through on that particular batch exhibiting the same staining artefact, which we don't. I still think the actual biopsy process has a major part to play, especially as that's the only variable we don't have any control over and all the others we do. The temperature thing is a spurious argument in my opinion when you consider all blocks are chilled prior to cutting so everything is cold at some point and if you do immuno antigen retrieval you apply excessive heat to the slide and still get good nuclear detail when the section is counterstained. I think it has to be something else. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of GREYTRUNK@aol.com Sent: 09 October 2003 03:29 To: ryaskovich@dir.nidcr.nih.gov; STEGTM@samcstl.org; KMMerril@LancasterGeneral.org; histonet@pathology.swmed.edu Subject: Re: [Histonet] Purple Haze..... Check the temperature of the water on your stainer. Roxanne -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031009/6ab4f324/attachment.htm From docmichel <@t> netbulmail.com Thu Oct 9 08:42:08 2003 From: docmichel <@t> netbulmail.com (izzet oguz) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] (no subject) Message-ID: <2358.193.255.128.130.1065706928.webmail@mail1.netbulmail.com> Hi Histoworld, Is there any spesific marker for angioneogenesis or immature blood vessels? Thanks, ?zzet From AFeatherstone <@t> KaleidaHealth.Org Thu Oct 9 08:56:02 2003 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Shattering Mouse Brain Message-ID: Sounds like processing to me. Times probably need to be changed and solutions need to be fresh. Annette HT/MLT -----Original Message----- From: LaTricia Faison [mailto:lfaison@mail.mcg.edu] Sent: Wednesday, October 08, 2003 12:27 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Shattering Mouse Brain Does anyone know what may cause mouse brain to separate when put on the water bath? The tissue was given to me after it had been in paraformaldehyde for about 7-8 months. It was paraffin processed and shattered soon after touching the water bath. _______________________________________________ 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 HornHV <@t> archildrens.org Thu Oct 9 09:03:58 2003 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] negative IHC controls Message-ID: Not only should you be running a negative control for each patient slide. That negative control should be treated just as your antibody is. If the antibody is rabbit and antigen retrieved, so should your control. If another antibody on the same patient is mouse and not retrieved another negative control should be run with this same protocol. In the United States, labs that are inspected by the CAP, are required to run these controls. MONEY should never be considered as a reason to stop doing a part of a procedure. It's poor patient care and lousy quality control. IMHO. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: vermast [mailto:vermast@rogers.com] Sent: Wednesday, October 08, 2003 3:57 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] negative IHC controls I would like to get a feel for how many out there are running negative control slides for IHC. In our lab we do just a handful of antibodies and initially I had been running a negative control slide with each patient slide. After much discussion with our pathologists, we decided to omit these negatives (which were conistently negative) and continue to just run a positive control with each primary antibody for the run. We use the Dako autostainer and prediluted primaries. The decision to stop running negatives also coincided with Dako's decision to sell the negative control sera separately from the primaries (they used to come packaged together). Perhaps I assumed that discontinuing to pair these reagents together meant that few labs were using the negatives. Anyhow, after having reviewed the last QMPLS (Canada) survey committee comments, I believe the committe would like a negative control run with each patient tissue slide in order to evaluate background (they have used NCCLS guide pages as reference). Incidentally we weren't a part of the survey due to a technicality. Any help or advice would be appreciated. L. Vermast Stratford, Ont. 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. Arkansas Children's Hospital -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031009/6d9935bd/attachment.htm From nick.kirk3 <@t> btopenworld.com Thu Oct 9 09:23:42 2003 From: nick.kirk3 <@t> btopenworld.com (nick.kirk3@btopenworld.com) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Purple Haze..... Message-ID: <2383268.1065709422277.JavaMail.root@127.0.0.1> No slide adhesive used, this appears on standard non-charged, non-adhesive coated slides. This artefact is localised to the cell nuclei only. I know what you mean though, I remember in the old days when we used to albuminise slides, they always had a blue wash over them. Nick > from: Dallas Nippert > date: Thu, 09 Oct 2003 14:29:09 > to: nick.kirk3@btopenworld.com > subject: Re: [Histonet] Purple Haze..... > > If you use stay-on or another slide adhesive in your water bath, you may be using too much. This will cause the hematoxylin to stain the slide. > Dallas > ----- Original Message ----- > From: Nick Kirk > To: Histonet > Sent: Wednesday, October 08, 2003 11:28 PM > Subject: RE: [Histonet] Purple Haze..... > > > Roxanne > > I can't see how the temperature of the water on the stainer could be causing this artefact as the problem tends to be intermittent even when batch staining. > Sometimes within a rack of slides you will get some slides from GI biopsies showing the artefact while others from the same clinic session do not. > It has to be something individual to each specimen in my opinion, otherwise we would see every slide going through on that particular batch exhibiting the same staining artefact, which we don't. > I still think the actual biopsy process has a major part to play, especially as that's the only variable we don't have any control over and all the others we do. > The temperature thing is a spurious argument in my opinion when you consider all blocks are chilled prior to cutting so everything is cold at some point and if you do immuno antigen retrieval you apply excessive heat to the slide and still get good nuclear detail when the section is counterstained. > I think it has to be something else. > > Nick Kirk > Histopathology > Hinchingbrooke Hospital > Huntingdon > England > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of GREYTRUNK@aol.com > Sent: 09 October 2003 03:29 > To: ryaskovich@dir.nidcr.nih.gov; STEGTM@samcstl.org; KMMerril@LancasterGeneral.org; histonet@pathology.swmed.edu > Subject: Re: [Histonet] Purple Haze..... > > > Check the temperature of the water on your stainer. > Roxanne From ccrowder <@t> mail.vetmed.lsu.edu Thu Oct 9 10:02:49 2003 From: ccrowder <@t> mail.vetmed.lsu.edu (Cheryl Crowder) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Mouse brain Message-ID: Annette - We have worked with a lot of mice brains and have had your problem. We cut all the brains and CNS tissues at one time (not interspersed with other pieces and parts). We use a waterbath at 37C. This helps the paraffin stay next to the tissue. One of the problems with mice is that they can over-dehydrate easily and need shorter processing times. When the tissue hits the water it "goes crazy" sucking up the water into the dehydrated tissues. We also have moistened a slide and then placed the section directly on it, draining the excess water. Try that if all else fails. Hope this helps. Cheryl Cheryl Crowder, BA, HTL(ASCP) Chief Technologist Anatomic Pathology Department of Pathobiological Sciences School of Veterinary Medicine Louisiana State University Skip Bertman Drive Baton Rouge, LA ?70803 225-578-9734 FAX: ?225-578-9720 From nancymaronto <@t> sbcglobal.net Thu Oct 9 10:04:58 2003 From: nancymaronto <@t> sbcglobal.net (Nancy Maronto) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] Need silver stain for granules in adrenals Message-ID: <20031009150458.75311.qmail@web80109.mail.yahoo.com> Hi, Could anyone suggest a stain for noradrenalin and adrenalin granules in paraffin embedded adrenals. We were asked to do a gremilius stain and it did not stain the granules. Are these slides lost or can we re stain them? We usually do a recut of the block, but it is asked if we could use the same tissue slide. The stain actually worked, but not for the granules. >From what we read a Fontana-masson should work. There probably is a better stain our there for this specific target. Can anyone with some experience with staining these granules share their staining protocol. At this time the request is not for immuno staining. Thanks for your help. Nancy Maronto -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031009/1a5f172d/attachment.htm From msandova <@t> nd.edu Thu Oct 9 10:06:40 2003 From: msandova <@t> nd.edu (msandova@nd.edu) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] IHC staining problem with 30 um mouse lymph node frozen section In-Reply-To: <200310090829.h998TLQ18916@mailgate5.cinetic.de> References: <200310090829.h998TLQ18916@mailgate5.cinetic.de> Message-ID: <1065712000.3f8579803352f@webmail.nd.edu> I'm curious, why are the sections 30um thick? Mayra Quoting BIN MA : > p {margin: 0px}Dear All, I have a problem with IHC staining(using > Perioxdase,AEC substrate)) on 30 um thick mouse lymph node frozen sections. > After staining part of section is washed away and the morphology of the > section is poor. Can someone help me? My protocol is following: > IHC staining protocol Cut 30 um frozen > sections.Fix in cold methanol for 3 minutes, then in cold aceton for 3 > minutes.Air dry for 1 hour.Put the section in PBS for 10 minutes.Block with > 1%FCS/PBS 100 ul for 20 minutes at room temperature.Endogenous biotin > blocking:(1) Incubate slice in egg solution for 15 minutes.Egg solution: 25 > ml egg white ,75 ml distilled water.Wash with 2 changes of distilled > water.(2) Incubate slice in the milk(fat free) for 15 minutes at room > temperature. Wash with 2 changes of distilled water.Add diluted first > antibody.For example :CD3-biotin Incubate 2 hours at room temperature or at 4 > ?C overnight.Wash in PBS for 30 seconds, 3 minutes and 10 minutes three > times.endogenous peroxidase blocking:The solution : 0,3% H2O2 ,0,25% NaN3 > in PBS Incubate the section in this solution for half hour. Rinse with water > for 2 minutes. Wash in PBS for 5 minutes.8 Block with 1%FCS/PBS for > another 20 minutes at room temperature.9. Add second antibody for example > streptavidin-PO Incubate for 4 hours at 4 ?C.. Wash with PBS > for 1 ,3 ,10 minutes for three times.9 Add substrate 300 ul for each > section, incubate at 37?C surface for half hour.Substrate solution: 25 ul > AEC stock solution 1 ml Na-Acetate > buffer(5,4) 1ul H2O2Attention: Make this > solution just before use,protect it from light..Wash in PBS for 30 seconds,3 > minutes and 10 minutes for three times.10 Dry on 37?C surface for 5 > minutes.11 Mount with 175 ul Kaser?s galatine using 24x60 cover > glass. 12 Read the slides. Will adding Triton or tween 20 help ? Thank > you.Bin maGerman National research Centre > ofBiotechnologyBraunschweigGermany > ______________________________________________________________________________ > Die Besten ihrer Klasse! WEB.DE FreeMail (1,7) und WEB.DE Club (1,9) - > bei der Stiftung Warentest - ein Doppelsieg! http://f.web.de/?mc=021184 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From histo <@t> bthosp.com Thu Oct 9 10:03:02 2003 From: histo <@t> bthosp.com (O'Brien, Sue) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] SNO-Med and ICD-9 Codes Message-ID: <3653750027C5D6119D0600508BB89A9A0ABF4D@MAIL_SERVER> I was just wondering if there were any reason why SNO-Med terminology should be updated in our computer data base? Now that we have ICD-9 codes, is the SNO-Med terminology necessary? for example, for any regulatory agency requirement? Just wanted some input on this, Thanks, Sue O'Brien, Histology Supervisor Burdette Tomlin Memorial Hospital Cape May Court House, NJ 08210 From ckbyrne <@t> exelixis.com Thu Oct 9 10:06:57 2003 From: ckbyrne <@t> exelixis.com (Carrie Kyle-Byrne) Date: Fri Sep 16 15:22:01 2005 Subject: [Histonet] SV40 staining References: <1065689016.9957c700L.D.Ridley@bham.ac.uk> Message-ID: <000401c38e76$fc1b7990$860c1dac@ckbyrnewkst> Hi Lee, i'd say that the problem you are experiencing is related to the fixation of the tissue your collaborator has sent you. my suggestion would be, and you may have already done this, to play with the length of time spent in the antigen retrieval.....perhaps going even longer that what you normally use for the Cos7 cells (they were fixed overnight in 10% NBF). Pab101 should work for you just fine reguardless of species ....it's seeing a virus protein.....not a protein that is species specific. (for those not familiar, Cos7 is a line derived from monkey kidney). Good luck, Carrie Kyle-Byrne, BHS, HT(ASCP) Assoc. Research Scientist II Antibody Technology Lab Signal Transduction Research Exelixis, Inc. 170 Harbor Way P.O. Box 511 South San Francisco CA 94083-0511 USA Phone: (1 650) 837-8023 Fax: (1 650) 837-7220 Email: ckbyrne@exelixis.com FedEx Deliveries and Shipping to: 169 Harbor Way South San Francisco CA 94080 USA ________________________________________________________________ This email message 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 email and destroy all copies of the original message. ----- Original Message ----- From: "LD Ridley" To: Sent: Thursday, October 09, 2003 1:43 AM Subject: [Histonet] SV40 staining > Hi all, > I'm still relatively new to immunohistochemistry but I seem to have hit a bit of a problem and would really appreciate any help. My problem is that I'm trying to stain SV40 T antigen in monkey kidney infected with SV40 (FFPE)using the Chemate Envision detection kit. I have had success with staining infected Cos7 cells using the antibody bought from Santa Cruz (Pab 101) sc-147 using extreme antigen retrieval techniques (so i know my antibody and technique seems fine). However when i try to stain the kidney slides i cant get any positivity. i've tried everything in vain. The guy that sent me the block can get nice staining using clone Pab416 My question really to anyone out there is.. can one antibody work on one tissue but not on another? and can different clones of an antibody work on one tissue and not on another even though they are both 'looking for' the same antigen.I have done a thorough literature search and both clones should detect the Large T antigen of SV40 so i'm reluctant to give up on Clone Pab 101 before rushing out to buy clone Pab 416. > Sorry this quite a long one but any help from anyone working with SV40 would really be appreciated. I'll probably need to clear a few things up that i havn't explained very well here. > Thanks in advance > Lee > Lee Ridley > Research Associate > > L.D.Ridley@bham.ac.uk > > Birmingham Women's Hospital > Reproductive & Child Health > Academic Unit 3RD Floor > Edgbaston > Birmingham > United Kingdom > B15 2TG > > Tel: +44 (0)121 6074754 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From mcauliff <@t> umdnj.edu Thu Oct 9 10:53:11 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Perfusion debate In-Reply-To: References: Message-ID: <3F858467.70507@umdnj.edu> Continuing .......... Charles W. Scouten, Ph.D. wrote: >The large diameter needle into the ventricle sounds great. > Works for me. >High flow rate sounds good to get clean of red blood cells, but how high is high, and in what species and body size? > Good question, I never did any real calculations. Let's see, blood volume is about 8% of body weight in man so I will stick with that number unless someone has a better (documented) one. I will also assume that cardiac output is proportional in man/mouse/rat. Man is 5 liters/minute and the circulating blood volume is about 5 liters. For a 30 gram mouse (30x8%), about 2.4 ml of blood circulating every minute.When I perfuse a mouse I pump in about 60-75 ml in 5 minutes, lots more than the circulating volume over the same amount of time. For a 300 gram rat (about 24 ml of circulating blood) I pump in a minimum of 300 ml in 5 minutes, again much more than would circulate over the same amount of time. I rarely have a perfusion "fail" and I don't have problems with shrinkage, distorted cells, tissues, etc. I suspect that the high volumes I perfuse in a short amount of time are the cause. I have never measured the pressure my pump puts out with the needle sizes I use, perhapsI should find a gauge of some sort. >Of course, pressure and flow rate are closely related, > Yes. > the third determining variable is cardiovascular resistance, which is related to size, gender, species, health condition, etc. A mouse would have far more resistance to a given flow than would a pig. > I don't think so. One can have very high flow and very low pressure. A large diameter hose will move a lot of fluid with very low pressure. Cardiovascular dynamcs (see your table below) are reasonably similar in mammals, given good health. Gender differences would be small, but measurable. Resistance is primarliy determined by the (cumlative) cross-sectional area of the vessels, hence the pressure drop in the arterioles just before to the capillary beds. Any fluid, not just blood, will behave this way. It's Poiseuill's Law. > However, as might be guessed, all mammals have very close to the same average blood pressure. See the table below, (pointed out to me by histonetters, thank you again): >The following is a table of average blood pressure for several species (systolic/diastolic) (Green, 1979). > >mice 113/81 rats 116/90 hamster 150/110 >rabbit 110/80 dog 112/56 cat 120/75 >baboon 148/100 rhesus 160/127 pig 170/108. > >Consider what would happen if you perfused a pig with the same flow rate that works well on a mouse, or vice versa. But you can use the same pressure to perfuse a pig or mouse, and get comparable results. > > Controlling pressure is safer and more predictable. > >A high flow rate means high pressure, > No! Related yes, but not that way. To get one liter of solution through a 16 gauge needle in one minute will require a lot less pressure than moving the same amount of fluid through a 25 gauge needle in the same amount of time. >but the operational definition of high flow rate must depend on the species and body size, among other variables. High but safe pressure is 200 mm Hg in all of the above. The blood brain barrier is reliable broken in hamsters and rats at 300 mm Hg. What do you bet for the rest? > > >At 300 mm Hg, I come very close to washing out every last RBC in Hamster or Rat brain. HRP shows maybe one in each whole brain coronal 60 micron thick section. There is no noticeable (by me) damage to capillaries, but maybe a better anatomist could see it. > >You do have shrinkage in the CNS, it is well documented. > References, please. So when I remove the skull from an animal perfused my way why isn't the brain 20% smaller than a fresh brain? > You don't see it as a problem because it has always been there, and you haven't worked with fixed sections without it. See Konig and Klippel for "inevitable shrinkage", and ask Paxinos why his atlases are done only on fresh, not fixed tissue. But you can have both fixed and no shrinkage. You EM sections showed little or no extracellular space. > My EM sections show an amount of extracellular space appropriate for the tissue. Not much in CNS, lots in the dermis. >Actually, the 5% sucrose is slightly hypertonic, isotonic is something like 4.94 by weight. I had never heard that point, though, can you tell me more about the documentation on that? > Arborgh, et al, J. Ultrastructure Research 56:339-350, 1976. Not perfusion fixation, but verifying that the osmolality of the buffer the fixative is in, not the osmolality of fixative, is important. There is lots of work on fixation and shrinkage, time in fixative, pH of fixative, etc. done in the 1950's and 1960's. Both model systems and tissues were used. > Kidney is different, there is no blood brain barrier, and near isotonic sucrose would replace the extracellular fluid without the high pressure. > The blood-brain barrier is not a barrier to pressure, it is much more of a diffusion/metabolic barrier. > Isn't the active sodium pump limited to only exciteable cells? > No, all cells have an ATP-driven sodium-potassium pump to keep Na outside and K inside. > If so, this formalin shrinkage thing would only be relevant for neural or muscle cells. Does anybody know if kidney or liver, for example, shrink with perfusion? > > > 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 >www.myneurolab.com > > Gotta get ready for class. Geoff > >-----Original Message----- >From: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] >Sent: Wednesday, October 08, 2003 1:12 PM >To: histonet@pathology.swmed.edu >Subject: Re: [Histonet] Perfusion debate > > Joining in the perfusion debate......... I have perfused mouse and >rat CNS with a pump and had excellent results. I have never measured the >pressure the pump puts out but I do use a syringe needle (into the left >ventricle) with a large diameter, 16 gauge for a rat, 20 gauge for a >mouse. I think something close to the inside diameter of the aorta is >required for good results. In my experience, high rate of flow is more >important that pressure. I don't worry about washing out every last RBC, >I don't think it can be done. I use a minimum amount of buffer (a few >milleliters) as a prewash and I don't use vasodilators or >anti-coagulents. No shrinkage problems for LM or EM. I have never tried >sucrose as a prewash. > I have also perfused rat kidney with just two jars high on a shelf >above the sink. Beautiful EM's from that work, even from the renal >medulla which has relatively low blood flow. Again, I did not measure >the pressure but it was only about 90-100 mm Hg (converting the height >in inches to mm then converting from water to mercury using specific >gravity). > To summarize, use a high rate of flow and be sure the buffer (not the >fix+buffer solution) you use is somwhat hypertonic. This last point is >well documented in the literature. > >Geoff > >mucram11@earthlink.net wrote: > > > >>Strange we did not have shrinkage problems with our set up. Pam Marcum >> >>Original Message: >>----------------- >>From: Charles W. Scouten, Ph.D. cwscouten@myneurolab.com >>Date: Wed, 8 Oct 2003 10:39:33 -0500 >>To: mucram11@earthlink.net, frouwke@sci.kun.nl, >>nancy.walker@sanofi-synthelabo.com, histonet@lists.utsouthwestern.edu >>Subject: RE: [Histonet] RE: shrinkage and Perfusion One system >> >> >>Yes, gravity will work, with 20% shrinkage of the brain, some shape >>distortion, and with a reddish tinge due to remaining red blood cells, >>which catalyze any HRP reaction, and autofluoresce. OK for some >>purposes, >>terrible for others, but not the optimum for any. >>I have asked www.Histonet.org to post a picture called Perfused Brains >>Annotated which shows whole rat brains, one fresh dissected out, one >>perfused by pressure sucrose and 4% paraformaldehyde/glut, and the >>last by >>traditional gravity flow with saline prewash, and the same fixative. >> >>The last is reddish and shrunken compared to the middle brain. >>Three ft of water translates directly, using the units converter at: >>http://www.sciencemadesimple.com/conversions.html >>to 67 mm Hg, below average diastolic pressure in mice or rats or >>people. And some pressure is lost in the lines and needle. Even in >>living animals >>with cycles of systolic pressure, some capillaries occasionally get >>plugged >>for a while and stop flowing. That is part of the advantage of exercise, >>to keep the lines open. It is extremely unlikely that completely open >>exposure to 67 mm Hg would in any way "blow" the vascular system of any >>mammal, or even clear out all the blood. The blocked passages will stop >>fixative from reaching the surrounding tissue in that area, and lead to >>local deterioration in unpredictable and irreproduceible areas. >> >>The only advantage of sucrose over saline is to clear the extracellular >>fluid of sodium before fixative hits. High pressure, 300 mm Hg, >>applied to >>either saline or sucrose as the prewash, (there is never a reason for >>both), will thoroughly clear all red blood cells, but should not be >>pumped >>up in advance the 'thump' the system, but increase over a few seconds to >>clear most blood before the blood brain barrier opens. This prewash will >>take half the time of traditional prewash, and get fixative in sooner and >>more thoroughly. >> >>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 www.myneurolab.com >> >>-----Original Message----- >>From: mucram11@earthlink.net [mailto:mucram11@earthlink.net] Sent: >>Wednesday, October 08, 2003 8:06 AM >>To: frouwke@sci.kun.nl; Charles W. Scouten, Ph.D.; >>nancy.walker@sanofi-synthelabo.com; histonet@lists.utsouthwestern.edu >>Subject: Re: [Histonet] RE: shrinkage and Perfusion One system >> >>Hi, >>We never had a pump. We used gravity for over 2,000 rats so I think I can >>say it will work. The height of the container or IV bottle used >>seemed to >>be the key. We controlled flow with the standard IV roller closure on >>the >>tubing. It was approximately 3ft above the area the animal was being >>perfused in, always under a hood or very well ventilated area. We looked >>for a steady drip in the tubing and out the cannula. We did not open the >>valve all the way as it would blow out the vascular system. We did have a >>couple of students try wide open and prove the point very well. >>The use of a saline solution with and without anticoag agents had been >>used. If you are going to use sucrose I would use the pre-wash to get >>the >>blood and red cells out. Just to be sure you have all the small vessels >>cleared for the fixative or freezing technique. >>Pam Marcum >> >>Original Message: >>----------------- >>From: Frouwke Kuijpers frouwke@sci.kun.nl >>Date: Wed, 8 Oct 2003 12:47:14 +0200 >>To: cwscouten@myneurolab.com, Nancy.Walker@sanofi-synthelabo.com, >>Histonet@lists.utsouthwestern.edu >>Subject: Re: [Histonet] RE: shrinkage and Perfusion One system >> >> >>Start the prewash at low pressure, and pump up to 300 mmHg >>to break the blood brain barrier and wash the extracellular fluid (and >>red >>blood cells). >> >>Charles: next question: we don't do the perfusion by a pump, just by >>gravity. You said the pressure for the 10% sucrose must go till >>mm300Hg.... >>Well we dont' have a pump: what will your advice be than? >>Start with the saline solution to get rid of the blood and than the >>sucrose? >> >>Frouwke Kuijpers >> >>----- Original Message ----- From: "Charles W. Scouten, Ph.D." >> >>To: ; >> >>Sent: Monday, October 06, 2003 3:53 PM >>Subject: [Histonet] RE: shrinkage and Perfusion One system >> >> >>The saline prewash would be just lost time. It accomplishes nothing that >>will not be accomplished by the sucrose, which is equally effective at >>washing out the blood (more effective given the high pressure). Start >>with >>the sucrose. Time is of the essence, tissue deterioration begins >>immediately upon anoxia. Switch to fixative when the animal's muscles >>cease >>to move. >> >>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 >>www.myneurolab.com >> >> >>-----Original Message----- >>From: Nancy.Walker@sanofi-synthelabo.com >>[mailto:Nancy.Walker@sanofi-synthelabo.com] >>Sent: Friday, October 03, 2003 8:12 AM >>To: Charles W. Scouten, Ph.D.; Histonet@lists.utsouthwestern.edu >>Subject: shrinkage and Perfusion One system >> >> >>Hello, >> >> >> >>>From the website mentioned in C. Scouten's mail I recuperated this info: >>> >>> >>The shrinkage occurs when a prewash with physiological saline is followed >>by fixative. The first action of fixative on the cell membrane is to shut >>off the sodium pump proteins. Sodium rushes into the cell, followed by >>water to maintain tonicity, and cells swell and expand. Membrane proteins >>are fixed and crosslinked to neighboring cells in the swollen position. >>Later, the cells stabilize and contract, and pull other cells with them. >>The result is gross shrinkage, local distortion and torn membranes with >>lost cellular contents from some cells. >> >>Tissue Shrinkage can be completely prevented if the extracellular fluid >>with ions is replaced by a nonionic isotonic solution that cannot >>enter the >>cells. This can be accomplished by prewash with 5% (isotonic) sucrose in >>distilled water. Start the prewash at low pressure, and pump up to 300 >>mmHg >>to break the blood brain barrier and wash the extracellular fluid (and >>red >>blood cells). Switch shortly thereafter to fixative. The perfusion takes >>the same time as the traditional procedure, and accomplishes the >>favorable >>results bulleted above. >> >>So if I'd like to try out the perfusion techniques that are made >>simple by >>the Perfusion one system, what would you suggest: first do a >>physiological saline solution, followed by a prewash with 5% sucrose and >>then PFA, or go directly to 5% sucrose then PFA? >> >>thanks for your advice, >> >>Nancy >> >> >> >> > > > -- -- ********************************************** 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 ********************************************** -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031009/a8b2625d/attachment.htm From STEGTM <@t> samcstl.org Thu Oct 9 11:01:38 2003 From: STEGTM <@t> samcstl.org (Therersa Stegall) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Purple Haze..... Message-ID: Problem solved! After considering amounts of Sta-on, gelatin, and using a "naked" waterbath, all still produced the artifact. It had to be the hematoxylin, eh? We used modified Harris, and considering the recipe has acetic acid in it, thought that this was not the optimal for decolorization in regressive staining. Thought the haze could be an indication of decaying hematoxylin; used HCl in our acid/water solution this am, maybe it would help to capture the possibe alum hematein.... anyhow, it worked. The slides are crisp and clear, free from artifact, and now that the pathologists are happy, we are.......Thanks, Terre >>> "Yaskovich, Ruth A (NIH/NIDCR)" 10/08/03 01:48PM >>> I used to get this problem once in awhile and haven't had it since switching to charged slides. May-be in the amounts of gelatin? What kind of hematoxylin are you using? Ruth Yaskovich N.I.D.C.R. N.I.H. -----Original Message----- From: Therersa Stegall [mailto:STEGTM@samcstl.org] Sent: Wednesday, October 08, 2003 1:50 PM To: KMMerril@LancasterGeneral.org; histonet@pathology.swmed.edu Subject: [Histonet] Purple Haze..... Kathleen: Thanks for naming the amorphous blight that has been plaguing us lately. This "Jimi Hendrix" artifact has been running 'round our brains for the past few weeks. We have 1-started filtering the hematoxylin each morning, 2-alternated between Sta-on, gelatin, and naked waterbaths, 3-scrubbed the entire staining system, 4-decreased time in microwave, ovens, for drying slides; and yet, we still find this on some biopsies. Not always all the biopsies in a batch, this led us to ask GI about collection procedures, and possible air-drying, they insist it's not them. We've checked pH on water, accurately calibrated the bluing and acid/water washes (we stain regressively), and just about run out of ideas. I'm beginning to suspect the hematoxylin is leaving a precipitate, or maybe the microtomy is inconsistent and ruining cell integrity, or .... maybe it's terrorism! Any ideas out there? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Thu Oct 9 11:08:19 2003 From: cwscouten <@t> myneurolab.com (Charles W. Scouten, Ph.D.) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Perfusion debate Message-ID: For those following the perfusion debate, the pictures mentioned are now posted on Histonet.org, click "View Listserve Images" and scroll to the bottom for the most recent postings. Click "Perfused Brains, annotated copy" A picture of 3 rat brains; fresh, perfused with pressure sucrose method, and perfused with gravity flow. 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 www.myneurolab.com -----Original Message----- From: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] Sent: Wednesday, October 08, 2003 1:12 PM To: histonet@pathology.swmed.edu Subject: Re: [Histonet] Perfusion debate Joining in the perfusion debate......... I have perfused mouse and rat CNS with a pump and had excellent results. I have never measured the pressure the pump puts out but I do use a syringe needle (into the left ventricle) with a large diameter, 16 gauge for a rat, 20 gauge for a mouse. I think something close to the inside diameter of the aorta is required for good results. In my experience, high rate of flow is more important that pressure. I don't worry about washing out every last RBC, I don't think it can be done. I use a minimum amount of buffer (a few milleliters) as a prewash and I don't use vasodilators or anti-coagulents. No shrinkage problems for LM or EM. I have never tried sucrose as a prewash. I have also perfused rat kidney with just two jars high on a shelf above the sink. Beautiful EM's from that work, even from the renal medulla which has relatively low blood flow. Again, I did not measure the pressure but it was only about 90-100 mm Hg (converting the height in inches to mm then converting from water to mercury using specific gravity). To summarize, use a high rate of flow and be sure the buffer (not the fix+buffer solution) you use is somwhat hypertonic. This last point is well documented in the literature. Geoff mucram11@earthlink.net wrote: > Strange we did not have shrinkage problems with our set up. Pam Marcum > > Original Message: > ----------------- > From: Charles W. Scouten, Ph.D. cwscouten@myneurolab.com > Date: Wed, 8 Oct 2003 10:39:33 -0500 > To: mucram11@earthlink.net, frouwke@sci.kun.nl, > nancy.walker@sanofi-synthelabo.com, histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] RE: shrinkage and Perfusion One system > > > Yes, gravity will work, with 20% shrinkage of the brain, some shape > distortion, and with a reddish tinge due to remaining red blood cells, > which catalyze any HRP reaction, and autofluoresce. OK for some > purposes, > terrible for others, but not the optimum for any. > I have asked www.Histonet.org to post a picture called Perfused Brains > Annotated which shows whole rat brains, one fresh dissected out, one > perfused by pressure sucrose and 4% paraformaldehyde/glut, and the > last by > traditional gravity flow with saline prewash, and the same fixative. > > The last is reddish and shrunken compared to the middle brain. > Three ft of water translates directly, using the units converter at: > http://www.sciencemadesimple.com/conversions.html > to 67 mm Hg, below average diastolic pressure in mice or rats or > people. And some pressure is lost in the lines and needle. Even in > living animals > with cycles of systolic pressure, some capillaries occasionally get > plugged > for a while and stop flowing. That is part of the advantage of exercise, > to keep the lines open. It is extremely unlikely that completely open > exposure to 67 mm Hg would in any way "blow" the vascular system of any > mammal, or even clear out all the blood. The blocked passages will stop > fixative from reaching the surrounding tissue in that area, and lead to > local deterioration in unpredictable and irreproduceible areas. > > The only advantage of sucrose over saline is to clear the extracellular > fluid of sodium before fixative hits. High pressure, 300 mm Hg, > applied to > either saline or sucrose as the prewash, (there is never a reason for > both), will thoroughly clear all red blood cells, but should not be > pumped > up in advance the 'thump' the system, but increase over a few seconds to > clear most blood before the blood brain barrier opens. This prewash will > take half the time of traditional prewash, and get fixative in sooner and > more thoroughly. > > 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 www.myneurolab.com > > -----Original Message----- > From: mucram11@earthlink.net [mailto:mucram11@earthlink.net] Sent: > Wednesday, October 08, 2003 8:06 AM > To: frouwke@sci.kun.nl; Charles W. Scouten, Ph.D.; > nancy.walker@sanofi-synthelabo.com; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] RE: shrinkage and Perfusion One system > > Hi, > We never had a pump. We used gravity for over 2,000 rats so I think I can > say it will work. The height of the container or IV bottle used > seemed to > be the key. We controlled flow with the standard IV roller closure on > the > tubing. It was approximately 3ft above the area the animal was being > perfused in, always under a hood or very well ventilated area. We looked > for a steady drip in the tubing and out the cannula. We did not open the > valve all the way as it would blow out the vascular system. We did have a > couple of students try wide open and prove the point very well. > The use of a saline solution with and without anticoag agents had been > used. If you are going to use sucrose I would use the pre-wash to get > the > blood and red cells out. Just to be sure you have all the small vessels > cleared for the fixative or freezing technique. > Pam Marcum > > Original Message: > ----------------- > From: Frouwke Kuijpers frouwke@sci.kun.nl > Date: Wed, 8 Oct 2003 12:47:14 +0200 > To: cwscouten@myneurolab.com, Nancy.Walker@sanofi-synthelabo.com, > Histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] RE: shrinkage and Perfusion One system > > > Start the prewash at low pressure, and pump up to 300 mmHg > to break the blood brain barrier and wash the extracellular fluid (and > red > blood cells). > > Charles: next question: we don't do the perfusion by a pump, just by > gravity. You said the pressure for the 10% sucrose must go till > mm300Hg.... > Well we dont' have a pump: what will your advice be than? > Start with the saline solution to get rid of the blood and than the > sucrose? > > Frouwke Kuijpers > > ----- Original Message ----- From: "Charles W. Scouten, Ph.D." > > To: ; > > Sent: Monday, October 06, 2003 3:53 PM > Subject: [Histonet] RE: shrinkage and Perfusion One system > > > The saline prewash would be just lost time. It accomplishes nothing that > will not be accomplished by the sucrose, which is equally effective at > washing out the blood (more effective given the high pressure). Start > with > the sucrose. Time is of the essence, tissue deterioration begins > immediately upon anoxia. Switch to fixative when the animal's muscles > cease > to move. > > 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 > www.myneurolab.com > > > -----Original Message----- > From: Nancy.Walker@sanofi-synthelabo.com > [mailto:Nancy.Walker@sanofi-synthelabo.com] > Sent: Friday, October 03, 2003 8:12 AM > To: Charles W. Scouten, Ph.D.; Histonet@lists.utsouthwestern.edu > Subject: shrinkage and Perfusion One system > > > Hello, > >> From the website mentioned in C. Scouten's mail I recuperated this info: > > > The shrinkage occurs when a prewash with physiological saline is followed > by fixative. The first action of fixative on the cell membrane is to shut > off the sodium pump proteins. Sodium rushes into the cell, followed by > water to maintain tonicity, and cells swell and expand. Membrane proteins > are fixed and crosslinked to neighboring cells in the swollen position. > Later, the cells stabilize and contract, and pull other cells with them. > The result is gross shrinkage, local distortion and torn membranes with > lost cellular contents from some cells. > > Tissue Shrinkage can be completely prevented if the extracellular fluid > with ions is replaced by a nonionic isotonic solution that cannot > enter the > cells. This can be accomplished by prewash with 5% (isotonic) sucrose in > distilled water. Start the prewash at low pressure, and pump up to 300 > mmHg > to break the blood brain barrier and wash the extracellular fluid (and > red > blood cells). Switch shortly thereafter to fixative. The perfusion takes > the same time as the traditional procedure, and accomplishes the > favorable > results bulleted above. > > So if I'd like to try out the perfusion techniques that are made > simple by > the Perfusion one system, what would you suggest: first do a > physiological saline solution, followed by a prewash with 5% sucrose and > then PFA, or go directly to 5% sucrose then PFA? > > thanks for your advice, > > Nancy > > -- -- ********************************************** 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 ********************************************** -- -- ********************************************** 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 ohenry <@t> dfw.net Thu Oct 9 12:29:39 2003 From: ohenry <@t> dfw.net (Susan Owens) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] GI bx's and Warthin Starry Message-ID: <000901c38e8b$1b5a31c0$c9f763d8@Nationwide.net> Over the years we also had this problem(from time to time), and we will have it again I'm sure whenever they get new people for the GI rooms. But hopefully not.... The problem of not staining around the edges on silver stains comes from the nurse allowing the GI bx's to lay out too long before putting them into the Hollands fixative instead of putting them immediately into the fixative. The regular H&E and the AB/PAS were not effected, only our silver stain,Steiner/Steiner... What allot of the nurses/assistants couldn't seem to understand was the real need to put the bx IMMEDIATELY into the fixative... They wanted to take the bx and lay them out for a count of the number they have since they were hard to count after being put into the bottle. So I'm told....Of course that answer didn't account for the single piece that was allowed to stay out....The word "immediately" should be clear, but to some a slight delay is still 'immediate'....And of course a 'slight delay' to one person may be a coffee break to another. hehe. Hope this helps. Susan Owens,HT ---------------------------------------------------------------------------- ---- Message: 1 From: "celia" To: Date: Wed, 8 Oct 2003 17:23:06 -0500 Subject: [Histonet] warthin starry Hello Everyone, I have put this question out once before and I didn't get much for a = response so I thought I would try putting it out there one more time. I = have an ongoing problem with my Warthin-Starry stain. The peripheral = edges of some of the gastric biopsies are not staining. I am also = experiencing this same non-staining with the Steiner stain on the = Ventana stainer. I have tried reducing the temp on the processor down = from 60c to 58c thinking they might be "cooking" on the processor (the = H&E's look fine) as well as checking the ph of my solutions. This did = not work. Could this be a handling problem prior to fixation in 10% = NBF? I am at my wits end. Any suggestions will be greatly appreciated. Thanks in advance. Celia, HT SVH, Sioux Falls, SD From Jacquie.Mack <@t> CLS.ab.ca Thu Oct 9 12:32:36 2003 From: Jacquie.Mack <@t> CLS.ab.ca (Mack, Jacquie) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] plastination thank you! Message-ID: <30C050525B881C4AAFF41E6D16543E6802541D22@mail3.cls.ab.ca> Thank you to everyone with helpful websites and references for my plastination questions! Once again, the histonet is a wonderful tool and I"m thankful for such helpful technologists "nearby"! Sincerely, Jacqueline Mack Tech III Anatomic Pathology Foothills Medical Center Calgary Laboratory Services ph (403) 944-4162 fax (403) 270-4093 e mail: Jacquie.Mack@CLS.ab.ca -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031009/eb53567b/attachment.htm From nick.kirk3 <@t> btopenworld.com Thu Oct 9 12:46:43 2003 From: nick.kirk3 <@t> btopenworld.com (Nick Kirk) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Need silver stain for granules in adrenals In-Reply-To: <20031009150458.75311.qmail@web80109.mail.yahoo.com> Message-ID: What you are trying to demonstrate are Chromaffin cells of the adrenal glands so a Masson-Fontana should work, if not try a Schmorl or carry out the Chromaffin reaction If you use Iodate oxidation you can distinguish between Adrenaline and noradrenalin fairly easily as iodates oxidise noradrenaline much quicker than they do adrenaline. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Nancy Maronto Sent: 09 October 2003 16:05 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Need silver stain for granules in adrenals Hi, Could anyone suggest a stain for noradrenalin and adrenalin granules in paraffin embedded adrenals. We were asked to do a gremilius stain and it did not stain the granules. Are these slides lost or can we re stain them? We usually do a recut of the block, but it is asked if we could use the same tissue slide. The stain actually worked, but not for the granules. From what we read a Fontana-masson should work. There probably is a better stain our there for this specific target. Can anyone with some experience with staining these granules share their staining protocol. At this time the request is not for immuno staining. Thanks for your help. Nancy Maronto -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031009/763d4d9a/attachment.htm From rfail <@t> toolkitmail.com Thu Oct 9 12:20:40 2003 From: rfail <@t> toolkitmail.com (rfail@toolkitmail.com) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] GFAP Message-ID: <01L1MK5BIVKM8ZG0RB@SMTP00.InfoAve.Net> I would like to know the preferred clone for this Antibody Rena Fail From ohenry <@t> dfw.net Thu Oct 9 13:03:22 2003 From: ohenry <@t> dfw.net (Susan Owens) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Haze over GI bx's on H&E's Message-ID: <002f01c38e8f$b103f540$c9f763d8@Nationwide.net> We also experienced poor H&E staining on GI's fixed in Hollands...We solved it by doing the following. 1. BEFORE the Hollands fixed bx's are loaded onto the processor they are given a 10+ minutes wash in running tap water....This is important. 2. They are stained separately from the other(routine) tissues, but on the same staining line. We give another 10 wash before the Hemo(This also is important)......We also increase the time in Hemo for the bx's, 2 times what the over tissues get. We stain progessively. Try it, you have nothing to loose but a little time. You will have to play some with your staining times, but I believe it will solve the problem, it did for us. The last time we had a 'haze' problem the gross room people admitted they were rushed and rinsed, not washed the bx's before loading on the processors. Susan Owens,HT ohenry@dfw.net fax: 817-548-9876 "A bad day at the dog show is better then a good day at work!" From cherylsgarden <@t> mindspring.com Thu Oct 9 13:07:29 2003 From: cherylsgarden <@t> mindspring.com (cherylsgarden@mindspring.com) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] fat-vacuole type artifact on bronch & lung bx Message-ID: <15394227.1065722849850.JavaMail.root@wamui10.slb.atl.earthlink.net> Help! All of a sudden we are experiencing an artifact on tissues such as lung biopsies and bronch biopsies that appear to be fat globules on most of the Hollandes fixed specimens. We had upped a few times on our automatic processor and use vaccuum and pressure and a little heat with ProSoft and ProClear as the solvents. It is programmed with a biopsy program for small tissues--roughly 8 hours. This type of tissue is the only one affected from that processor. We do process in sponges and it's a little like the trabeculations you can get as a sponge artifact except that it's through the whole tissue and restricted to lung-type tissue. Anything you guys know that might be causing this--I'd appreciate your suggestions. Cheryl Kerry Niday cherylniday@centura.org Penrose Hospital Colorado Springs. From Bauer.Karen <@t> mayo.edu Thu Oct 9 13:37:34 2003 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Ventana BenchMark IP stainer Message-ID: <8C6E05FA69571948B461F1327CBB893E16C2A3@LMMAIL2> Hi, One of our Pathologists is unhappy with some of the results from our BenchMark IP stainer. Not all, but some tissues fall off and there are stains where the tissue seems to be "burnt" or washed out. We realize that fixation has a lot to do with the way a stain can turn out, and I've told him this many times. I'm sure that everyone has had to deal with "Hurry and get this on the Processor tonight" and then the next morning we are supposed to make everything cut, stain, and look perfect. This isn't always the case. There will be well fixed tissue that will look and stain the same way. We were having this problem with our Ventana Nexes stainer, but after some micron and heating time adjustments, things started looking good again. Now that we've upgraded to the BenchMark, it's like we are starting from scratch again. We've had it since July of this year and our Pathologist would really like our Nexes back. The thought of changing all of our IP procedures again does not sound appealing to me. Right now, this is the way we cut and prepare our slides for IP staining on the BenchMark: Cut sections at 4 to 5 microns and place on a positive and negative control slide. (We use the charged slides with the red control box on them.) We let them air dry for 30 minutes and then we place them in a 60 degree C oven for 30 minutes to 1 hour. We then place them on the BenchMark for staining. The stains that we are having the most troubles with are the ones using the CC1. I've checked our buffers and the pH is fine. We're having difficulty with our Pathway Her 2 also. This uses the CC2 and it ends up looking washed out. If we keep it at the mild CC2, there's staining but it's light. Standard CC2 looks like the tissue is "eaten up or burnt out", and Extended CC2 makes the tissue pretty much fall off. This only happens to the pt. tissue. Our control tissue seems to stain fine. (Pt. tissue and control tissue are on the same slide.) I've talked to our Ventana Rep and their technical personnel and no one has given me any ideas that we haven't tried already. Can anybody give me some hints or solutions to these problems? Has anyone else out there had these problems? Thanks!! Karen L. Bauer Histology Department Luther Hospital 715-838-3205 ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. From mcauliff <@t> umdnj.edu Thu Oct 9 13:51:15 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] GFAP In-Reply-To: <01L1MK5BIVKM8ZG0RB@SMTP00.InfoAve.Net> References: <01L1MK5BIVKM8ZG0RB@SMTP00.InfoAve.Net> Message-ID: <3F85AE23.4070403@umdnj.edu> I have always had excellent results with DAKO's rabbit anti-cow GFAP on both FFPE sections and frozen sections. I use a Vector ABC Elite kit with DAB. Geoff rfail@toolkitmail.com wrote: >I would like to know the preferred clone for this Antibody >Rena >Fail > > >_______________________________________________ >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 mprice26 <@t> juno.com Thu Oct 9 13:29:49 2003 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Re: Decontaminating cryostat after TB infected fresh tissue Message-ID: <20031009.113041.4667.17512@webmail21.nyc.untd.com> Hi, histonetters, I am in need of an procedure for decontaminating the cryostat after doing an frozen on infectious fresh tissue. If anyone can share one with me. 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 sa.drew <@t> hosp.wisc.edu Thu Oct 9 14:41:20 2003 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Ventana BenchMark IP stainer Message-ID: We use both the Nexes and Benchmark and we've not really experienced your particular problem on a large scale- but I might wonder if there is a problem w/ reagent slide volume during the depar/CC step. I'm sure you've probably visually checked the priming of those reagents, but maybe a VMS rep could check the volume delivered, or tell you how to. Do you have the same problem with immunos that don't require any CC? We use the same control slide set up and all, but we don't routinely put our slides designated for the BMK in the oven before putting them on-I'm not sure it's necessary (but I also don't think that's the problem. Sally Ann Drew-MT(ASCP) Univ. of Wisconsin Hospital & Clinics IHC/ISH Laboratory 600 Highland Ave. VAH-DM223 Madison, WI 53792-2472 608-265-6596 sa.drew@hosp.wisc.edu -----Original Message----- From: Bauer, Karen [mailto:Bauer.Karen@mayo.edu] Sent: Thursday, October 09, 2003 1:38 PM To: Histonet (E-mail) Subject: [Histonet] Ventana BenchMark IP stainer Hi, One of our Pathologists is unhappy with some of the results from our BenchMark IP stainer. Not all, but some tissues fall off and there are stains where the tissue seems to be "burnt" or washed out. We realize that fixation has a lot to do with the way a stain can turn out, and I've told him this many times. I'm sure that everyone has had to deal with "Hurry and get this on the Processor tonight" and then the next morning we are supposed to make everything cut, stain, and look perfect. This isn't always the case. There will be well fixed tissue that will look and stain the same way. We were having this problem with our Ventana Nexes stainer, but after some micron and heating time adjustments, things started looking good again. Now that we've upgraded to the BenchMark, it's like we are starting from scratch again. We've had it since July of this year and our Pathologist would really like our Nexes back. The thought of changing all of our IP procedures again does not sound appealing to me. Right now, this is the way we cut and prepare our slides for IP staining on the BenchMark: Cut sections at 4 to 5 microns and place on a positive and negative control slide. (We use the charged slides with the red control box on them.) We let them air dry for 30 minutes and then we place them in a 60 degree C oven for 30 minutes to 1 hour. We then place them on the BenchMark for staining. The stains that we are having the most troubles with are the ones using the CC1. I've checked our buffers and the pH is fine. We're having difficulty with our Pathway Her 2 also. This uses the CC2 and it ends up looking washed out. If we keep it at the mild CC2, there's staining but it's light. Standard CC2 looks like the tissue is "eaten up or burnt out", and Extended CC2 makes the tissue pretty much fall off. This only happens to the pt. tissue. Our control tissue seems to stain fine. (Pt. tissue and control tissue are on the same slide.) I've talked to our Ventana Rep and their technical personnel and no one has given me any ideas that we haven't tried already. Can anybody give me some hints or solutions to these problems? Has anyone else out there had these problems? Thanks!! Karen L. Bauer Histology Department Luther Hospital 715-838-3205 ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From la.sebree <@t> hosp.wisc.edu Thu Oct 9 14:45:12 2003 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Ventana BenchMark IP stainer Message-ID: Karen, We find that once you go to mild CC1 or CC2, you may have problems with tissue morphology and/or tissue loss yet plain CC1 or CC2 may not be long enough retrieval time. Sometimes if we need more HIER but don't want to extend our time to the mild setting we'll add a weak protease (P2 or P3) for a short time to enhance the retrieval effect without further tissue damage. Also, the reason you're probably getting worse results with CC1 is that it is the harsher of the two buffers (EDTA) at a higher pH than CC2. We generally do not oven dry slides going on the Benchmark; just make sure they are well air-dried. So maybe your slides are just being exposed to too much heat between the oven and deparaffinization on the instrument. So try backing off on a few things that may be damaging your tissues and see if that helps. Feel free to contact us with specific questions as we have dealt with a number of the same issues. Linda A. Sebree, HT(ASCP) University of Wisconsin Hospital & Clinics IHC/ISH Laboratory A4/204-2472 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: Bauer, Karen [mailto:Bauer.Karen@mayo.edu] Sent: Thursday, October 09, 2003 1:38 PM To: Histonet (E-mail) Subject: [Histonet] Ventana BenchMark IP stainer Hi, One of our Pathologists is unhappy with some of the results from our BenchMark IP stainer. Not all, but some tissues fall off and there are stains where the tissue seems to be "burnt" or washed out. We realize that fixation has a lot to do with the way a stain can turn out, and I've told him this many times. I'm sure that everyone has had to deal with "Hurry and get this on the Processor tonight" and then the next morning we are supposed to make everything cut, stain, and look perfect. This isn't always the case. There will be well fixed tissue that will look and stain the same way. We were having this problem with our Ventana Nexes stainer, but after some micron and heating time adjustments, things started looking good again. Now that we've upgraded to the BenchMark, it's like we are starting from scratch again. We've had it since July of this year and our Pathologist would really like our Nexes back. The thought of changing all of our IP procedures again does not sound appealing to me. Right now, this is the way we cut and prepare our slides for IP staining on the BenchMark: Cut sections at 4 to 5 microns and place on a positive and negative control slide. (We use the charged slides with the red control box on them.) We let them air dry for 30 minutes and then we place them in a 60 degree C oven for 30 minutes to 1 hour. We then place them on the BenchMark for staining. The stains that we are having the most troubles with are the ones using the CC1. I've checked our buffers and the pH is fine. We're having difficulty with our Pathway Her 2 also. This uses the CC2 and it ends up looking washed out. If we keep it at the mild CC2, there's staining but it's light. Standard CC2 looks like the tissue is "eaten up or burnt out", and Extended CC2 makes the tissue pretty much fall off. This only happens to the pt. tissue. Our control tissue seems to stain fine. (Pt. tissue and control tissue are on the same slide.) I've talked to our Ventana Rep and their technical personnel and no one has given me any ideas that we haven't tried already. Can anybody give me some hints or solutions to these problems? Has anyone else out there had these problems? Thanks!! Karen L. Bauer Histology Department Luther Hospital 715-838-3205 ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Thu Oct 9 16:14:48 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Need silver stain for granules in adrenals In-Reply-To: References: Message-ID: <3F85CFC8.9020308@umdnj.edu> I don't think you can get a good chromaffin reaction on material already fixed with formalin. Fontana-Masson should be fine. John Kiernan published a chromate-dichromate fix for chromaffin tissues, it may be in his book, along with other recommendations. Geoff Nick Kirk wrote: > What you are trying to demonstrate are Chromaffin cells of the adrenal > glands so a Masson-Fontana should work, if not try a Schmorl or carry > out the Chromaffin reaction > If you use Iodate oxidation you can distinguish between Adrenaline and > noradrenalin fairly easily as iodates oxidise noradrenaline much > quicker than they do adrenaline. > > Nick Kirk > Histopathology > Hinchingbrooke Hospital > Huntingdon > England > > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Nancy > Maronto > Sent: 09 October 2003 16:05 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Need silver stain for granules in adrenals > > Hi, > Could anyone suggest a stain for noradrenalin and adrenalin > granules in paraffin embedded adrenals. We were asked to do a > gremilius stain and it did not stain the granules. Are these > slides lost or can we re stain them? We usually do a recut of the > block, but it is asked if we could use the same tissue slide. The > stain actually worked, but not for the granules. > > From what we read a Fontana-masson should work. There probably is > a better stain our there for this specific target. Can anyone > with some experience with staining these granules share their > staining protocol. > > At this time the request is not for immuno staining. > > Thanks for your help. > > Nancy Maronto > -- -- ********************************************** 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 ********************************************** -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031009/52ae98d8/attachment.htm From peptolab <@t> hamptons.com Thu Oct 9 16:36:43 2003 From: peptolab <@t> hamptons.com (peptolab) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Movat Green connective tissue Message-ID: <007801c38ead$70164560$e176bd18@JEFF> I invented a Movat modification in 1972 and so have a little experience with this stain. Green connective tissue can stem from two causes. Sometimes the connective tissue is green because it is particularly rich in alcianophilic ground substance (extracellular matrix mucopolysaccharides) and is lightly collagenized, producing only fibrils rather than thick collagen bundles. You remeber from art class that (alcian) blue plus (saffron) yellow equals green and there you go. If your saffron is tired or even slightly waterlogged you might see this too but densley ciollagenized tissue would be a very pale yellow not green. I used to keep a layer of Drierite at the bottom of the stock bottle and dehydrate with three changes of absolute before and after the saffron. Hope this helps. Jeff Silverman HT HTL QIHC Pathologists' Assistant Southside Hopsital Bay Shore NY USA From peptolab <@t> hamptons.com Thu Oct 9 16:46:07 2003 From: peptolab <@t> hamptons.com (peptolab) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Terpenes vs xilol Message-ID: <008001c38eae$c1a9cbd0$e176bd18@JEFF> Lina- Funny I use it just the opposite. I find xykene (xilol) better for tissue processing and limonene better for staining. I use the non toxic limonene in all the dewax and clearing baths on the H and E line and use only one xilol at the end to cover slip from since it evasporates faster. I find xilol is better at clearing the blocks and limonene leaves some blocks oily. I use the limonene to purge clean the tissue processor too- it works fine on my Leica TP 1050 and on my old VIP. This eliminates a lot of xylene exposure in my lab. For more setails see- Silverman JS. 1999. d-Limonene: A serviceable and safe routine clearing agent. Microscopy Today #99-10, p. 18. Jeff Silverman H HTL QIHC Pathologists' Assistant Southside Hopistal Bay Shore NY USA From Robert.Fauck <@t> ccdhb.org.nz Thu Oct 9 17:19:50 2003 From: Robert.Fauck <@t> ccdhb.org.nz (Robert Fauck) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] CD15 and CD3 Message-ID: Hi Could any one suggest how to enhance the "crispiness" of CD3 and CD15"staining"around the lymphocytes? We have some problem with both DAKO and NOVOCASTRA for these two markers. Thanks, Robert Fauck C&C DHB Secure Mail Server ******************************************************************************** This email or attachment(s) may contain confidential or legally privileged information intended for the sole use of the addressee(s). Any use, redistribution, disclosure, or reproduction of this message, except as intended, is prohibited. If you received this email in error, please notify the sender and remove all copies of the message, including any attachments. Any views or opinions expressed in this email (unless otherwise stated) may not represent those of Capital and Coast District Health Board. (AC_S001) [INFO] -- Virus Manager: No Viruses were detected in this message. ******************************************************************************** From RFail <@t> Charleston.net Thu Oct 9 17:31:46 2003 From: RFail <@t> Charleston.net (Rena Fail) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] CD15 and CD3 In-Reply-To: Message-ID: <00db01c38eb5$2015a4e0$d511a6a5@rena> Robert, We use DAKO's CD3 and CD15. CD3 almost every day. We get nice crisp staining with both. Rena Fail -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Robert Fauck Sent: Thursday, October 09, 2003 6:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CD15 and CD3 Hi Could any one suggest how to enhance the "crispiness" of CD3 and CD15"staining"around the lymphocytes? We have some problem with both DAKO and NOVOCASTRA for these two markers. Thanks, Robert Fauck C&C DHB Secure Mail Server ************************************************************************ ******** This email or attachment(s) may contain confidential or legally privileged information intended for the sole use of the addressee(s). Any use, redistribution, disclosure, or reproduction of this message, except as intended, is prohibited. If you received this email in error, please notify the sender and remove all copies of the message, including any attachments. Any views or opinions expressed in this email (unless otherwise stated) may not represent those of Capital and Coast District Health Board. (AC_S001) [INFO] -- Virus Manager: No Viruses were detected in this message. ************************************************************************ ******** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RFail <@t> Charleston.net Thu Oct 9 17:54:06 2003 From: RFail <@t> Charleston.net (Rena Fail) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] CD15 and CD3 more info In-Reply-To: Message-ID: <00dc01c38eb8$3f0f4d80$d511a6a5@rena> Sorry I should have added both are at a dilution of 1:100, both are heat retrieved with a buffer pH6.0 ,in a steamer for 20 minutes with a 10 minute cool down Rena -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Robert Fauck Sent: Thursday, October 09, 2003 6:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CD15 and CD3 Hi Could any one suggest how to enhance the "crispiness" of CD3 and CD15"staining"around the lymphocytes? We have some problem with both DAKO and NOVOCASTRA for these two markers. Thanks, Robert Fauck C&C DHB Secure Mail Server ************************************************************************ ******** This email or attachment(s) may contain confidential or legally privileged information intended for the sole use of the addressee(s). Any use, redistribution, disclosure, or reproduction of this message, except as intended, is prohibited. If you received this email in error, please notify the sender and remove all copies of the message, including any attachments. Any views or opinions expressed in this email (unless otherwise stated) may not represent those of Capital and Coast District Health Board. (AC_S001) [INFO] -- Virus Manager: No Viruses were detected in this message. ************************************************************************ ******** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nick.kirk3 <@t> btopenworld.com Thu Oct 9 20:43:55 2003 From: nick.kirk3 <@t> btopenworld.com (Nick Kirk) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Need silver stain for granules in adrenals In-Reply-To: <3F85CFC8.9020308@umdnj.edu> Message-ID: You could try secondary fixing the tissue in chromate solution to get the Chromaffin reaction, it usually works. Nick Kirk -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Geoff McAuliffe Sent: 09 October 2003 22:15 To: Nick Kirk Cc: Histonet Subject: Re: [Histonet] Need silver stain for granules in adrenals I don't think you can get a good chromaffin reaction on material already fixed with formalin. Fontana-Masson should be fine. John Kiernan published a chromate-dichromate fix for chromaffin tissues, it may be in his book, along with other recommendations. Geoff Nick Kirk wrote: What you are trying to demonstrate are Chromaffin cells of the adrenal glands so a Masson-Fontana should work, if not try a Schmorl or carry out the Chromaffin reaction If you use Iodate oxidation you can distinguish between Adrenaline and noradrenalin fairly easily as iodates oxidise noradrenaline much quicker than they do adrenaline. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Nancy Maronto Sent: 09 October 2003 16:05 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Need silver stain for granules in adrenals Hi, Could anyone suggest a stain for noradrenalin and adrenalin granules in paraffin embedded adrenals. We were asked to do a gremilius stain and it did not stain the granules. Are these slides lost or can we re stain them? We usually do a recut of the block, but it is asked if we could use the same tissue slide. The stain actually worked, but not for the granules. From what we read a Fontana-masson should work. There probably is a better stain our there for this specific target. Can anyone with some experience with staining these granules share their staining protocol. At this time the request is not for immuno staining. Thanks for your help. Nancy Maronto -- -- ********************************************** 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 ********************************************** -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031010/e5a5dd19/attachment.htm From GREYTRUNK <@t> aol.com Thu Oct 9 20:48:40 2003 From: GREYTRUNK <@t> aol.com (GREYTRUNK@aol.com) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Purple Haze..... Message-ID: <67.1a320c89.2cb769f8@aol.com> We had this same problem. We had control over everything, too. We stopped using Sta-On, etrc. ANd we still had the problem - haziness, it looked like the mucin was staining, but we also had the fuzziness on non-GI's as well. It turned out to be two things--the hematoxylin we were using and the timing of it and the temperature of the water on the stainer--it was too cold. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031009/2544bb8a/attachment.htm From kwuny <@t> email.cs.nsw.gov.au Thu Oct 9 20:57:24 2003 From: kwuny <@t> email.cs.nsw.gov.au (Young Kwun) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] DiaSorin company? Message-ID: <001201c38ed1$d9ba4c40$a6e04c98@lab0220.cs.nsw.gov.au> Dear Histonetters, I would like to know about a company called DiaSorin Inc. Their website is not on and I want to know their email address. They used to make excellent immunofluorescence antibodies, but we cannot get them at the moment. Thank you in advance. Young Kwun Senior Hospital Scientist Dept. of Anatomical Pathology Concord Hospital Concord NSW 2139 Australia Tel)61-2-9767-6075 Fax)61-2-9767-8427 kwuny@email.cs.nsw.gov.au "the universe we observe has precisely the properties we should expect if there is, at bottom, no design, no purpose, no evil and no good, nothing but blind, pitiless indifference." "To err is human, to forgive is even more human" The information contained in this message is intended for the named addressee only, and is confidential to the sender and intended recipient. If you are not the named addressee please do not copy, distribute, take any action reliant on, or disclose anything in this E-mail message to any other person or organisation. If you have received this message in error please delete the email and notify me immediately. Views expressed in this message are those of the individual sender and are not necessarily the views of Central Sydney Area Health Service. From thallada <@t> noch.org Fri Oct 10 09:19:02 2003 From: thallada <@t> noch.org (Hallada, Teri) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] RE: Immunos Message-ID: Hi all, I have an inquiry about immunos. We have differing opinions here at our institution. We have one pathologist that likes to utilize immunos in diagnosing cases and one that insists that they are not required to make a diagnosis. We currently send all of our immunos out to be stained and read them here upon return. We are considering trying to bring them in house, but are concerned with the volume that we are ordering. We are a small community hospital, but have a pretty good outpatient clientele also. I was wondering if I could get some feedback from the group about the importance of utilizing immunos and volumes that are being ordered. Also, do any of you use certain immunos as part of your protocol for specific cases (other than breast). Thank you, Teri Hallada BS MT CT (ASCP) thallada@noch.org > ___________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From weimer_r_v <@t> hotmail.com Fri Oct 10 09:39:19 2003 From: weimer_r_v <@t> hotmail.com (r v weimer) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] DiaSorin company? Message-ID: At the web site www.antibodies-probes.com under 'Company Search' there are listed 5600+ biotech companies including 600+ antibody manufacturers and suppliers. Bob >From: "Young Kwun" >Reply-To: >To: "Histonet" >Subject: [Histonet] DiaSorin company? >Date: Fri, 10 Oct 2003 11:57:24 +1000 > >Dear Histonetters, >I would like to know about a company called DiaSorin Inc. Their website is >not on and I want to know their email address. They used to make excellent >immunofluorescence antibodies, but we cannot get them at the moment. >Thank you in advance. > > > > > > >Young Kwun >Senior Hospital Scientist >Dept. of Anatomical Pathology >Concord Hospital >Concord NSW 2139 Australia >Tel)61-2-9767-6075 >Fax)61-2-9767-8427 >kwuny@email.cs.nsw.gov.au > >"the universe we observe has precisely the properties we should expect if >there is, at bottom, no design, no purpose, no evil and no good, nothing >but >blind, pitiless indifference." >"To err is human, to forgive is even more human" > >The information contained in this message is intended for the named >addressee only, and is confidential to the sender and intended recipient. >If >you are not the named addressee please do not copy, distribute, take any >action reliant on, or disclose anything in this E-mail message to any other >person or organisation. If you have received this message in error please >delete the email and notify me immediately. Views expressed in this message >are those of the individual sender and are not necessarily the views of >Central Sydney Area Health Service. > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Get 10MB of e-mail storage! Sign up for Hotmail Extra Storage. http://join.msn.com/?PAGE=features/es From SBarnes <@t> elch.org Fri Oct 10 10:07:57 2003 From: SBarnes <@t> elch.org (Sue Barnes) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] RE: Immunos Message-ID: <3F74CB2B769A5843A0C746D3F13256FD1279F2@ELCH2> Hi, We are also a small community hospital, we have many different antibodies available at all times. We routinely do: Liver biopsy: HBSAG, HCV, Vimentin, Kermix, S100, Iron, PAS, PASD, Retic, Trichrome Bladder tumor: Actin (MS) Gastric Biopsies: H Pylori, Alcian Bl, Giemsa Esophageal Biopsies: H Pylori, PAS/Alcian Blue, Giemsa All lymph nodes from cancer (colon, breast) Pan Keratin Our pathologist is in his 30's, older pathologist ordered lots also but not as specific Hope this helps -----Original Message----- From: Hallada, Teri [mailto:thallada@noch.org] Sent: Friday, October 10, 2003 10:19 AM To: Histonet-LIST Subject: [Histonet] RE: Immunos Hi all, I have an inquiry about immunos. We have differing opinions here at our institution. We have one pathologist that likes to utilize immunos in diagnosing cases and one that insists that they are not required to make a diagnosis. We currently send all of our immunos out to be stained and read them here upon return. We are considering trying to bring them in house, but are concerned with the volume that we are ordering. We are a small community hospital, but have a pretty good outpatient clientele also. I was wondering if I could get some feedback from the group about the importance of utilizing immunos and volumes that are being ordered. Also, do any of you use certain immunos as part of your protocol for specific cases (other than breast). Thank you, Teri Hallada BS MT CT (ASCP) thallada@noch.org > ___________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From brett_connolly <@t> merck.com Fri Oct 10 10:51:41 2003 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] CCR-1 antibody Message-ID: Looking for suppliers, any tips/requirements for use in IHC on frozens...human. Thanks, 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 From Kay.Cullen <@t> umassmed.edu Fri Oct 10 11:17:35 2003 From: Kay.Cullen <@t> umassmed.edu (Cullen, Kay) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] acrosome Message-ID: I am looking for a method to stain the acrosome of mouse sperm. My boss thinks he remembers using bromocresol green but I can find no references. From Tiffany.L.Sheffield <@t> uth.tmc.edu Fri Oct 10 11:40:05 2003 From: Tiffany.L.Sheffield <@t> uth.tmc.edu (Tiffany L Sheffield) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] IL-1 and IL-6 question Message-ID: <3F86E0E5.2067D7C8@uth.tmc.edu> Hello Fellow Histonetters! I am venturing into a new area of Ab's and I wondered if you all could offer some advice. I am wanting to run IHC testing using Interleukin 1&6 on mouse distal femurs. Is there specific IL-1&6 Ab's for the use in bone or can I use the same ones that I have used in soft tissues. I was not sure...... Thank you, Tiff -------------- next part -------------- A non-text attachment was scrubbed... Name: Tiffany.L.Sheffield.vcf Type: text/x-vcard Size: 374 bytes Desc: Card for Tiffany Sheffield-Lopez Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031010/377fb00a/Tiffany.L.Sheffield.vcf From MGomez <@t> ameripath.com Fri Oct 10 11:44:59 2003 From: MGomez <@t> ameripath.com (MGomez@ameripath.com) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Histology Textbooks Message-ID: Dear Histonetters: I'm looking for Histology Textbooks I can use as a reference to assist me with Lab. Procedures and Protocols. Any recommendations? Thank you very much, Milton 801-256-0040 x205 From ccdub <@t> earthlink.net Fri Oct 10 11:42:59 2003 From: ccdub <@t> earthlink.net (Cindy/Rick DuBois) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Cyto Preps Message-ID: <28865491.1065804180429.JavaMail.root@gonzo.psp.pas.earthlink.net> We are interested in what reagents/procedures other labs are using to process bloody body fluids. What are you using to lyse the RBC's? We currently use plasma/thrombin, but our cytotechs are having a difficult time reading the slides. Does anyone know of a rapid method that does not interfere with immuno stains? Cindy DuBois, HT DELTA PATHOLOGY ASSOC. STOCKTON CA. From ljb <@t> medicine.wisc.edu Fri Oct 10 11:04:34 2003 From: ljb <@t> medicine.wisc.edu (LaCinda Burchell) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] we need an antibody which is specific for smooth muscle Message-ID: Dear Histonetters and Vendors, I have two questions. 1. Does myosin heavy chain stain smooth muscle cells? 2. Can anyone tell me what is exclusive for smooth muscle, not myofibroblasts or fibroblasts? Thanks Much, Cindy B LaCinda Burchell, BA, AS, HT(ASCP) University of Wisconsin-Madison, Medical School Asthma and Allergy Research IHC Lab 600 Highland Ave. CSC K4/913 Madison, Wisconsin 53792 Phone: 608-262-3518 FAX: 608-263-3746 From TJasper <@t> smdc.org Fri Oct 10 12:32:32 2003 From: TJasper <@t> smdc.org (Jasper, Thomas) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Immunos Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA23FCD4@harrier.ntcampus.smdc.org> Dear Teri, While it is true that a pathologist can diagnose without immunos (after all diagnostic pathology existed pre-immunohistochemistry) it is uncommon for a pathologist not to use immunos, even in a limited capacity. Cases of a certain nature may not call for immunos, but aside from that most pathologists choose to employ them. The fact that your immunos go out to be done (technical) and return to be interpreted (professional) tells me that despite having a pathologist that does not require them, there is enough work (and diagnostic understanding by the other path) that it may be feasible to do your own. Gauging volumes has merit, but you really need to figure out what works best for you. What I mean by that is calculating a true cost. How much do you pay in technical fees to have the immunos sent out? Does this involve other departments and is there a cost associated with diagnostic delay? As long as you do the interpretation your path should be capturing the professional charges. Compare the cost of technical send-outs to the cost of running in-house. This is where your volumes would come in. Volumes and frequency of antibody type. Most suppliers of immunohistochemical products will help you calculate this. They have an interest in being as accurate as possible because it does them no good to have you as a customer if there truly is no need on your part. Obviously some ABs cost more than others and then there are ancillary costs for reagents etc. Many companies today (Dako, Ventana etc.) will get you up and running. You can generally get an equipment lease package with a reagent rental agreement. Or you could opt for manual kits, although I prefer automation as it really eliminates variables. I also believe that automation helps with troubleshooting and simplifies QC for your paths and regulating agencies. I'm sure there are many monitoring this list server that would be happy to provide you with more details on such options. Now for my own non-pathologist opinion on the importance of immunos...as an AP Coordinator and someone who has been working in histopathology for almost 20 years I believe immunohistochemistry to be extremely important. Antigen/antibody complexes are just too good a bio tool to NOT use. The day is surely coming when the last pathologist to not use immunos will retire. Onto your question about protocols other than breast...we run S-100 and Melan A with every sentinel node case for melanoma (protocol). We don't have any other protocol that is automatically ordered at the grossing bench (other than sentinel node breast cases), however our paths like certain lymphoma panels (CD markers) which are ordered on a case by case basis. Generally a lot of ABs will be run together, CK7 and 20, Kappa/Lambda, etc. but here again you really need to look at your unique service, what your pathologist wants and if/how you can provide it. Good luck to you! Thomas G. Jasper Anatomic Pathology Coordinator SMDC Duluth, MN 55805 (218) 786-4510 tjasper@smdc.org From Cheryl.Clarke <@t> fraserhealth.ca Fri Oct 10 12:46:26 2003 From: Cheryl.Clarke <@t> fraserhealth.ca (Clarke, Cheryl) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] unsubscribe Message-ID: please unsubscibe From Rcartun <@t> harthosp.org Fri Oct 10 11:25:25 2003 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] RE: Immunos Message-ID: Obviously, immunohistochemical stains are not required for the vast majority of specimens that we see in the surgical pathology laboratory, but, if I had a completed lesion removed, I would not want it sent to your pathologist who claims that immunohistochemical stains are not required for diagnosis! Richard Cartun >>> "Hallada, Teri" 10/10/03 10:19AM >>> Hi all, I have an inquiry about immunos. We have differing opinions here at our institution. We have one pathologist that likes to utilize immunos in diagnosing cases and one that insists that they are not required to make a diagnosis. We currently send all of our immunos out to be stained and read them here upon return. We are considering trying to bring them in house, but are concerned with the volume that we are ordering. We are a small community hospital, but have a pretty good outpatient clientele also. I was wondering if I could get some feedback from the group about the importance of utilizing immunos and volumes that are being ordered. Also, do any of you use certain immunos as part of your protocol for specific cases (other than breast). Thank you, Teri Hallada BS MT CT (ASCP) thallada@noch.org > ___________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From drbugge <@t> msn.com Fri Oct 10 12:58:58 2003 From: drbugge <@t> msn.com (Dawn Bugge) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] online histology training Message-ID: I was just wondering if anyone has heard of an online histology certification training from Harford. I have been trying to find it online, but I haven't been successful. Thank you very much Dawn _________________________________________________________________ Get 10MB of e-mail storage! Sign up for Hotmail Extra Storage. http://join.msn.com/?PAGE=features/es From Wijaya.Martanto <@t> chbe.gatech.edu Fri Oct 10 13:34:19 2003 From: Wijaya.Martanto <@t> chbe.gatech.edu (Martanto, Wijaya) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] (no subject) Message-ID: <5D3DB84BB6F5E548AAC9FFEDC310FD091BB8F3@chicken-boo.che.gatech.edu> hi, I posted this question earlier but I did not get many responses, so I would like to ask a similar but more general question: I was looking for a dye which is commonly used to mark aqueous part of tissue but does not have strong binding affinity within tissue. Specifically, I was looking for a dye which does not bind locally within animal skin in vitro (tissue marking dye will bind locally within skin). I think the dye should be hydrophilic to allow 'spreading' within the tissue. I am planning to inject the dye into the animal skin in vitro and would like to see the dye distribution within the skin. Currently, I am using tissue marking dye (which I think it's hydrophobic) and it binds locally within the skin (no diffusion/spreading). I was wondering if you have some suggestion on which commercially-available dye I should use. Any replies can be directed to my email address shown below: Thanks. Sincerely, Wijaya Martanto wijaya.martanto@chbe.gatech.edu -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031010/f3c01233/attachment.htm From GDawson <@t> Milw.Dynacare.com Fri Oct 10 13:44:04 2003 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Non mouse CD20 Message-ID: All, Does anyone out there use a good CD20 IHC for FFPE blocks that was NOT made in a mouse? I would really like a goat or rabbit CD20 but have not been able to find one. Thanx in Advance, Glen Dawson From moore.chrystal <@t> cryolife.com Fri Oct 10 14:46:55 2003 From: moore.chrystal <@t> cryolife.com (moore.chrystal@cryolife.com) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Histotechnologist Opening in NW Atlanta!! Message-ID: We currently have an opening for a Histotechnologist in NW Atlanta. Reporting to the Associate Medical Director, the main objective is to operate a *new* laboratory providing in-house histology services to support pathology and R&D. Responsibilities: Operation and maintenance of tissue processing equipment. Preparation of histologic slides, including routine staining, special histochemical staining, and immunohistochemistry (manual and automated.) Manage inventory of histology supplies. Assist pathologist in performing gross examination of pathology sections. Qualifications: Certified HT (ASCP) or equivalent. 2 yrs previous experience, supervisory level preferred. Skilled in routine histology, special histochemistry, and immunohistochemistry. Operation of histology equipment: tissue processor, embedding station, microtome, cryostat and automated stainer. Hours: 9 am to 6 pm Monday through Friday. CryoLife offers competitive salary and competitive benefits, which include: Health, Dental, Life, 401(k), Stock Purchase Plan, and Tuition Reimbursement. CryoLife's corporate headquarters are located in Kennesaw (NW Atlanta), GA in two buildings that total 200,000 square feet on a 21.5 acre, campus style setting. If you know if any candidates that may be interested, please have them contact me directly. Sincerely, Chrystal Moore, PHR Human Resources Coordinator CryoLife, Inc. Toll-Free: (800) 438-8285 Direct: (678) 290-4363 Fax: (770) 590-3741 Email: moore.chrystal@cryolife.com Website: www.cryolife.com CryoLife is an Equal Employment Opportunity Employer! Confidentiality Note: This e-mail message may contain information that is privileged and/or confidential. If you are not the addressee or an authorized recipient of this message, any distribution, copying, publication, or use of this information for any purpose is prohibited. Please notify the sender immediately by e-mail and then delete this message. From ohenry <@t> dfw.net Fri Oct 10 17:03:50 2003 From: ohenry <@t> dfw.net (Susan Owens) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] haze also silver's(SS) not staining Message-ID: <002401c38f7a$ae8a7f00$4fdd3040@Nationwide.net> I forgot to mention in my posts, on how we solved our 'haze' problem and the problem of the edges of the tissue not staining with Steiner/Steiner, our problems ONLY effected the GI's and all were first fixed in Hollands. The problems were never seen in the regular(non-Holland fixed ) tissue nor on any kind of tissue other then the GI's. Susan Owens-TX ohenry@dfw.net voice: 817-261-7938 fax: 817-548-9876 "A bad day at the dog show is better then a good day at work!" From stancelb <@t> msn.com Fri Oct 10 18:01:43 2003 From: stancelb <@t> msn.com (Barbara Stancel) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Shattering Mouse Brain Message-ID: Dear LaTricia, Although it sounds like a problem in processing, reprocessing may not be the answer you need right now. Your best bet at this time is to lower the temperature of the water bath in 5 degree increments until you get to a temperature where the tissues does not explode. Of course that may mean you have to leave the section on the water bath a few minutes to relax bumpy areas. Best of luck. Histologically yours, Barbara Barbara H. Stancel, HTL(ASCP)HT USDA, FSIS, OPHS, Eastern Laboratory, Pathology Athens, Georgia 30604 -----Original Message----- From: LaTricia Faison [mailto:lfaison@mail.mcg.edu] Sent: Wednesday, October 08, 2003 12:27 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Shattering Mouse Brain Does anyone know what may cause mouse brain to separate when put on the water bath? The tissue was given to me after it had been in paraformaldehyde for about 7-8 months. It was paraffin processed and shattered soon after touching the water bath. _________________________________________________________________ Instant message with integrated webcam using MSN Messenger 6.0. Try it now FREE! http://msnmessenger-download.com From dmccaig <@t> ckha.on.ca Fri Oct 10 18:29:39 2003 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] purple haze--?drying artifact Message-ID: I am curious to see if there is a relationship with the biopsies and wrapping them in some sort of filter paper. I have seen this drying effect if the biopsies are placed on dry paper and there is a delay in time wrapping them and getting them into the formalin. I moisten the paper with formalin prior to placing biopsies on them. Just another idea. Diana McCaig, R.T. Charge Tech, Histology Chatham Kent Health Alliance 519-352-6401 (3347) From hadi83 <@t> comcast.net Fri Oct 10 21:06:42 2003 From: hadi83 <@t> comcast.net (Hadi Yaziji) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] negative IHC controls In-Reply-To: Message-ID: <8EAC13A8-FB8F-11D7-81B9-00039378E76A@comcast.net> Theoretically, I agree with you that (a) running multiple negative control sections to cover the different antibodies/pretreatments is ideal and (b) money should be well spent on QC (I can't emphasize that enough). However, I honestly don't remember when was the last time when I had a false positive staining on a patient's slide that the negative control was the only way that allowed me to recognize the false positive signal on the real slide. And we look at 100s of IHC stains every day. Experienced pathologists and technologists can still recognize in > 99% of the times a false positive signal from a real signal on the tissue section of a given antibody without even having to look at negative controls. I don't think it should be a CAP requirement at all. In fact, I'd say the same thing on positive controls. All you need is one positive control per antibody, but not one control/per antibody/per patient. In many cases positive internal controls are present on the same slide, so you can tell whether your antibody worked simply by evaluating the expected positive internal controls. In fact, if your positive 'external' control worked and internal controls didn't, then you need to repeat your antibody test regardless of the positive external control. External controls are different tissues, fixed differently, processed differently and the tissue ages differently (in terms of its antigenicity). Individuals who perform and interpret IHC studies must have the knowledge to recognize the expected sub-cellular localization of every antibody on every type of tissue, including aberrant signals. For instance, you can easily dismiss a granular cytoplasmic TTF-1 signal as not positive, because TTF-1 is a nuclear transcription factor and we know it's expressed in the nuclei. However, hepatocellular carcinomas can give you a consistent and reproducible granular and cytoplasmic TTF-1 signal, and if you subsequently run confirmatory markers (HepPar1, polyclonal CEA, CD10, etc.) they will be positive. Such (granular cytoplasmic) signal should be recognized by the interpreter (tech or pathologist) as a specific signal for tumors with hepatoid phenotypes. This is just one of many examples.. The bottom line: in real life, one negative control per case is more than sufficient. And to say the least, it's inaccurate to state this is poor patient care and lousy quality control. I look forward to any constructive criticism. Hadi Yaziji, M.D. PhenoPath Laboratories On Thursday, October 9, 2003, at 07:03 AM, Horn, Hazel V wrote: > Not only should you be running a negative control for each patient > slide.?? That negative control should be treated just as your antibody > is.?? If the antibody is rabbit and antigen retrieved, so should your > control.?? If another antibody on the same patient is mouse and not > retrieved another negative control should be run with this same > protocol.?? In the United States, labs that are inspected by the CAP, > are required to run these controls.??? MONEY should never be > considered as a reason to stop doing a part of a procedure.?? It's > poor patient care and lousy quality control.?? IMHO. > ? > Hazel Horn, HT/HTL (ASCP) > Histology Supervisor > Arkansas Children's Hospital > > Phone - 501.364.4240 > Fax - 501.364.3912 > > > -----Original Message----- > From: vermast [mailto:vermast@rogers.com] > Sent: Wednesday, October 08, 2003 3:57 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] negative IHC controls > > ? > I would like to get a feel for how many out there are running negative > control slides for IHC.? > ? > In our lab we do just a handful of antibodies and initially I had been > running a negative control slide with each patient slide. ? After much > discussion with our pathologists, we decided to omit these negatives > (which were conistently negative)?and continue to just run a positive > control with each primary antibody for the run.? We use the Dako > autostainer and prediluted primaries.? The decision to stop running > negatives also coincided with Dako's decision to sell the negative > control sera separately from the primaries (they used to come packaged > together).? Perhaps I assumed that discontinuing to pair these > reagents together meant that few labs were using the negatives. > ? > Anyhow, after having reviewed the last QMPLS (Canada) survey committee > comments, I believe the committe would like a negative control run > with each patient tissue slide in order to evaluate background? (they > have used NCCLS guide pages as reference).? Incidentally we weren't a > part of the survey due to a technicality. > Any help or advice would be appreciated. > ? > L. Vermast > Stratford, Ont. > > > > > > > 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. Arkansas > Children's Hospital -------------- next part -------------- A non-text attachment was scrubbed... Name: not available Type: text/enriched Size: 5913 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031010/2547fa2f/attachment.bin From AliNeumann <@t> aol.com Fri Oct 10 22:03:54 2003 From: AliNeumann <@t> aol.com (AliNeumann@aol.com) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] STATIC AT THE MICROTOME Message-ID: <1c8.1031cd0e.2cb8cd1a@aol.com> I am learning to be a histotech and when using the manual Leitz 1512 microtome, I seem to have a problem with alot of static, the ribbons either move away from my fingers or they jump on and wrap around them, or they jump on to the disposable kinfe holder or the frame of the microtome. Does anyone have any suggestions they would be willing to share with me, to make my learning less irritating? Thank You, Bob Neumann -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031010/7dfe9e6f/attachment.htm From nick.kirk3 <@t> btopenworld.com Sat Oct 11 02:16:29 2003 From: nick.kirk3 <@t> btopenworld.com (Nick Kirk) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] negative IHC controls In-Reply-To: <8EAC13A8-FB8F-11D7-81B9-00039378E76A@comcast.net> Message-ID: A cautionary note I agree that false positives are rare but they do happen. We had a couple a while ago when it turned out that the Teflon coat on the probe of our DAKO Autostainer had perished and was carrying over reagents from one slide to another and also contaminating our negative control solution with primary antibody. Also they are very useful in checking that there is no endogenous biotin activity (more common than people think) and also that your hydrogen peroxide solution hasn't gone off when quenching endogenous peroxidase activity. As for the argument about only one positive control per antibody per run rather than per patient - Well the question I would pose is this. If in 5 years time you wish to review the immuno for a particular case, how do you ensure that the staining quality was adequate at the time if there aren't any positive controls for that case? On medico-legal grounds alone you should be doing positive controls for each case, especially if those results end up with the patient having a particularly aggressive treatment like chemotherapy or a particularly invasive surgical procedure such as a colectomy or a mastectomy. If someone wants to sue your organisation at a later date for inappropriate treatment you need every bit of proof that your part of the investigation was above reproach and controls is one thing that will definitely be asked about. I certainly wouldn't like to stand up in court and try and defend myself against that one! Incidentally, we use Surgipath's Control slides which allow you (most of the time depending on the size of the test section) to have the positive control and test section on the same slide, which saves a lot of space on the immunostainer and neatly solves the problem of where do you store the positive control if you use the single control per run model and you have multiple cases with the same antibody. It also acts as a check that the slide has received the correct primary antibody and that someone hasn't loaded the immunostainer wrongly. At the end of the day I still go by the analogy someone else made here earlier about airbags and seat belts in cars. Would you drive in a car without them? They may never be needed in your entire driving life, but you would be a dam fool not to have them wouldn't you? And I think we will have to agree to disagree Hadi, with your last statement, it is poor quality control by any definition of the term, not to use both negative and positive controls for each case. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Hadi Yaziji Sent: 11 October 2003 03:07 To: histonet@pathology.swmed.edu Subject: Re: [Histonet] negative IHC controls Theoretically, I agree with you that (a) running multiple negative control sections to cover the different antibodies/pretreatments is ideal and (b) money should be well spent on QC (I can't emphasize that enough). However, I honestly don't remember when was the last time when I had a false positive staining on a patient's slide that the negative control was the only way that allowed me to recognize the false positive signal on the real slide. And we look at 100s of IHC stains every day. Experienced pathologists and technologists can still recognize in > 99% of the times a false positive signal from a real signal on the tissue section of a given antibody without even having to look at negative controls. I don't think it should be a CAP requirement at all. In fact, I'd say the same thing on positive controls. All you need is one positive control per antibody, but not one control/per antibody/per patient. In many cases positive internal controls are present on the same slide, so you can tell whether your antibody worked simply by evaluating the expected positive internal controls. In fact, if your positive 'external' control worked and internal controls didn't, then you need to repeat your antibody test regardless of the positive external control. External controls are different tissues, fixed differently, processed differently and the tissue ages differently (in terms of its antigenicity). Individuals who perform and interpret IHC studies must have the knowledge to recognize the expected sub-cellular localization of every antibody on every type of tissue, including aberrant signals. For instance, you can easily dismiss a granular cytoplasmic TTF-1 signal as not positive, because TTF-1 is a nuclear transcription factor and we know it's expressed in the nuclei. However, hepatocellular carcinomas can give you a consistent and reproducible granular and cytoplasmic TTF-1 signal, and if you subsequently run confirmatory markers (HepPar1, polyclonal CEA, CD10, etc.) they will be positive. Such (granular cytoplasmic) signal should be recognized by the interpreter (tech or pathologist) as a specific signal for tumors with hepatoid phenotypes. This is just one of many examples.. The bottom line: in real life, one negative control per case is more than sufficient. And to say the least, it's inaccurate to state this is poor patient care and lousy quality control. I look forward to any constructive criticism. Hadi Yaziji, M.D. PhenoPath Laboratories On Thursday, October 9, 2003, at 07:03 AM, Horn, Hazel V wrote: Not only should you be running a negative control for each patient slide. That negative control should be treated just as your antibody is. If the antibody is rabbit and antigen retrieved, so should your control. If another antibody on the same patient is mouse and not retrieved another negative control should be run with this same protocol. In the United States, labs that are inspected by the CAP, are required to run these controls. MONEY should never be considered as a reason to stop doing a part of a procedure. It's poor patient care and lousy quality control. IMHO. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: vermast [mailto:vermast@rogers.com] Sent: Wednesday, October 08, 2003 3:57 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] negative IHC controls I would like to get a feel for how many out there are running negative control slides for IHC. In our lab we do just a handful of antibodies and initially I had been running a negative control slide with each patient slide. After much discussion with our pathologists, we decided to omit these negatives (which were conistently negative) and continue to just run a positive control with each primary antibody for the run. We use the Dako autostainer and prediluted primaries. The decision to stop running negatives also coincided with Dako's decision to sell the negative control sera separately from the primaries (they used to come packaged together). Perhaps I assumed that discontinuing to pair these reagents together meant that few labs were using the negatives. Anyhow, after having reviewed the last QMPLS (Canada) survey committee comments, I believe the committe would like a negative control run with each patient tissue slide in order to evaluate background (they have used NCCLS guide pages as reference). Incidentally we weren't a part of the survey due to a technicality. Any help or advice would be appreciated. L. Vermast Stratford, Ont. 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. Arkansas Children's Hospital -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031011/6632f2c4/attachment.htm From RCHIOVETTI <@t> aol.com Sat Oct 11 12:37:29 2003 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] STATIC AT THE MICROTOME Message-ID: <9d.3f7e0d04.2cb999d9@aol.com> Bob, Dry atmospheric conditions could be the culprit, also the clothes that you wear. Synthetic fabrics can cause real headaches. For dry conditions, some folks gently breathe on the block before sectioning; others dip their thumb in water and then gently rub their thumb on the block surface. Some folks use a small humidifier in the vicinity of the microtome. Others burn a small alcohol lamp somewhere nearby. Anything to increase the humidity in the vicinity of the block is worth a try. There is an interesting discussion in the Histonet Archives about this problem. The discussion thread mainly concerns static electricity in cryostats, and of course breathing on a frozen specimen is a no-no! But look for messages concerning "Breathers" and "Huffers" (the posters' choice of words, not mine!) These messages talk about static electricity and paraffin sections. For the Histonet community that might not know about it, the Histonet search engine is fantastic. I use it a lot. Here's the link: Click here: Histosearch: The Histology Search Engine (that's ). Just click on the box and select "Histonet Archives" as the database to search, enter your search words, and you'll get more info than you thought was out there. I used "static electricity" as the search words (without the quote marks). You would probably get more pertinent results by searching for "breathers" and "huffers." Good luck, hope this helps. Bob Chiovetti GTI Microsystems Leica Exclusive Regional Dealer Desert Southwest Region, USA From moore.chrystal <@t> cryolife.com Sat Oct 11 13:49:55 2003 From: moore.chrystal <@t> cryolife.com (moore.chrystal@cryolife.com) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Chrystal Moore is out of the office. Message-ID: I will be out of the office starting 10/11/2003 and will not return until 10/14/2003. I will be out of the office Monday, 10/13 and will return on Tuesday, 10/14. Sincerely, Chrystal Moore, PHR From georgecole <@t> ev1.net Sat Oct 11 14:35:10 2003 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Static charges while sectioning Message-ID: <000001c3902e$c9bd7d40$054dbad0@hppav> I'm surprised that none of you histotechs have heard of Static Master polonium emitters---I used them for years to quell sections flying on static winds, so to speak. The Static Master is harmless. Its emissions do not penetrate skin. The gadget comes with a warning not to swallow the metal under the grid---I've looked at that grid many times, wondering just how the heck anyone would go about swallowing those sharp little metal strips---that's not a dare----stick to goldfish if anyone of you wishes to swallow something weird! I have spent thousands of hours keeping static charges under control with these neat gadget---and I have no overt signs of any kind of reaction. I will go back to my old lab to look up the Static Master ordering address if none of you can find it. georgecole@ev1.net -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031011/316b149b/attachment.htm From lpwenk <@t> covad.net Sun Oct 12 05:48:20 2003 From: lpwenk <@t> covad.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] testing and new address Message-ID: <001f01c390ae$5a948d20$8732fea9@hppav> Had to unsubscribe, as our ISP has been bought out AGAIN (3rd, 4th time?). Finally, this newest company MADE us change our email address. So am now trying to re-subscribe to histonet. Please note, my home email address has changed Lpwenk@covad.net Have I missed much in the last 2 weeks? (rhetorical question. Don't expect an answer.) Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 From moore.chrystal <@t> cryolife.com Sun Oct 12 13:53:54 2003 From: moore.chrystal <@t> cryolife.com (moore.chrystal@cryolife.com) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Chrystal Moore is out of the office. Message-ID: I will be out of the office starting 10/11/2003 and will not return until 10/14/2003. I will be out of the office Monday, 10/13 and will return on Tuesday, 10/14. Sincerely, Chrystal Moore, PHR From mbecker <@t> pathlabinc.com Sun Oct 12 14:34:08 2003 From: mbecker <@t> pathlabinc.com (Michelle Becker) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Histotechs grossing? Message-ID: Hello Histo-world! Would anyone be willing to share the policy/ protocol they follow for allowing histotechs to examine and gross skin specimens? Histotechs have been grossing skins for 12 years in our independent lab. We were recently cited during a CAP inspection for not having such a policy in place. These gross examinations are also not supervised or evaluated by a pathologist since we do not have any on site. The slides for these specimens are directly returned to the dermatologist involved in the case. Are there any other labs out there with similar circumstances? If so, how do you handle this? Any help will be very much appreciated. michb From Stanley.Lupo <@t> gsk.com Mon Oct 13 10:54:56 2003 From: Stanley.Lupo <@t> gsk.com (Stanley.Lupo@gsk.com) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] online histology training Message-ID: Stanley.Lupo@GSK.Com Safety Assessment Mail Code: UE0461 Phone: 610 270-7340 Fax: 610 270-7202 Several people in my lab are studying for the exam. Here are the comments of one: You can tell this woman that there is a on line training course at www.hcconline.harford.edu The instructor is Floyd Grimm and it costs about 150.00. I am presently taking the course and it is very labor intensive. You pretty much write the definition of all the terms and stains in the Freida Carson book. I think it will help people pass the test because of the repetition but I think reading the book would be just as helpful. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031013/eccc5037/attachment.htm From histoclass03 <@t> hotmail.com Mon Oct 13 07:58:52 2003 From: histoclass03 <@t> hotmail.com (CAbetsy krummrey) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Ammoniacal Silver? Message-ID: Dear Histoneters: Is there a difference between ammoniacal and diamine silver?. Our Carsons workbook does not do a very good job at discribing the two silvers. histoclass03@hotmail.com _________________________________________________________________ Get MSN 8 Dial-up Internet Service FREE for one month. Limited time offer-- sign up now! http://join.msn.com/?page=dept/dialup From FARFAT <@t> aol.com Sat Oct 11 23:12:00 2003 From: FARFAT <@t> aol.com (FARFAT@aol.com) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] CD15 and CD3 Message-ID: <7a.494ff4b7.2cba2e90@aol.com> Hi Robert, If you want good stainging for the above 2 cds,Pls try cd-3 from Lab vision which is a rabbit mono and it is great. Secondly for CD-15 your link should be IGM and not IGG, i dont know what kind of dectection system you are using but for cd-15 get a link which is IGM and is available from Jackson labs. Good luck. Akbar. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031012/6969f971/attachment.htm From hadi83 <@t> comcast.net Sat Oct 11 10:21:37 2003 From: hadi83 <@t> comcast.net (Hadi Yaziji) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] negative IHC controls In-Reply-To: Message-ID: <9B27B2B0-FBFE-11D7-81B9-00039378E76A@comcast.net> Dear Nick, Thanks for the email. You definitely misread my email. First, where in my message did I advocate not to use negative and positive controls per case? Of course it's poor patient's care not to do that. Is it necessary to run + and - controls on every antibody? I don't think so provided you have the necessary experience to interpret them. To address your other point about reviewing cases, we keep our positive and negative controls indefinitely. You can always pull the controls that pertain to the run of the particular day of the case in question. You will be 'dam fool' (I am copying your words) to run IHC if you have no experience. That's much more dangerous legally and ethically. I've seen slides misinterpreted where the positive control is present on the same slide. Bottom line: one + control per run is adequate (provided you have the experience, which I'm not sure you have based on your reply). One - control per case is more than sufficient. I hope this addresses your concerns. Hadi Yaziji, M.D. PhenoPath Laboratories On Saturday, October 11, 2003, at 12:16 AM, Nick Kirk wrote: > A cautionary note > ? > I agree that false positives are rare but they do happen. We had a > couple a while ago when it turned out that the Teflon coat on the > probe of our DAKO Autostainer had perished and was carrying over > reagents from one slide to another and also contaminating our negative > control solution with primary antibody. > Also they are very useful in checking that there is no endogenous > biotin activity (more common than people think) and also that your > hydrogen peroxide solution hasn't gone off when quenching endogenous > peroxidase activity. > ? > As for the argument about only one positive control per antibody per > run rather than per patient - > Well the question I would pose is this. If in 5 years time you wish to > review the immuno for a particular case, how do you ensure that the > staining quality was adequate at the time if there aren't any positive > controls for that case? > On medico-legal grounds alone you should be doing positive controls > for each case, especially if those results end up with the patient > having a particularly aggressive treatment like chemotherapy or a > particularly invasive surgical procedure such as a colectomy or a > mastectomy. If someone wants to sue your organisation at a later date > for inappropriate treatment you need every bit of proof that your part > of the investigation was above reproach and controls is one thing that > will definitely be asked about. I certainly wouldn't like to stand up > in court and try and defend myself against that one! > ? > Incidentally, we use Surgipath's Control slides which allow you (most > of the time depending on the size of the test section) to have the > positive control and test section on the same slide, which saves a lot > of space on the immunostainer and neatly solves the problem of where > do you store the positive control if you use the single control per > run model and you have multiple cases with the same antibody. It also > acts as a check that the slide has received the correct primary > antibody and that someone hasn't loaded the immunostainer wrongly. > ? > At the end of the day I still go by the analogy someone else made here > earlier about airbags and seat belts in cars. Would you drive in a car > without them? They may never be needed in your entire driving life, > but you would be a dam fool not to have? them wouldn't you? > ? > And I?think we will have to agree to?disagree Hadi, with your last > statement, it is poor quality control by any definition of the term, > not to use both negative and positive controls for each case. > ? > Nick Kirk > Histopathology > Hinchingbrooke Hospital > Huntingdon > England > > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Hadi > Yaziji > Sent: 11 October 2003 03:07 > To: histonet@pathology.swmed.edu > Subject: Re: [Histonet] negative IHC controls > > Theoretically, I agree with you that (a) running multiple negative > control sections to cover the different antibodies/pretreatments is > ideal and (b) money should be well spent on QC (I can't emphasize that > enough). However, I honestly don't remember when was the last time > when I had a false positive staining on a patient's slide that the > negative control was the only way that allowed me to recognize the > false positive signal on the real slide. And we look at 100s of IHC > stains every day. > > Experienced pathologists and technologists can still recognize in > > 99% of the times a false positive signal from a real signal on the > tissue section of a given antibody without even having to look at > negative controls. I don't think it should be a CAP requirement at > all. In fact, I'd say the same thing on positive controls. All you > need is one positive control per antibody, but not one control/per > antibody/per patient. In many cases positive internal controls are > present on the same slide, so you can tell whether your antibody > worked simply by evaluating the expected positive internal controls. > In fact, if your positive 'external' control worked and internal > controls didn't, then you need to repeat your antibody test regardless > of the positive external control. External controls are different > tissues, fixed differently, processed differently and the tissue ages > differently (in terms of its antigenicity). > > Individuals who perform and interpret IHC studies must have the > knowledge to recognize the expected sub-cellular localization of every > antibody on every type of tissue, including aberrant signals. For > instance, you can easily dismiss a granular cytoplasmic TTF-1 signal > as not positive, because TTF-1 is a nuclear transcription factor and > we know it's expressed in the nuclei. However, hepatocellular > carcinomas can give you a consistent and reproducible granular and > cytoplasmic TTF-1 signal, and if you subsequently run confirmatory > markers (HepPar1, polyclonal CEA, CD10, etc.) they will be positive. > Such (granular cytoplasmic) signal should be recognized by the > interpreter (tech or pathologist) as a specific signal for tumors with > hepatoid phenotypes. This is just one of many examples.. > > The bottom line: in real life, one negative control per case is more > than sufficient. And to say the least, it's inaccurate to state this > is poor patient care and lousy quality control. > > I look forward to any constructive criticism. > > Hadi Yaziji, M.D. > PhenoPath Laboratories > > On Thursday, October 9, 2003, at 07:03 AM, Horn, Hazel V wrote: > > Not only should you be running a negative control for each patient > slide.?? That negative control should be treated just as your antibody > is.?? If the antibody is rabbit and antigen retrieved, so should your > control.?? If another antibody on the same patient is mouse and not > retrieved another negative control should be run with this same > protocol.?? In the United States, labs that are inspected by the CAP, > are required to run these controls.??? MONEY should never be > considered as a reason to stop doing a part of a procedure.?? It's > poor patient care and lousy quality control.?? IMHO. > ? > Hazel Horn, HT/HTL (ASCP) > Histology Supervisor > Arkansas Children's Hospital > > Phone - 501.364.4240 > Fax - 501.364.3912 > > > -----Original Message----- > From: vermast [mailto:vermast@rogers.com] > Sent: Wednesday, October 08, 2003 3:57 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] negative IHC controls > > ? > I would like to get a feel for how many out there are running negative > control slides for IHC.? > ? > In our lab we do just a handful of antibodies and initially I had been > running a negative control slide with each patient slide. ? After much > discussion with our pathologists, we decided to omit these negatives > (which were conistently negative)?and continue to just run a positive > control with each primary antibody for the run.? We use the Dako > autostainer and prediluted primaries.? The decision to stop running > negatives also coincided with Dako's decision to sell the negative > control sera separately from the primaries (they used to come packaged > together).? Perhaps I assumed that discontinuing to pair these > reagents together meant that few labs were using the negatives. > ? > Anyhow, after having reviewed the last QMPLS (Canada) survey committee > comments, I believe the committe would like a negative control run > with each patient tissue slide in order to evaluate background? (they > have used NCCLS guide pages as reference).? Incidentally we weren't a > part of the survey due to a technicality. > Any help or advice would be appreciated. > ? > L. Vermast > Stratford, Ont. > > > > > > > > > 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. Arkansas > Children's Hospital > -------------- next part -------------- A non-text attachment was scrubbed... Name: not available Type: text/enriched Size: 10951 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031011/b682fb6e/attachment.bin From ploykasek <@t> phenopath.com Mon Oct 13 10:51:56 2003 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] IHC QC's Message-ID: I'm glad that everyone is so concerned with both negative and positive IHC controls. There is certainly more than one side to this issue. I will say that I don't think a positive QC on every slide is absolutely necessary, for many reasons. If the QC is rare & precious, then it is a waste of resources. As is running a negative control for every possible technique permutation on small amounts of tumor. I would rather have slides with tumor left for additional studies than have wasted tumor sections on 4-6 negative controls. You can always evaluate non-specific staining on slides that have had an antibody applied & that are negative with that antibody. The CAP is specific that positive controls be used for each antibody - see CAP checklist ANP.22550. They do not specify for each slide. Since positive QC's should be kept filed for the same number of years as the patient slide & records, it should be possible to pull a QC slide from the IHC run for a particular slide. In the CAP comment on ANP.22550, the use of internal QC's is also mentioned. Although there are many ways of dealing with the issue of QC's, I'm sure we all want to do what is prudent, abide by the regulations, and increase the level of patient care. Just my 2 cents worth. Patti Loykasek Phenopath Laboratories Seattle, WA -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031013/7f7bc28d/attachment.htm From Marion.Hiles <@t> north-bristol.swest.nhs.uk Mon Oct 13 08:59:21 2003 From: Marion.Hiles <@t> north-bristol.swest.nhs.uk (Marion Hiles) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Cyto Preps Message-ID: <533E4A1C3061FC4EB388119D2FD7B5D40243180B@nbfexch04.north-bristol.nhs> There was a good article,written by some guys at the Royal Victoria Hospital, Belfast, Ireland on 'Cytology preparation for immuno' for bloody/proteinaceous fluids in the DakoFacts publication Vol 02 No.01. Contact DakoCytomation. Bob Quilty Neuropathology Frenchay Hospital Bristol UK -----Original Message----- From: Cindy/Rick DuBois [mailto:ccdub@earthlink.net] Sent: 10 October 2003 17:43 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Cyto Preps We are interested in what reagents/procedures other labs are using to process bloody body fluids. What are you using to lyse the RBC's? We currently use plasma/thrombin, but our cytotechs are having a difficult time reading the slides. Does anyone know of a rapid method that does not interfere with immuno stains? Cindy DuBois, HT DELTA PATHOLOGY ASSOC. STOCKTON CA. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JLE <@t> rice.willmar.mn.us Mon Oct 13 13:09:13 2003 From: JLE <@t> rice.willmar.mn.us (Jennifer Englin) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] prostate needle biopsies Message-ID: We usually receive our needle biopsies in 2 contaners, right and left. I was wondering how many cassettes they are submitted in. For example 2 needles per cassette, five per cassette, etc... Also for those of you taking sections for possible IP's. We routinely cut five levels, 1,3,&5 for H&E and 2 & 4 for IP's. Do some of you cut seven??? Any help would be appreciated. Jennifer Englin, HT(ASCP),PA Rice Memorial Hospital Willmar, MN From jmacdona <@t> mtsac.edu Mon Oct 13 13:24:46 2003 From: jmacdona <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Need silver stain for granules in adrenals References: <3F85CFC8.9020308@umdnj.edu> Message-ID: <001501c391b7$486299d0$7934908c@JMACDONADT> According to Carson, the demonstration of chromaffin granules canot be demonstrated after formalin fixation. (Histotechnology: A Self-Instructional Text, page 6-7) Jennifer MacDonald ----- Original Message ----- From: Geoff McAuliffe To: Nick Kirk Cc: Histonet Sent: Thursday, October 09, 2003 2:14 PM Subject: Re: [Histonet] Need silver stain for granules in adrenals I don't think you can get a good chromaffin reaction on material already fixed with formalin. Fontana-Masson should be fine. John Kiernan published a chromate-dichromate fix for chromaffin tissues, it may be in his book, along with other recommendations. Geoff Nick Kirk wrote: What you are trying to demonstrate are Chromaffin cells of the adrenal glands so a Masson-Fontana should work, if not try a Schmorl or carry out the Chromaffin reaction If you use Iodate oxidation you can distinguish between Adrenaline and noradrenalin fairly easily as iodates oxidise noradrenaline much quicker than they do adrenaline. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Nancy Maronto Sent: 09 October 2003 16:05 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Need silver stain for granules in adrenals Hi, Could anyone suggest a stain for noradrenalin and adrenalin granules in paraffin embedded adrenals. We were asked to do a gremilius stain and it did not stain the granules. Are these slides lost or can we re stain them? We usually do a recut of the block, but it is asked if we could use the same tissue slide. The stain actually worked, but not for the granules. From what we read a Fontana-masson should work. There probably is a better stain our there for this specific target. Can anyone with some experience with staining these granules share their staining protocol. At this time the request is not for immuno staining. Thanks for your help. Nancy Maronto -- -- ********************************************** 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 ********************************************** -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031013/75e2e4fe/attachment.htm From stancelb <@t> msn.com Mon Oct 13 13:24:54 2003 From: stancelb <@t> msn.com (Barbara Stancel) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] STATIC AT THE MICROTOME Message-ID: Dear Bob, Dryer sheets. Try them in your pockets or stuffed up your sleeves. Rub them on your hands as you would toweling off after washing. I have even used them wrapped around the block (chuck) holder of the microtome or stuffed in the paraffin tray under the knive. They come fragranced and non-fragranced. I have also tried the anti-static sprays sold for spraying on clothes to reduce "creeping" skirts and pants. Works pretty good sprayed on microtomes and/or sleeves. There have been dry winters that I have used all of the above. Histologically yours, Barbara Barbara H. Stancel, HTL(ASCP)HT USDA, FSIS, OPHS, Eastern Laboratory, Pathology RRC, 950 College Station Road Athens, Georgia >From: AliNeumann@aol.com >To: histonet@pathology.swmed.edu >Subject: [Histonet] STATIC AT THE MICROTOME >Date: Fri, 10 Oct 2003 23:03:54 EDT > >I am learning to be a histotech and when using the manual Leitz 1512 >microtome, I seem to have a problem with alot of static, the ribbons either >move away >from my fingers or they jump on and wrap around them, or they jump on to >the >disposable kinfe holder or the frame of the microtome. Does anyone have any >suggestions they would be willing to share with me, to make my learning >less >irritating? >Thank You, >Bob Neumann _________________________________________________________________ Frustrated with dial-up? Get high-speed for as low as $29.95/month (depending on the local service providers in your area). https://broadband.msn.com From garygill <@t> dcla.com Mon Oct 13 13:29:04 2003 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Cyto Preps Message-ID: Specific 24-page issue: http://www.dakocytomation.com/dakofacts_4_final.pdf DakoFacts archives: http://www.dakocytomation.us/index/press_publications-2.htm Gary Gill -----Original Message----- From: Marion Hiles [mailto:Marion.Hiles@north-bristol.swest.nhs.uk] Sent: Monday, October 13, 2003 8:59 AM To: 'Cindy/Rick DuBois' Cc: Histonet (E-mail) Subject: RE: [Histonet] Cyto Preps There was a good article,written by some guys at the Royal Victoria Hospital, Belfast, Ireland on 'Cytology preparation for immuno' for bloody/proteinaceous fluids in the DakoFacts publication Vol 02 No.01. Contact DakoCytomation. Bob Quilty Neuropathology Frenchay Hospital Bristol UK -----Original Message----- From: Cindy/Rick DuBois [mailto:ccdub@earthlink.net] Sent: 10 October 2003 17:43 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Cyto Preps We are interested in what reagents/procedures other labs are using to process bloody body fluids. What are you using to lyse the RBC's? We currently use plasma/thrombin, but our cytotechs are having a difficult time reading the slides. Does anyone know of a rapid method that does not interfere with immuno stains? Cindy DuBois, HT DELTA PATHOLOGY ASSOC. STOCKTON CA. _______________________________________________ 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 cdemarinis <@t> SARATOGACARE.ORG Mon Oct 13 13:53:00 2003 From: cdemarinis <@t> SARATOGACARE.ORG (Demarinis,Carolyn) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] Gimenez technique Message-ID: <2FCF6059F121BA41A672C5012BB113AA2A52B0@mercury.saratogacare.org> We were having problems with the Gimenez stain for H. Pylori and I previously asked for help on the Histonet. Fortunately, Polyscientific staff contacted me and helped me troubleshoot the problem. The Gimenez stain is now working great and we are using it once again. CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please notify Saratoga Hospital immediately by e-mail at privacy@saratogacare.org and destroy all copies of this communication and any attachments. From laurie.colbert <@t> huntingtonhospital.com Mon Oct 13 14:33:27 2003 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] prostate needle biopsies Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5BC19@EXCHANGE1.huntingtonhospital.com> Jennifer, Our pathologists will submit as many prostate needle cores in one cassette that were received in the container. We generally don't receive more than three cores in one container, though. Also, we cut six levels: 1, 3, 5 for H&E and 2, 4, 6 for possible IP's. And, we routinely get six specimens per patient - yes, we waste a lot of slides:) Laurie Colbert Huntington Hospital Pasadena, CA -----Original Message----- From: Jennifer Englin [mailto:JLE@rice.willmar.mn.us] Sent: Monday, October 13, 2003 11:09 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] prostate needle biopsies We usually receive our needle biopsies in 2 contaners, right and left. I was wondering how many cassettes they are submitted in. For example 2 needles per cassette, five per cassette, etc... Also for those of you taking sections for possible IP's. We routinely cut five levels, 1,3,&5 for H&E and 2 & 4 for IP's. Do some of you cut seven??? Any help would be appreciated. Jennifer Englin, HT(ASCP),PA Rice Memorial Hospital Willmar, MN _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From HornHV <@t> archildrens.org Mon Oct 13 15:02:04 2003 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] IHC QC's Message-ID: This is from the March 2003 checklist............................. ANP.22570 Phase II N/A YES NO Are negative controls used for each antibody species? NOTE: A negative control for each primary antibody species must be used. Alternatively, buffer controls can be used if multiple antibodies for each species are included. The controls used should also control for pre-treatment conditions. COMMENTARY: A negative control for each primary antibody species must be used. Alternatively, buffer controls can be used if multiple antibodies for each species are included. The controls used should also control for pre-treatment conditions. REFERENCE: Weirauch M. Multitissue control block for immunohistochemistry. Lab Med. 1999;30:448-449. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: Patti Loykasek [mailto:ploykasek@phenopath.com] Sent: Monday, October 13, 2003 10:52 AM To: histonet Subject: [Histonet] IHC QC's I'm glad that everyone is so concerned with both negative and positive IHC controls. There is certainly more than one side to this issue. I will say that I don't think a positive QC on every slide is absolutely necessary, for many reasons. If the QC is rare & precious, then it is a waste of resources. As is running a negative control for every possible technique permutation on small amounts of tumor. I would rather have slides with tumor left for additional studies than have wasted tumor sections on 4-6 negative controls. You can always evaluate non-specific staining on slides that have had an antibody applied & that are negative with that antibody. The CAP is specific that positive controls be used for each antibody - see CAP checklist ANP.22550. They do not specify for each slide. Since positive QC's should be kept filed for the same number of years as the patient slide & records, it should be possible to pull a QC slide from the IHC run for a particular slide. In the CAP comment on ANP.22550, the use of internal QC's is also mentioned. Although there are many ways of dealing with the issue of QC's, I'm sure we all want to do what is prudent, abide by the regulations, and increase the level of patient care. Just my 2 cents worth. Patti Loykasek Phenopath Laboratories Seattle, WA 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. Arkansas Children's Hospital -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031013/95797e01/attachment.htm From HornHV <@t> archildrens.org Mon Oct 13 15:06:00 2003 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] IHC QC's Message-ID: I don't think anyone said a POSTIVE control should be run with each slide. We were talking about negative controls, I believe. I just copied and pasted from the latest CAP survey in another email. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: Patti Loykasek [mailto:ploykasek@phenopath.com] Sent: Monday, October 13, 2003 10:52 AM To: histonet Subject: [Histonet] IHC QC's I'm glad that everyone is so concerned with both negative and positive IHC controls. There is certainly more than one side to this issue. I will say that I don't think a positive QC on every slide is absolutely necessary, for many reasons. If the QC is rare & precious, then it is a waste of resources. As is running a negative control for every possible technique permutation on small amounts of tumor. I would rather have slides with tumor left for additional studies than have wasted tumor sections on 4-6 negative controls. You can always evaluate non-specific staining on slides that have had an antibody applied & that are negative with that antibody. The CAP is specific that positive controls be used for each antibody - see CAP checklist ANP.22550. They do not specify for each slide. Since positive QC's should be kept filed for the same number of years as the patient slide & records, it should be possible to pull a QC slide from the IHC run for a particular slide. In the CAP comment on ANP.22550, the use of internal QC's is also mentioned. Although there are many ways of dealing with the issue of QC's, I'm sure we all want to do what is prudent, abide by the regulations, and increase the level of patient care. Just my 2 cents worth. Patti Loykasek Phenopath Laboratories Seattle, WA 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. Arkansas Children's Hospital -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031013/bd6b67d1/attachment.htm From bhewlett <@t> cogeco.ca Mon Oct 13 15:26:35 2003 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] RE: negative IHC controls Message-ID: <004b01c391c8$4cb4f440$6400a8c0@bryaniwx13voft> Dear Hadi, I agree with much of the content of your posting. However, there seems to be a mixed message here. You agree that; (a) running multiple reagent control sections covering the appropriate antibodies/pretreatments is ideal and that, (b) money should be well spent on QC (which you emphasize), yet you seem to downplay the use of complete reagent controls and to inject some confusion regarding the necessary use of process controls. Process controls are intended to address the full range of procedural variables and to ensure that the end result of the process is known to lie within legitimate or acceptable values and is valid. Process controls are a necessary tool utilized PRIOR to the subsequent interpretation of the result. Subsequent interpretation of the result depends largely upon the validity provided by these process controls. No one has suggested that a reagent control on the patient slide is the only way to recognize a false positive signal. In fact, the experienced observer provides the final additional level of process control. However, that does not mean that all the necessary prior process controls can be avoided or omitted by that experienced observer. We too, have seen 100's of IHC stains daily and the full use of process controls has been an important factor in obtaining consistent results. We have seen numerous occasions where reagent controls have provided essential clues to the nature of an aberrant finding. I would have to disagree with you that a single reagent control is more than sufficient. I do agree with your comments regarding the use of one positive 'external' tissue control per antibody/ per run but would point out that, if the positive 'external' control works and the patient 'expected' positive internal controls didn't, repeating the antibody test, using the same process, should produce exactly the same result! That would mean that either the 'expected' result is not a valid expectation, or that the process controls are inadequate and do not account for all of the procedural variables. Regards, Bryan Hewlett On Friday, October 10, 2003, at 10.06 PM, Hadi Yaziji wrote: Theoretically, I agree with you that (a) running multiple negative control sections to cover the different antibodies/pretreatments is ideal and (b) money should be well spent on QC (I can't emphasize that enough). However, I honestly don't remember when was the last time when I had a false positive staining on a patient's slide that the negative control was the only way that allowed me to recognize the false positive signal on the real slide. And we look at 100s of IHC stains every day. ........ Experienced pathologists and technologists can still recognize in > 99% of the times a false positive signal from a real signal on the tissue section of a given antibody without even having to look at negative controls. ... Individuals who perform and interpret IHC studies must have the knowledge to recognize the expected sub-cellular localization of every antibody on every type of tissue, including aberrant signals.... All you need is one positive control per antibody, but not one control/per antibody/per patient. In many cases positive internal controls are present on the same slide, so you can tell whether your antibody worked simply by evaluating the expected positive internal controls. In fact, if your positive 'external' control worked and internal controls didn't, then you need to repeat your antibody test regardless of the positive external control........ The bottom line: in real life, one negative control per case is more than sufficient. And to say the least, it's inaccurate to state this is poor patient care and lousy quality control. From rrichar3 <@t> iupui.edu Mon Oct 13 15:41:23 2003 From: rrichar3 <@t> iupui.edu (Richardson, Rose M) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] paraffin processing for placenta Message-ID: <9CBE06ABD51D4345BAC9BE3C8D2E7671AC11D2@iu-mssg-mbx04.exchange.iu.edu> We are going to be processing human placenta for a special research project and need some ideas about timing for the tissue processor. Any tips would be greatly appreciated. Rose Richardson (Dr. Ghetti's Laboratory) Ind.Univ.Med.Sch./Pathology 635 Barnhill Drive Room B029 Indianapolis, IN 46202 317-274-1591 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031013/fa713a8c/attachment.htm From ploykasek <@t> phenopath.com Mon Oct 13 16:18:00 2003 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] IHC QC's In-Reply-To: Message-ID: There was in fact, a post from Nick Kirk on running a positive control with each case. I do realize the CAP requirements and am familiar with the checklist. Patti Loykasek Phenopath Laboratories Seattle, WA > > > I don't think anyone said a POSTIVE control should be run with each slide. > We were talking about negative controls, I believe. > I just copied and pasted from the latest CAP survey in another email. > > > Hazel Horn, HT/HTL (ASCP) > Histology Supervisor > Arkansas Children's Hospital > > Phone - 501.364.4240 > Fax - 501.364.3912 >> >> -----Original Message----- >> From: Patti Loykasek [mailto:ploykasek@phenopath.com] >> Sent: Monday, October 13, 2003 10:52 AM >> To: histonet >> Subject: [Histonet] IHC QC's >> >> I'm glad that everyone is so concerned with both negative and positive IHC >> controls. There is certainly more than one side to this issue. I will say >> that I don't think a positive QC on every slide is absolutely necessary, for >> many reasons. If the QC is rare & precious, then it is a waste of resources. >> As is running a negative control for every possible technique permutation on >> small amounts of tumor. I would rather have slides with tumor left for >> additional studies than have wasted tumor sections on 4-6 negative controls. >> You can always evaluate non-specific staining on slides that have had an >> antibody applied & that are negative with that antibody. The CAP is specific >> that positive controls be used for each antibody - see CAP checklist >> ANP.22550. They do not specify for each slide. Since positive QC's should be >> kept filed for the same number of years as the patient slide & records, it >> should be possible to pull a QC slide from the IHC run for a particular >> slide. In the CAP comment on ANP.22550, the use of internal QC's is also >> mentioned. Although there are many ways of dealing with the issue of QC's, >> I'm sure we all want to do what is prudent, abide by the regulations, and >> increase the level of patient care. >> Just my 2 cents worth. >> >> Patti Loykasek >> Phenopath Laboratories >> Seattle, WA > > 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. Arkansas Children's Hospital > -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031013/c8745f80/attachment.htm From nick.kirk3 <@t> btopenworld.com Mon Oct 13 17:00:50 2003 From: nick.kirk3 <@t> btopenworld.com (Nick Kirk) Date: Fri Sep 16 15:22:02 2005 Subject: [Histonet] RE: negative IHC controls In-Reply-To: <9B27B2B0-FBFE-11D7-81B9-00039378E76A@comcast.net> Message-ID: Hadi I am sorry but I must take issue with your very condescending and if I may say so insulting tone. For your information, I have over 18 years of experience doing IHC and my lab does a considerable number of slides per day. I have set up the IHC service in two labs and have received much praise for doing it. I have also done extensive lab based trials work for both DAKO and Novocastra here in the UK as well, so I think my credentials are better than most. I'm not at issue with the interpretation issue at all, that is indeed a very skilled task that takes many years of study and experience to become competent in. What I am taking issue with is the issues around clinical audit and best practice. Now I don't know what clinical audit systems you have in your establishment, but in ours it is regarded as poor clinical governance i.e. bad practice by both myself, the Head Biomedical Scientist of the department and all of my Pathologist colleagues not to have separate controls per case. This is especially the case when you are reviewing tumour antigenicity on samples that may come out of the archive. All our slides are stored in case order so to have positive controls for each case actually makes it much easier the extract all the relevant slides per case for later review. Having slides for the same case filed in separate places introduces the potential for error, especially when being initially filed. I know this to be true because it has happened to me on several occasions where slides filed in different places have been mis-filed and become difficult to retrieve. In the UK we use a system of evidence based good practice and evidence suggests that is precisely what we are doing. I hope that now explains it to you. Nick Kirk BSc (Hons) FIBMS (I've got letters too) Head Biomedical Scientist Histopathology Hinchingbrooke Hospital Huntingdon Cambridgeshire England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Hadi Yaziji Sent: 11 October 2003 16:22 To: histonet@pathology.swmed.edu Subject: Re: [Histonet] negative IHC controls Dear Nick, Thanks for the email. You definitely misread my email. First, where in my message did I advocate not to use negative and positive controls per case? Of course it's poor patient's care not to do that. Is it necessary to run + and - controls on every antibody? I don't think so provided you have the necessary experience to interpret them. To address your other point about reviewing cases, we keep our positive and negative controls indefinitely. You can always pull the controls that pertain to the run of the particular day of the case in question. You will be 'dam fool' (I am copying your words) to run IHC if you have no experience. That's much more dangerous legally and ethically. I've seen slides misinterpreted where the positive control is present on the same slide. Bottom line: one + control per run is adequate (provided you have the experience, which I'm not sure you have based on your reply). One - control per case is more than sufficient. I hope this addresses your concerns. Hadi Yaziji, M.D. PhenoPath Laboratories On Saturday, October 11, 2003, at 12:16 AM, Nick Kirk wrote: A cautionary note I agree that false positives are rare but they do happen. We had a couple a while ago when it turned out that the Teflon coat on the probe of our DAKO Autostainer had perished and was carrying over reagents from one slide to another and also contaminating our negative control solution with primary antibody. Also they are very useful in checking that there is no endogenous biotin activity (more common than people think) and also that your hydrogen peroxide solution hasn't gone off when quenching endogenous peroxidase activity. As for the argument about only one positive control per antibody per run rather than per patient - Well the question I would pose is this. If in 5 years time you wish to review the immuno for a particular case, how do you ensure that the staining quality was adequate at the time if there aren't any positive controls for that case? On medico-legal grounds alone you should be doing positive controls for each case, especially if those results end up with the patient having a particularly aggressive treatment like chemotherapy or a particularly invasive surgical procedure such as a colectomy or a mastectomy. If someone wants to sue your organisation at a later date for inappropriate treatment you need every bit of proof that your part of the investigation was above reproach and controls is one thing that will definitely be asked about. I certainly wouldn't like to stand up in court and try and defend myself against that one! Incidentally, we use Surgipath's Control slides which allow you (most of the time depending on the size of the test section) to have the positive control and test section on the same slide, which saves a lot of space on the immunostainer and neatly solves the problem of where do you store the positive control if you use the single control per run model and you have multiple cases with the same antibody. It also acts as a check that the slide has received the correct primary antibody and that someone hasn't loaded the immunostainer wrongly. At the end of the day I still go by the analogy someone else made here earlier about airbags and seat belts in cars. Would you drive in a car without them? They may never be needed in your entire driving life, but you would be a dam fool not to have them wouldn't you? And I think we will have to agree to disagree Hadi, with your last statement, it is poor quality control by any definition of the term, not to use both negative and positive controls for each case. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Hadi Yaziji Sent: 11 October 2003 03:07 To: histonet@pathology.swmed.edu Subject: Re: [Histonet] negative IHC controls Theoretically, I agree with you that (a) running multiple negative control sections to cover the different antibodies/pretreatments is ideal and (b) money should be well spent on QC (I can't emphasize that enough). However, I honestly don't remember when was the last time when I had a false positive staining on a patient's slide that the negative control was the only way that allowed me to recognize the false positive signal on the real slide. And we look at 100s of IHC stains every day. Experienced pathologists and technologists can still recognize in > 99% of the times a false positive signal from a real signal on the tissue section of a given antibody without even having to look at negative controls. I don't think it should be a CAP requirement at all. In fact, I'd say the same thing on positive controls. All you need is one positive control per antibody, but not one control/per antibody/per patient. In many cases positive internal controls are present on the same slide, so you can tell whether your antibody worked simply by evaluating the expected positive internal controls. In fact, if your positive 'external' control worked and internal controls didn't, then you need to repeat your antibody test regardless of the positive external control. External controls are different tissues, fixed differently, processed differently and the tissue ages differently (in terms of its antigenicity). Individuals who perform and interpret IHC studies must have the knowledge to recognize the expected sub-cellular localization of every antibody on every type of tissue, including aberrant signals. For instance, you can easily dismiss a granular cytoplasmic TTF-1 signal as not positive, because TTF-1 is a nuclear transcription factor and we know it's expressed in the nuclei. However, hepatocellular carcinomas can give you a consistent and reproducible granular and cytoplasmic TTF-1 signal, and if you subsequently run confirmatory markers (HepPar1, polyclonal CEA, CD10, etc.) they will be positive. Such (granular cytoplasmic) signal should be recognized by the interpreter (tech or pathologist) as a specific signal for tumors with hepatoid phenotypes. This is just one of many examples.. The bottom line: in real life, one negative control per case is more than sufficient. And to say the least, it's inaccurate to state this is poor patient care and lousy quality control. I look forward to any constructive criticism. Hadi Yaziji, M.D. PhenoPath Laboratories On Thursday, October 9, 2003, at 07:03 AM, Horn, Hazel V wrote: Not only should you be running a negative control for each patient slide. That negative control should be treated just as your antibody is. If the antibody is rabbit and antigen retrieved, so should your control. If another antibody on the same patient is mouse and not retrieved another negative control should be run with this same protocol. In the United States, labs that are inspected by the CAP, are required to run these controls. MONEY should never be considered as a reason to stop doing a part of a procedure. It's poor patient care and lousy quality control. IMHO. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: vermast [mailto:vermast@rogers.com] Sent: Wednesday, October 08, 2003 3:57 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] negative IHC controls I would like to get a feel for how many out there are running negative control slides for IHC. In our lab we do just a handful of antibodies and initially I had been running a negative control slide with each patient slide. After much discussion with our pathologists, we decided to omit these negatives (which were conistently negative) and continue to just run a positive control with each primary antibody for the run. We use the Dako autostainer and prediluted primaries. The decision to stop running negatives also coincided with Dako's decision to sell the negative control sera separately from the primaries (they used to come packaged together). Perhaps I assumed that discontinuing to pair these reagents together meant that few labs were using the negatives. Anyhow, after having reviewed the last QMPLS (Canada) survey committee comments, I believe the committe would like a negative control run with each patient tissue slide in order to evaluate background (they have used NCCLS guide pages as reference). Incidentally we weren't a part of the survey due to a technicality. Any help or advice would be appreciated. L. Vermast Stratford, Ont. 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. Arkansas Children's Hospital -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031013/981c9fe4/attachment.htm From scott.turner <@t> dnax.org Mon Oct 13 18:57:23 2003 From: scott.turner <@t> dnax.org (Turner, Scott) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Jones/PAS Message-ID: <29B25753F6B1D51196110002A589D4444EC06C@PALMSG30.us.schp.com> A colleague of mine needs to do a Jones' silver stain in kidney in combination with PAS. Does anyone have a protocol for doing these two together? Scott Turner DNAX Research Institute Palo Alto, CA ********************************************************************* 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. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031013/7a323027/attachment.htm From jnocito <@t> satx.rr.com Mon Oct 13 21:48:16 2003 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] IHC QC's References: Message-ID: <003901c391fd$a2917160$70494542@satx.rr.com> Re: [Histonet] IHC QC'sI've always run one positive control for each antibody and a negative control for each paraffin block. When I was the immuno supervisor at AFIP (during another life) we would anywhere from 25-100 cases of the same antibody, i.e. we would run 1 CD45 control and 50 patient slides, but each patient slide would have 1 negative. I haven't had a problem yet, including both the CAP and CLIA inspections I went through this year. I have to agree with Patti, since I work in a reference lab, we don't receive the good cases that a hospital would so we have to make do with what we can get. Joe Nocito BS, HT (ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, Texas ----- Original Message ----- From: Patti Loykasek To: Horn, Hazel V ; histonet Sent: Monday, October 13, 2003 2:18 PM Subject: Re: [Histonet] IHC QC's There was in fact, a post from Nick Kirk on running a positive control with each case. I do realize the CAP requirements and am familiar with the checklist. Patti Loykasek Phenopath Laboratories Seattle, WA I don't think anyone said a POSTIVE control should be run with each slide. We were talking about negative controls, I believe. I just copied and pasted from the latest CAP survey in another email. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: Patti Loykasek [mailto:ploykasek@phenopath.com] Sent: Monday, October 13, 2003 10:52 AM To: histonet Subject: [Histonet] IHC QC's I'm glad that everyone is so concerned with both negative and positive IHC controls. There is certainly more than one side to this issue. I will say that I don't think a positive QC on every slide is absolutely necessary, for many reasons. If the QC is rare & precious, then it is a waste of resources. As is running a negative control for every possible technique permutation on small amounts of tumor. I would rather have slides with tumor left for additional studies than have wasted tumor sections on 4-6 negative controls. You can always evaluate non-specific staining on slides that have had an antibody applied & that are negative with that antibody. The CAP is specific that positive controls be used for each antibody - see CAP checklist ANP.22550. They do not specify for each slide. Since positive QC's should be kept filed for the same number of years as the patient slide & records, it should be possible to pull a QC slide from the IHC run for a particular slide. In the CAP comment on ANP.22550, the use of internal QC's is also mentioned. Although there are many ways of dealing with the issue of QC's, I'm sure we all want to do what is prudent, abide by the regulations, and increase the level of patient care. Just my 2 cents worth. Patti Loykasek Phenopath Laboratories Seattle, WA ---------------------------------------------------------------------------- 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. Arkansas Children's Hospital -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031013/af67397c/attachment.htm From bhewlett <@t> cogeco.ca Mon Oct 13 19:53:55 2003 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Jones/PAS References: <29B25753F6B1D51196110002A589D4444EC06C@PALMSG30.us.schp.com> Message-ID: <000801c391ed$a571ef50$6400a8c0@bryaniwx13voft> Jones/PASScott, It would be pointless to perform a PAS and a Jones silver together as a combination since they both demonstrate the same elements. Jones silver method for basement membranes, uses the methenamine silver as an aldehyde detection system in place of the Schiff reagent used in PAS. The reason for this is to provide a more intense and contrasty demonstration of the basement membrane in thin sections of kidney. A PAS stain tends to be too weak in 1-2 micrometer sections. Regards, Bryan ----- Original Message ----- From: Turner, Scott To: HistoNet Server (histonet@pathology.swmed.edu) Sent: Monday, October 13, 2003 7:57 PM Subject: [Histonet] Jones/PAS A colleague of mine needs to do a Jones' silver stain in kidney in combination with PAS. Does anyone have a protocol for doing these two together? Scott Turner DNAX Research Institute Palo Alto, CA ********************************************************************* 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. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031013/d56990b9/attachment.htm From jnocito <@t> satx.rr.com Mon Oct 13 21:54:00 2003 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] prostate needle biopsies References: Message-ID: <005001c391fe$707d81e0$70494542@satx.rr.com> Jennifer, we usually get one, maybe two cores per container, but we get anywhere from 8 to 16 containers per case. We cut 3 levels, on 3 slides with 3 sections per slide. we cut 4 unstained between level 2 and 3 for ihc. Yes, we do have a lot of slides wasted also. However, since we have a histology program at the community college here, we usually give the school an slides that we would throw away (of course we are in accordance with all the rules and regulations with CAP and CLIA). Joe Nocito BS, HT (ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, Texas ----- Original Message ----- From: "Jennifer Englin" To: Sent: Monday, October 13, 2003 11:09 AM Subject: [Histonet] prostate needle biopsies We usually receive our needle biopsies in 2 contaners, right and left. I was wondering how many cassettes they are submitted in. For example 2 needles per cassette, five per cassette, etc... Also for those of you taking sections for possible IP's. We routinely cut five levels, 1,3,&5 for H&E and 2 & 4 for IP's. Do some of you cut seven??? Any help would be appreciated. Jennifer Englin, HT(ASCP),PA Rice Memorial Hospital Willmar, MN _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RSRICHMOND <@t> aol.com Mon Oct 13 21:22:37 2003 From: RSRICHMOND <@t> aol.com (RSRICHMOND@aol.com) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Ann Preece's estate Message-ID: <17f.21c7c62a.2cbcb7ed@aol.com> I got a most curious spam a week ago concerning the estate of Ann Preece. I suspect that a good many others on this list received it also. I reproduce it below. Those of us of a certain age remember Ann Preece, HT(ASCP), Scripps Mem. Hosp. in La Jolla CA, _A Manual for Histologic Technicians_, Little Brown & Co., 3rd edition 1972, ISBN 0-316-71765 (previous editions in 1965 and 1959). - This book was the book every histotech had, and studied for for the registry examination. Obviously dated, it's still worth having. Social Security Death Index gives a date of birth of 19 Nov 1923, date of death 26 Apr 2003 in San Diego. Wonder what books she had! Bob Richmond Samurai Pathologist Knoxville TN *************** >>HELLO: WE ARE ESTATE LIQUIDATORS IN SAN DIEGO. WE HAVE BEEN RETAINED TO LIQUIDATE THE ESTATE OF ANN PREECE AT 6095 AGEE. YOU MAY CONTACT US AT: 619-688-9414 OR 858-336-8510 << REGARDS; IGOR VON WURTTEMBURG E.F.WHALEN COMPANY ESTATE LIQUIDATOR WWW.EFWHALENCO.COM EFWhalenCo@aol.com October 7th, 2003 From hadi83 <@t> comcast.net Tue Oct 14 00:09:24 2003 From: hadi83 <@t> comcast.net (Hadi Yaziji) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] RE: negative IHC controls In-Reply-To: <004b01c391c8$4cb4f440$6400a8c0@bryaniwx13voft> Message-ID: <93A01C48-FE04-11D7-B67A-00039378E76A@comcast.net> Dear Bryan, Thanks for the instructive points about IHC QC. First, I do not represent the CAP (nor I want to), so I am speaking strictly from my own experience at PhenoPath and (prior to that) University of Washington and MD Anderson. You certainly have a point in terms of the "intended" use of controls to address a "full range of procedural variables" as you correctly put it. My concern with using QC in IHC is the end result. I can give you dozens of examples where the referring pathologists sent us cases in consultation, and we found errors in interpretation of studies, despite the presence of adequate positive and negative controls that accompanied their patient's slides. My point is a practical one: It is really the experience of the pathologist and technologist that is the rate-limiting factor that determines whether a given laboratory does an adequate job or not. This by no means nullifies the need for QC. It, instead, makes it more meaningful and cost-effective. What I just did is broaden the terms of QC, to include, in addition to the technical aspects, interpretation and validation aspects that, in my experience, are way more important (see discussion below on validation). Unfortunately, the CAP focuses (correctly) on the technical aspects, but pays very little attention to the interpretation aspects. The results is hundreds of labs passing the inspections every year but far fewer have comprehensive approach to technical, interpretation, validation and (I will not discuss this) proper selection of antibodies. I was very careful by saying that running multiple reagent control is IDEAL, but I didn't say it's practical, so I don't think I downplayed the complete reagent control issue. Regarding your comments on positive external controls: in real life, when the expected positive internal control fails, they usually fail once (for four main reasons). Repeat studies will yield adequate results except when the problem is tissue antigenicity due to fixation and processing issues. If that's the case, studies should be repeated using different (gentler or harsher) pretreatment. Putting positive external controls on the same slide will not make any difference in cases like this one. The expected positive internal control immunoreactivity is non-controversial. It is based on validation of the antibody in a given laboratory (not by the manufacturer) PRIOR to its approval as a diagnostic test. That's what I meant by money well spent. When we acquire any new antibody, the first thing we do is ignore the package insert, and test the antibody using at least 5 different pretreatments on tissues that are expected to be positive, and negative. Once we establish the proper pretreatment and antibody dilution, we run the antibody on a pilot study composed of X number of lesions. If the results are what we anticipate, then we validate the antibody and approve it for clinical use. That, in my opinion, is necessary and money well spent. But to run positive and negative controls on every slide is just not practical in most settings. I am delighted that some of my colleagues are interested in discussing this issue. I hope this note helped clarifying a couple of issues to the group. Hadi Yaziji, M.D. PhenoPath Laboratories On Monday, October 13, 2003, at 01:26 PM, Bryan Hewlett wrote: > Dear Hadi, > > I agree with much of the content of your posting. However, there seems > to be > a mixed message here. > You agree that; (a) running multiple reagent control sections covering > the > appropriate antibodies/pretreatments is ideal and that, (b) money > should be > well spent on QC (which you emphasize), yet you seem to downplay the > use of > complete reagent controls and to inject some confusion regarding the > necessary use of process controls. > > Process controls are intended to address the full range of procedural > variables and to ensure that the end result of the process is known to > lie > within legitimate or acceptable values and is valid. Process controls > are a > necessary tool utilized PRIOR to the subsequent interpretation of the > result. > Subsequent interpretation of the result depends largely upon the > validity > provided by these process controls. No one has suggested that a reagent > control on the patient slide is the only way to recognize a false > positive > signal. In fact, the experienced observer provides the final additional > level of process control. > However, that does not mean that all the necessary prior process > controls > can be avoided or omitted by that experienced observer. > We too, have seen 100's of IHC stains daily and the full use of process > controls has been an important factor in obtaining consistent results. > We have seen numerous occasions where reagent controls have provided > essential clues to the nature of an aberrant finding. > I would have to disagree with you that a single reagent control is > more than > sufficient. > > I do agree with your comments regarding the use of one positive > 'external' > tissue control per antibody/ per run but would point out that, if the > positive 'external' control works and the patient 'expected' positive > internal controls didn't, repeating the antibody test, using the same > process, should produce exactly the same result! > That would mean that either the 'expected' result is not a valid > expectation, or that the process controls are inadequate and do not > account > for all of the procedural variables. > > Regards, > > Bryan Hewlett > > > On Friday, October 10, 2003, at 10.06 PM, Hadi Yaziji wrote: > > Theoretically, I agree with you that (a) running multiple negative > control > sections to cover the different antibodies/pretreatments is ideal and > (b) > money should be well spent on QC (I can't emphasize that > enough). > However, I honestly don't remember when was the last time when I had a > false > positive staining on a patient's slide that the negative > control > was the only way that allowed me to recognize the false positive > signal on > the real slide. And we look at 100s of IHC stains every day. > ........ > > Experienced pathologists and technologists can still recognize in > > 99% of > the times a false positive signal from a real signal on the tissue > section > of a given antibody without even having to look at negative controls. > ... > > Individuals who perform and interpret IHC studies must have the > knowledge to > recognize the expected sub-cellular localization of every antibody on > every > type of tissue, including aberrant signals.... > > All you need is one positive control per antibody, but not one > control/per > antibody/per patient. In many cases positive internal controls are > present > on the same slide, so you can tell whether your antibody worked simply > by > evaluating the expected positive internal controls. In fact, if your > positive 'external' control worked and internal controls didn't, then > you > need to repeat your antibody test regardless of the positive external > control........ > > The bottom line: in real life, one negative control per case is more > than > sufficient. And to say the least, it's inaccurate to state this is poor > patient care and lousy quality control. > > > > > > > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From hadi83 <@t> comcast.net Tue Oct 14 00:42:49 2003 From: hadi83 <@t> comcast.net (Hadi Yaziji) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] RE: negative IHC controls In-Reply-To: Message-ID: <3EB23FAB-FE09-11D7-B67A-00039378E76A@comcast.net> Thanks for sharing your CV with us. You seem to be fixated on how slides should be filed. It's quite easy in our laboratory (and any other laboratory) to find the positive QC's, because they're filed chronologically (is there any other way to file slides?). First you find out the date the case in question was run, and you go to the QC file cabinet and pull the positive QC's that belong to the same day. Negative QC's (and again, one slide per tissue block) are filed with the case. As to your "evidence based good practice", please don't generalize. Two hospitals across the street from each other can represent two extremes of quality of care. Actually, I happened to have spent a month in a prestigious Histopathology Department in London nine years ago. The quality of IHC in their lab was superb, but it was miserable in the hospital right across the street from them. The same applies to the US and any other country. Finally, if my last email was unprofessional and insulting to you, please accept my apology. Please remember that it was your unnecessary use of the terms "dam fool" and "poor quality control by any definition of the term" in your previous email that initiated my response to you. Best wishes, Hadi Yaziji, M.D. PhenoPath Laboratories On Monday, October 13, 2003, at 03:00 PM, Nick Kirk wrote: > Hadi > ? > I am sorry but I must take issue with your very condescending and if I > may say so insulting tone. > ? > For your information, I have over 18 years of experience doing IHC and > my lab does a considerable number of slides per day. > I have set up the IHC service in two labs and have received much > praise for doing it. > I have also done extensive?lab based?trials work for both DAKO and > Novocastra here in the UK as well, so I think my credentials are > better than most. > I'm not at issue with the interpretation issue at all, that is indeed > a very skilled task that takes many years of study and experience to > become competent in. > What I am taking issue with is the issues around clinical audit and > best practice. > Now I don't know what clinical audit systems you have in your > establishment, but in ours it is regarded as poor clinical governance > i.e. bad practice by both myself, the Head Biomedical Scientist of the > department and all of my Pathologist colleagues not to have separate > controls per case. > This is especially the case when you are reviewing tumour antigenicity > on samples that may come out of the archive. > All our slides are stored in case order so to have positive controls > for each case actually makes it much easier the extract all the > relevant slides per case for later review. Having slides for the same > case filed in separate places introduces the potential for error, > especially when being initially filed. > I know this to be true because it has happened to me on several > occasions where slides filed in different places have been mis-filed > and become difficult to retrieve. > In the UK we use a system of evidence based good practice and evidence > suggests that is precisely what we are doing. > ? > I hope that now explains it to you. > ? > Nick Kirk BSc (Hons)?FIBMS (I've got letters too) > Head Biomedical Scientist > Histopathology > Hinchingbrooke Hospital > Huntingdon > Cambridgeshire > England > ? > > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Hadi > Yaziji > Sent: 11 October 2003 16:22 > To: histonet@pathology.swmed.edu > Subject: Re: [Histonet] negative IHC controls > > Dear Nick, > > Thanks for the email. You definitely misread my email. First, where in > my message did I advocate not to use negative and positive controls > per case? Of course it's poor patient's care not to do that. Is it > necessary to run + and - controls on every antibody? I don't think so > provided you have the necessary experience to interpret them. To > address your other point about reviewing cases, we keep our positive > and negative controls indefinitely. You can always pull the controls > that pertain to the run of the particular day of the case in question. > > You will be 'dam fool' (I am copying your words) to run IHC if you > have no experience. That's much more dangerous legally and ethically. > I've seen slides misinterpreted where the positive control is present > on the same slide. > > Bottom line: one + control per run is adequate (provided you have the > experience, which I'm not sure you have based on your reply). One - > control per case is more than sufficient. > > I hope this addresses your concerns. > > Hadi Yaziji, M.D. > PhenoPath Laboratories > > > On Saturday, October 11, 2003, at 12:16 AM, Nick Kirk wrote: > > A cautionary note > ? > I agree that false positives are rare but they do happen. We had a > couple a while ago when it turned out that the Teflon coat on the > probe of our DAKO Autostainer had perished and was carrying over > reagents from one slide to another and also contaminating our negative > control solution with primary antibody. > Also they are very useful in checking that there is no endogenous > biotin activity (more common than people think) and also that your > hydrogen peroxide solution hasn't gone off when quenching endogenous > peroxidase activity. > ? > As for the argument about only one positive control per antibody per > run rather than per patient - > Well the question I would pose is this. If in 5 years time you wish to > review the immuno for a particular case, how do you ensure that the > staining quality was adequate at the time if there aren't any positive > controls for that case? > On medico-legal grounds alone you should be doing positive controls > for each case, especially if those results end up with the patient > having a particularly aggressive treatment like chemotherapy or a > particularly invasive surgical procedure such as a colectomy or a > mastectomy. If someone wants to sue your organisation at a later date > for inappropriate treatment you need every bit of proof that your part > of the investigation was above reproach and controls is one thing that > will definitely be asked about. I certainly wouldn't like to stand up > in court and try and defend myself against that one! > ? > Incidentally, we use Surgipath's Control slides which allow you (most > of the time depending on the size of the test section) to have the > positive control and test section on the same slide, which saves a lot > of space on the immunostainer and neatly solves the problem of where > do you store the positive control if you use the single control per > run model and you have multiple cases with the same antibody. It also > acts as a check that the slide has received the correct primary > antibody and that someone hasn't loaded the immunostainer wrongly. > ? > At the end of the day I still go by the analogy someone else made here > earlier about airbags and seat belts in cars. Would you drive in a car > without them? They may never be needed in your entire driving life, > but you would be a dam fool not to have? them wouldn't you? > ? > And I?think we will have to agree to?disagree Hadi, with your last > statement, it is poor quality control by any definition of the term, > not to use both negative and positive controls for each case. > ? > Nick Kirk > Histopathology > Hinchingbrooke Hospital > Huntingdon > England > > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Hadi > Yaziji > Sent: 11 October 2003 03:07 > To: histonet@pathology.swmed.edu > Subject: Re: [Histonet] negative IHC controls > > Theoretically, I agree with you that (a) running multiple negative > control sections to cover the different antibodies/pretreatments is > ideal and (b) money should be well spent on QC (I can't emphasize that > enough). However, I honestly don't remember when was the last time > when I had a false positive staining on a patient's slide that the > negative control was the only way that allowed me to recognize the > false positive signal on the real slide. And we look at 100s of IHC > stains every day. > > Experienced pathologists and technologists can still recognize in > > 99% of the times a false positive signal from a real signal on the > tissue section of a given antibody without even having to look at > negative controls. I don't think it should be a CAP requirement at > all. In fact, I'd say the same thing on positive controls. All you > need is one positive control per antibody, but not one control/per > antibody/per patient. In many cases positive internal controls are > present on the same slide, so you can tell whether your antibody > worked simply by evaluating the expected positive internal controls. > In fact, if your positive 'external' control worked and internal > controls didn't, then you need to repeat your antibody test regardless > of the positive external control. External controls are different > tissues, fixed differently, processed differently and the tissue ages > differently (in terms of its antigenicity). > > Individuals who perform and interpret IHC studies must have the > knowledge to recognize the expected sub-cellular localization of every > antibody on every type of tissue, including aberrant signals. For > instance, you can easily dismiss a granular cytoplasmic TTF-1 signal > as not positive, because TTF-1 is a nuclear transcription factor and > we know it's expressed in the nuclei. However, hepatocellular > carcinomas can give you a consistent and reproducible granular and > cytoplasmic TTF-1 signal, and if you subsequently run confirmatory > markers (HepPar1, polyclonal CEA, CD10, etc.) they will be positive. > Such (granular cytoplasmic) signal should be recognized by the > interpreter (tech or pathologist) as a specific signal for tumors with > hepatoid phenotypes. This is just one of many examples.. > > The bottom line: in real life, one negative control per case is more > than sufficient. And to say the least, it's inaccurate to state this > is poor patient care and lousy quality control. > > I look forward to any constructive criticism. > > Hadi Yaziji, M.D. > PhenoPath Laboratories > > On Thursday, October 9, 2003, at 07:03 AM, Horn, Hazel V wrote: > > Not only should you be running a negative control for each patient > slide.?? That negative control should be treated just as your antibody > is.?? If the antibody is rabbit and antigen retrieved, so should your > control.?? If another antibody on the same patient is mouse and not > retrieved another negative control should be run with this same > protocol.?? In the United States, labs that are inspected by the CAP, > are required to run these controls.??? MONEY should never be > considered as a reason to stop doing a part of a procedure.?? It's > poor patient care and lousy quality control.?? IMHO. > ? > Hazel Horn, HT/HTL (ASCP) > Histology Supervisor > Arkansas Children's Hospital > > Phone - 501.364.4240 > Fax - 501.364.3912 > > > -----Original Message----- > From: vermast [mailto:vermast@rogers.com] > Sent: Wednesday, October 08, 2003 3:57 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] negative IHC controls > > ? > I would like to get a feel for how many out there are running negative > control slides for IHC.? > ? > In our lab we do just a handful of antibodies and initially I had been > running a negative control slide with each patient slide. ? After much > discussion with our pathologists, we decided to omit these negatives > (which were conistently negative)?and continue to just run a positive > control with each primary antibody for the run.? We use the Dako > autostainer and prediluted primaries.? The decision to stop running > negatives also coincided with Dako's decision to sell the negative > control sera separately from the primaries (they used to come packaged > together).? Perhaps I assumed that discontinuing to pair these > reagents together meant that few labs were using the negatives. > ? > Anyhow, after having reviewed the last QMPLS (Canada) survey committee > comments, I believe the committe would like a negative control run > with each patient tissue slide in order to evaluate background? (they > have used NCCLS guide pages as reference).? Incidentally we weren't a > part of the survey due to a technicality. > Any help or advice would be appreciated. > ? > L. Vermast > Stratford, Ont. > > > > > > > > > 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. Arkansas > Children's Hospital > > -------------- next part -------------- A non-text attachment was scrubbed... Name: not available Type: text/enriched Size: 15185 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031013/719b3500/attachment.bin From Ian.Bernard <@t> LACKLAND.AF.MIL Tue Oct 14 09:01:48 2003 From: Ian.Bernard <@t> LACKLAND.AF.MIL (Bernard Ian R SSgt 59 CRES/MSROP) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Help; looking for a stain receipe for TTC Message-ID: <588C513CC306D611A2910003479604F9065918B5@fsmpls17.whmc.af.mil> Hello folk or fellow histonetters(in particular in research), I'm in the process of replicating the histochemical staining method in a protocol article titled " LV-Powered Coronary Sinus Retroperfusion Reduces infarct Size in Acutely Ischemic Pigs." The made reagents for staining the heart tissue calls for a pre staining is a Phthalol-blue, which we will be substituting with Evans Blue. I need a concentration and receipe to make this up? The article does not provide this. The article states that the " ascending aorta was occluded and phthalo-blue(Evans Blue) was injected into the left ventricle. Finally the article states: " The left Ventricle was sliced, parallel to the atriopventricular groove, into several 5-10 mm slices and incubated for 30 mins at 37 deg C in TTC or Triphenyltetrazolium Chloride. There are 2 concentrations by Sigma, which one is to be used here, a 95% or 99.5 %? Need receipe to make this solution? What is Trisma HCL and Trisma Base? I need catalog #'s and source? The article goes on to state for results:That "the Mycardial tissue containing dehydrogenase enzymes is stained brick red by TTC, but areas of necrotic tissue depleted of these enzymes are not stained and appear as white or pale yellow." Thanks SSgt Bernie -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031014/e81120b1/attachment.htm From tpmorken <@t> labvision.com Tue Oct 14 10:35:22 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Help; looking for a stain receipe for TTC Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CC0A@usca0082k03.rallansci.apogent.com> SSgt Bernie If this (below) is the same article, I notice that they have an email given for contact. Maybe ask them the particulars. I can tell you that Trizma HCl and Trizma Base are from Sigma (look under Trizma in the chemical list). They supply it like this, in two parts, so you can mix the exact pH buffer you wish to use (between pH 7 and 9) . They give instructions with the chemicals. Another way to do it is to get Trizma pre-set crystals which will give you the exact pH you want with out any mixing of chemicals (between pH 7 and 9). Ann Thorac Surg 2000;69:84-89 (c) 2000 The Society of Thoracic Surgeons _____ LV-powered coronary sinus retroperfusion reduces infarct size in acutely ischemic pigs Jeffrey S. Martin, MDa, John G. Byrne, MDa, Olivier Y. Ghez, MDa, Umer Sayeed-Shah, MDa, Sergey D. Grachev, MDa, Rita G. Laurence, BSa, Lawrence H. Cohn, MDa a Division of Cardiac Surgery, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA Address reprint requests to Dr Byrne, Division of Cardiac Surgery, Brigham and Women's Hospital, 75 Francis St, Boston, MA 02115 e-mail: jgbyrne@bics.bwh.harvard.edu Background. We developed a prosthetic left ventricle (LV) to coronary sinus (CS) shunt (LVCSS) that is autoregulating and provides LV-powered retrograde perfusion of the coronary sinus. Methods. Each of 20 Yorkshire pigs underwent 1 hour of left anterior descending diagonal artery occlusion followed by 3 hours of reperfusion. The controls (n = 5) did not have shunt treatment. The LVCSS group (n = 9) underwent shunt treatment during the ischemic period. The LVCSS with partial coronary sinus occlusion (PCSO) group (LVCSS+PCSO, n = 6) underwent shunt treatment and PCSO during the ischemic period. Vital staining and planimetry techniques were used to determine the area at risk for infarction and the area of necrosis. Results. The area at risk was not significantly different among groups. The area of necrosis was decreased by 53% in the LVCSS group and by 73% in the LVCSS+PCSO group when compared to controls (p < 0.01 among all groups). Conclusions. The LVCSS reduces infarct size in pigs after acute coronary artery occlusion. The addition of PCSO to LVCSS further improves myocardial salvage. Tim Morken Lab Vision / NeoMarkers www.labvision.com -----Original Message----- From: Bernard Ian R SSgt 59 CRES/MSROP [mailto:Ian.Bernard@LACKLAND.AF.MIL] Sent: Tuesday, October 14, 2003 7:02 AM To: Histonet (E-mail) Subject: [Histonet] Help; looking for a stain receipe for TTC Importance: High Hello folk or fellow histonetters(in particular in research), I'm in the process of replicating the histochemical staining method in a protocol article titled " LV-Powered Coronary Sinus Retroperfusion Reduces infarct Size in Acutely Ischemic Pigs." The made reagents for staining the heart tissue calls for a pre staining is a Phthalol-blue, which we will be substituting with Evans Blue. I need a concentration and receipe to make this up? The article does not provide this. The article states that the " ascending aorta was occluded and phthalo-blue(Evans Blue) was injected into the left ventricle. Finally the article states: " The left Ventricle was sliced, parallel to the atriopventricular groove, into several 5-10 mm slices and incubated for 30 mins at 37 deg C in TTC or Triphenyltetrazolium Chloride. There are 2 concentrations by Sigma, which one is to be used here, a 95% or 99.5 %? Need receipe to make this solution? What is Trisma HCL and Trisma Base? I need catalog #'s and source? The article goes on to state for results:That "the Mycardial tissue containing dehydrogenase enzymes is stained brick red by TTC, but areas of necrotic tissue depleted of these enzymes are not stained and appear as white or pale yellow." Thanks SSgt Bernie -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031014/9b724a08/attachment.htm From pruegg <@t> msn.com Tue Oct 14 11:00:52 2003 From: pruegg <@t> msn.com (Patsy Ruegg) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] IHC QC's Message-ID: IHC QC will be a topic discussed at the IHC Resource Group committee meeting during the NSH S/C in Louisville. The committee meeting will be on Saturday at 5:30 PM. I invite all those attending the NSH S/C with an interest in IHC to attend this meeting. Best regards, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 HP-303-644-4538 HF-303-644-3377 hm email pruegg@msn.com wk email pruegg@colobio.com 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. ----Original Message Follows---- From: Patti Loykasek To: "Horn, Hazel V" ,histonet Subject: Re: [Histonet] IHC QC's Date: Mon, 13 Oct 2003 14:18:00 -0700 There was in fact, a post from Nick Kirk on running a positive control with each case. I do realize the CAP requirements and am familiar with the checklist. Patti Loykasek Phenopath Laboratories Seattle, WA > > > I don't think anyone said a POSTIVE control should be run with each slide. > We were talking about negative controls, I believe. > I just copied and pasted from the latest CAP survey in another email. > > > Hazel Horn, HT/HTL (ASCP) > Histology Supervisor > Arkansas Children's Hospital > > Phone - 501.364.4240 > Fax - 501.364.3912 >> >> -----Original Message----- >> From: Patti Loykasek [mailto:ploykasek@phenopath.com] >> Sent: Monday, October 13, 2003 10:52 AM >> To: histonet >> Subject: [Histonet] IHC QC's >> >> I'm glad that everyone is so concerned with both negative and positive IHC >> controls. There is certainly more than one side to this issue. I will say >> that I don't think a positive QC on every slide is absolutely necessary, for >> many reasons. If the QC is rare & precious, then it is a waste of resources. >> As is running a negative control for every possible technique permutation on >> small amounts of tumor. I would rather have slides with tumor left for >> additional studies than have wasted tumor sections on 4-6 negative controls. >> You can always evaluate non-specific staining on slides that have had an >> antibody applied & that are negative with that antibody. The CAP is specific >> that positive controls be used for each antibody - see CAP checklist >> ANP.22550. They do not specify for each slide. Since positive QC's should be >> kept filed for the same number of years as the patient slide & records, it >> should be possible to pull a QC slide from the IHC run for a particular >> slide. In the CAP comment on ANP.22550, the use of internal QC's is also >> mentioned. Although there are many ways of dealing with the issue of QC's, >> I'm sure we all want to do what is prudent, abide by the regulations, and >> increase the level of patient care. >> Just my 2 cents worth. >> >> Patti Loykasek >> Phenopath Laboratories >> Seattle, WA > > 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. Arkansas Children's Hospital > _________________________________________________________________ Enjoy MSN 8 patented spam control and more with MSN 8 Dial-up Internet Service. Try it FREE for one month! http://join.msn.com/?page=dept/dialup From HornHV <@t> archildrens.org Tue Oct 14 14:03:03 2003 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Ann Preece's estate Message-ID: I wonder too. I know when I took my HT registry 30 years ago this year, her book and the AFIP staining manual were all we had. I still have my copy of Ann Preece. Now it's even more precious. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: RSRICHMOND@aol.com [mailto:RSRICHMOND@aol.com] Sent: Monday, October 13, 2003 9:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Ann Preece's estate I got a most curious spam a week ago concerning the estate of Ann Preece. I suspect that a good many others on this list received it also. I reproduce it below. Those of us of a certain age remember Ann Preece, HT(ASCP), Scripps Mem. Hosp. in La Jolla CA, _A Manual for Histologic Technicians_, Little Brown & Co., 3rd edition 1972, ISBN 0-316-71765 (previous editions in 1965 and 1959). - This book was the book every histotech had, and studied for for the registry examination. Obviously dated, it's still worth having. Social Security Death Index gives a date of birth of 19 Nov 1923, date of death 26 Apr 2003 in San Diego. Wonder what books she had! Bob Richmond Samurai Pathologist Knoxville TN *************** >>HELLO: WE ARE ESTATE LIQUIDATORS IN SAN DIEGO. WE HAVE BEEN RETAINED >>TO LIQUIDATE THE ESTATE OF ANN PREECE AT 6095 AGEE. YOU MAY CONTACT US AT: 619-688-9414 OR 858-336-8510 << REGARDS; IGOR VON WURTTEMBURG E.F.WHALEN COMPANY ESTATE LIQUIDATOR WWW.EFWHALENCO.COM EFWhalenCo@aol.com October 7th, 2003 _______________________________________________ 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. Arkansas Children's Hospital From HornHV <@t> archildrens.org Tue Oct 14 14:07:37 2003 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] IHC QC's Message-ID: For those of us who will not be attending NSH will you send us a copy of the meetings minutes? Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: Patsy Ruegg [mailto:pruegg@msn.com] Sent: Tuesday, October 14, 2003 11:01 AM To: ploykasek@phenopath.com; HornHV@archildrens.org; histonet@pathology.swmed.edu Subject: Re: [Histonet] IHC QC's IHC QC will be a topic discussed at the IHC Resource Group committee meeting during the NSH S/C in Louisville. The committee meeting will be on Saturday at 5:30 PM. I invite all those attending the NSH S/C with an interest in IHC to attend this meeting. Best regards, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 HP-303-644-4538 HF-303-644-3377 hm email pruegg@msn.com wk email pruegg@colobio.com 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. ----Original Message Follows---- From: Patti Loykasek To: "Horn, Hazel V" ,histonet Subject: Re: [Histonet] IHC QC's Date: Mon, 13 Oct 2003 14:18:00 -0700 There was in fact, a post from Nick Kirk on running a positive control with each case. I do realize the CAP requirements and am familiar with the checklist. Patti Loykasek Phenopath Laboratories Seattle, WA > > > I don't think anyone said a POSTIVE control should be run with each slide. > We were talking about negative controls, I believe. > I just copied and pasted from the latest CAP survey in another email. > > > Hazel Horn, HT/HTL (ASCP) > Histology Supervisor > Arkansas Children's Hospital > > Phone - 501.364.4240 > Fax - 501.364.3912 >> >> -----Original Message----- >> From: Patti Loykasek [mailto:ploykasek@phenopath.com] >> Sent: Monday, October 13, 2003 10:52 AM >> To: histonet >> Subject: [Histonet] IHC QC's >> >> I'm glad that everyone is so concerned with both negative and positive IHC >> controls. There is certainly more than one side to this issue. I will say >> that I don't think a positive QC on every slide is absolutely necessary, for >> many reasons. If the QC is rare & precious, then it is a waste of resources. >> As is running a negative control for every possible technique permutation on >> small amounts of tumor. I would rather have slides with tumor left for >> additional studies than have wasted tumor sections on 4-6 negative controls. >> You can always evaluate non-specific staining on slides that have had an >> antibody applied & that are negative with that antibody. The CAP is specific >> that positive controls be used for each antibody - see CAP checklist >> ANP.22550. They do not specify for each slide. Since positive QC's should be >> kept filed for the same number of years as the patient slide & records, it >> should be possible to pull a QC slide from the IHC run for a particular >> slide. In the CAP comment on ANP.22550, the use of internal QC's is also >> mentioned. Although there are many ways of dealing with the issue of QC's, >> I'm sure we all want to do what is prudent, abide by the regulations, and >> increase the level of patient care. >> Just my 2 cents worth. >> >> Patti Loykasek >> Phenopath Laboratories >> Seattle, WA > > 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. Arkansas Children's Hospital > _________________________________________________________________ Enjoy MSN 8 patented spam control and more with MSN 8 Dial-up Internet Service. Try it FREE for one month! http://join.msn.com/?page=dept/dialup 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. Arkansas Children's Hospital From CWilsonCPI <@t> ameripath.com Tue Oct 14 14:49:56 2003 From: CWilsonCPI <@t> ameripath.com (CWilsonCPI@ameripath.com) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Temporary full-time histotech position and per diem cytoprep tech position in northeast ohio Message-ID: We have a temporary full-time position for a histotechnician lasting for approximately four months with the possibility of turning into a permanent position. We are located outside of Cleveland, OH. At this time I'm NOT looking to fill this position through an agency. So if you are affiliated with an agency do NOT contact me at this time. Anyone else interested please fax a resume to my attention at the number below. We also have a per diem position for a cytology prep person interested in working a few hours a week and to provide coverage for approximately 4-6 weeks full- time in December. Fax resumes for this position also to the number below. Thanks, Carol Wilson Lab Supervisor AmeriPath Cleveland Fax 440-703-2155 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 KHays <@t> mbhs.org Tue Oct 14 15:00:28 2003 From: KHays <@t> mbhs.org (KHays@mbhs.org) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] hazards of isopentane 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 one of my pathologist wants to start snap freezing our frozens with dry ice and isopentane. what are the hazards in working with this and what equipment is needed? thanks in advance for any information. The information contained in this email is confidential and is intended for the recipient only. Do not forward this email without permission from its originator. From sschmoll <@t> umich.edu Tue Oct 14 15:53:13 2003 From: sschmoll <@t> umich.edu (Stephanie Schmoll) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Mouse Embryo Processing Message-ID: <6D35C6F8-FE88-11D7-8646-00039375B984@umich.edu> I am fairly new to tissue processing with paraffin, have always done cryo-embedding in the past. I am having trouble processing late stage mouse embryos, day 17 and 18. The paraffin doesn't seem to be infiltrating. Any suggestions would be appreciated. My current protocol is as follows: 24hrs 10% NBF, 70% EtOH 30 min, 80% 30 min, 95% 30 min x2, 100% 30 min x2, Xylene 30 min x2, paraffin 30 min x3. Stephanie Schmoll, Morphology Core Manager Center for Organogenesis University of Michigan Medical School 5795C Medical Science Bldg. II Phone: (734) 647-9021 Fax: (734) 647-9559 From David.Muskett <@t> btopenworld.com Tue Oct 14 17:08:28 2003 From: David.Muskett <@t> btopenworld.com (David Muskett) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Rhabdomyosarcomas Message-ID: Skipped content of type multipart/alternative-------------- next part -------------- A non-text attachment was scrubbed... Name: not available Type: image/gif Size: 530 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031014/5ed4b471/attachment.gif From mari.ann.mailhiot <@t> leica-microsystems.com Tue Oct 14 17:38:34 2003 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Mouse Embryo Processing Message-ID: Stephanie It may be the type of paraffin you are using. Surgipath makes a great infiltration medium for the processor. It is called infiltration medium. The embedding medium would be EM 400. I would also add some time to the paraffin - 45 min. x3. Something else you may want to think about is using a xylene substitute instead of xylene. Hope this helps. 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 Stephanie Schmoll To: histonet@lists.utsouthwestern.edu Sent by: cc: histonet-admin@lists.utsouth Subject: [Histonet] Mouse Embryo Processing western.edu 10/14/2003 03:53 PM I am fairly new to tissue processing with paraffin, have always done cryo-embedding in the past. I am having trouble processing late stage mouse embryos, day 17 and 18. The paraffin doesn't seem to be infiltrating. Any suggestions would be appreciated. My current protocol is as follows: 24hrs 10% NBF, 70% EtOH 30 min, 80% 30 min, 95% 30 min x2, 100% 30 min x2, Xylene 30 min x2, paraffin 30 min x3. Stephanie Schmoll, Morphology Core Manager Center for Organogenesis University of Michigan Medical School 5795C Medical Science Bldg. II Phone: (734) 647-9021 Fax: (734) 647-9559 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________ This email has been scanned for all viruses by the MessageLabs Email Security System. For more information on a proactive email security service working around the clock, around the globe, visit http://www.messagelabs.com ________________________________________________________________________ From pruegg <@t> msn.com Tue Oct 14 17:44:31 2003 From: pruegg <@t> msn.com (Patsy Ruegg) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] IHC QC's Message-ID: Hazel, The minutes for the IHCRG committee meeting will be posted on the IHCRG web site (www.ihcrg.org)and on our list serve ihcrg@yahoogroups.com for members of the group. I think you are a member but if not and if you are not receiving the messages from our list serve you can apply for membership with the group at www.ihcrg.org Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 HP-303-644-4538 HF-303-644-3377 hm email pruegg@msn.com wk email pruegg@colobio.com 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. ----Original Message Follows---- From: "Horn, Hazel V" To: 'Patsy Ruegg' , ploykasek@phenopath.com, "Horn, Hazel V" , histonet@pathology.swmed.edu Subject: RE: [Histonet] IHC QC's Date: Tue, 14 Oct 2003 14:07:37 -0500 For those of us who will not be attending NSH will you send us a copy of the meetings minutes? Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: Patsy Ruegg [mailto:pruegg@msn.com] Sent: Tuesday, October 14, 2003 11:01 AM To: ploykasek@phenopath.com; HornHV@archildrens.org; histonet@pathology.swmed.edu Subject: Re: [Histonet] IHC QC's IHC QC will be a topic discussed at the IHC Resource Group committee meeting during the NSH S/C in Louisville. The committee meeting will be on Saturday at 5:30 PM. I invite all those attending the NSH S/C with an interest in IHC to attend this meeting. Best regards, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 HP-303-644-4538 HF-303-644-3377 hm email pruegg@msn.com wk email pruegg@colobio.com 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. ----Original Message Follows---- From: Patti Loykasek To: "Horn, Hazel V" ,histonet Subject: Re: [Histonet] IHC QC's Date: Mon, 13 Oct 2003 14:18:00 -0700 There was in fact, a post from Nick Kirk on running a positive control with each case. I do realize the CAP requirements and am familiar with the checklist. Patti Loykasek Phenopath Laboratories Seattle, WA > > > I don't think anyone said a POSTIVE control should be run with each slide. > We were talking about negative controls, I believe. > I just copied and pasted from the latest CAP survey in another email. > > > Hazel Horn, HT/HTL (ASCP) > Histology Supervisor > Arkansas Children's Hospital > > Phone - 501.364.4240 > Fax - 501.364.3912 >> >> -----Original Message----- >> From: Patti Loykasek [mailto:ploykasek@phenopath.com] >> Sent: Monday, October 13, 2003 10:52 AM >> To: histonet >> Subject: [Histonet] IHC QC's >> >> I'm glad that everyone is so concerned with both negative and positive IHC >> controls. There is certainly more than one side to this issue. I will say >> that I don't think a positive QC on every slide is absolutely necessary, for >> many reasons. If the QC is rare & precious, then it is a waste of resources. >> As is running a negative control for every possible technique permutation on >> small amounts of tumor. I would rather have slides with tumor left for >> additional studies than have wasted tumor sections on 4-6 negative controls. >> You can always evaluate non-specific staining on slides that have had an >> antibody applied & that are negative with that antibody. The CAP is specific >> that positive controls be used for each antibody - see CAP checklist >> ANP.22550. They do not specify for each slide. Since positive QC's should be >> kept filed for the same number of years as the patient slide & records, it >> should be possible to pull a QC slide from the IHC run for a particular >> slide. In the CAP comment on ANP.22550, the use of internal QC's is also >> mentioned. Although there are many ways of dealing with the issue of QC's, >> I'm sure we all want to do what is prudent, abide by the regulations, and >> increase the level of patient care. >> Just my 2 cents worth. >> >> Patti Loykasek >> Phenopath Laboratories >> Seattle, WA > > 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. Arkansas Children's Hospital > _________________________________________________________________ Enjoy MSN 8 patented spam control and more with MSN 8 Dial-up Internet Service. Try it FREE for one month! http://join.msn.com/?page=dept/dialup 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. Arkansas Children's Hospital _________________________________________________________________ Never get a busy signal because you are always connected with high-speed Internet access. Click here to comparison-shop providers. https://broadband.msn.com From BoozerKA <@t> pa1.ah.org Tue Oct 14 18:50:30 2003 From: BoozerKA <@t> pa1.ah.org (Kathleen Boozer) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Full time 6-2:30 openning Message-ID: Adventist Medical Center, Portland, OR From AnthonyH <@t> chw.edu.au Tue Oct 14 21:31:08 2003 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Demonstration of Methylmalonic acid crystals Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E05B@simba.kids> Hi all, I have had a request for the histological demonstration of Methylmalonic acid crystals in a renal biopsy. I have been unable to find a procedure. Would anyone have any insights or references they could share? Yours in Histo, 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 georgecole <@t> ev1.net Wed Oct 15 02:01:11 2003 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Thumbnail list of muscle biopsy errors Message-ID: <000001c392ea$1eb75600$094dbad0@hppav> 1. Even when a substantial cubic cm piece of muscle was sent, tiny 1 and 2 mm bits of tissue were sectioned. 2. Gum Tragacanth was used to position the tissue Tragacanth helps get good sections., but it is a bit messy and it is itself a form of sugar which tends to confound testing for abnormal glycogen. 3. To hold the tissue for freezing odd heavy metal plugs were used with gum tragacanth and pip of muscle on top of it to insert into the Isopentane and liquid nitrogen. The mass of metal commands the edge of the cold effect, which tends to add to the threat of ice crystals. 4. Gum tragacanth surrounding bits of tissue are placed on cork to freeze. Cork is a very good insulator and adds a bit to the chance of ice crystals on the underside of the tissue. 5. Isopentane is cooled in the liquid nitrogen until it gets a little thick. 6. The tissues are sectioned in their native shape which will almost always be odd, with planes and peeks of tissue. Sections are hardly at their best as they almost always twist and curl onto the knife due to the erratic angles presented to the knife. You can see the twisting wayward sections coming onto the knife in some of the muscle text books. 7. Sections---for some strange reason, perhaps due to the suggestion of a tricky leprechaun, who is still laughing at us for taking him up on it----are picked up on cover slips. 8. Amounts of reagents are kept to the minimum----barely covering the sections on the slide. 9. The usual buffer for ATP-ase mixtures continues to be the sodium pentobarbital buffer in the literature. This takes a controlled substance permit to procure and it's not worth the trouble. It's not the best buffer for ATP-ase. Suggestions on better ways will follow. This is already a lumpy message. . -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031015/84b4e58d/attachment.htm From Inga.Hansson <@t> neuro.uu.se Wed Oct 15 03:59:35 2003 From: Inga.Hansson <@t> neuro.uu.se (Inga Hansson) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] fixation of cryosections Message-ID: Hi everyone! Can anyone suggest fixation for cryosections other than PFA or methanol/aceton for IHC? Regards Inga -- Inga Hansson Dept. neuroscience, div. neurobiology PO Box 587, BMC Husargatan 3 S-751 23 Uppsala SWEDEN Phone: +46(18) 471 4384 Fax: +46(18)559017 From docmichel <@t> netbulmail.com Wed Oct 15 07:17:18 2003 From: docmichel <@t> netbulmail.com (izzet oguz) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] AEC Message-ID: <3303.193.255.128.130.1066220238.webmail@mail1.netbulmail.com> Hi there, we all used DAB for IHC before..but now we have an AEC kit and we suppose that dehidration isn't used for AEC but what about xylol?..do we put the slides in xylol after counterstain or not? Is there any difference apart from that..Will you please share your experiences?.. Thanks ?zzet From tissuearray <@t> hotmail.com Wed Oct 15 07:32:11 2003 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Tissue Microarrays and the Block Warmer Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031015/49b61c4a/attachment.htm From MMcCOY <@t> lakelandregional.org Wed Oct 15 07:33:13 2003 From: MMcCOY <@t> lakelandregional.org (Mary McCoy) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Morgue regulating agency Message-ID: Does anyone on the histonet have responsibilities for morgues? If so do you know the regulating agency that requires negative air pressure for morgues and / or the classification of morgues as to which types of autopsies (levels of infection) can be performed in each. Thanks in advance, Mary Mary McCoy Coordinator of Pathology Services Lakeland Regional Health System St. Joseph MI 49098 (269) 982-4891 -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:McCoy, Mary TEL;WORK:982-4891 ORG:;Lab TEL;PREF;FAX:98-8258 EMAIL;WORK;PREF;NGW:MMcCOY@lakelandregional.org N:McCoy;Mary TITLE:Coordinator Pathology END:VCARD BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:McCoy, Mary TEL;WORK:982-4891 ORG:;Lab TEL;PREF;FAX:98-8258 EMAIL;WORK;PREF;NGW:MMcCOY@lakelandregional.org N:McCoy;Mary TITLE:Coordinator Pathology END:VCARD From DDDeltour <@t> sig.med.navy.mil Wed Oct 15 08:03:20 2003 From: DDDeltour <@t> sig.med.navy.mil (Deltour, Douglas D.(HM2)) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Morgue regulating agency Message-ID: I believe that it is OSHA. We always go through our Industrial Hygiene dept. for neg pressure monitoring. Here is a website that might help. http://www.osha.gov/SLTC/etools/hospital/lab/lab.html#Morgue HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy LPO Histology Histology Technician DSN 624-4669 FAX 624-4680 COMM (From US) 011-39-095-56-4669 -----Original Message----- From: Mary McCoy [mailto:MMcCOY@lakelandregional.org] Sent: Wednesday, October 15, 2003 2:33 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Morgue regulating agency Does anyone on the histonet have responsibilities for morgues? If so do you know the regulating agency that requires negative air pressure for morgues and / or the classification of morgues as to which types of autopsies (levels of infection) can be performed in each. Thanks in advance, Mary Mary McCoy Coordinator of Pathology Services Lakeland Regional Health System St. Joseph MI 49098 (269) 982-4891 From sschmoll <@t> umich.edu Wed Oct 15 08:07:20 2003 From: sschmoll <@t> umich.edu (Stephanie Schmoll) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Mouse Embryo Processing In-Reply-To: <003501c3929a$b520f240$4800a8c0@PMARCUM2K> Message-ID: <827A018E-FF10-11D7-B73D-00039375B984@umich.edu> Sorry I'm new at the histonet! I am using a Leica ASP 300 tissue processor. All of the reagents run at room temp and paraffin is at 57C suitable for the Tissue Prep2 paraffin I am using. I am embedding whole embryos developmental days 7 through 18 and neonate. I am only having problems with the late stages. On Tuesday, October 14, 2003, at 05:32 PM, Pamela Marcum wrote: > Stephanie, > > Are these whole embryos? Do they have all of the skin in place? Can > you > give a size estimate? Are you using a tissue processor and if so > where are > you using heat (other than paraffin) and vacuum? What kind of tissue > processor? Please give temperatures also. Thanks, Pam Marcum > >> -----Original Message----- >> From: histonet-admin@lists.utsouthwestern.edu >> [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Stephanie >> Schmoll >> Sent: Tuesday, October 14, 2003 4:53 PM >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] Mouse Embryo Processing >> >> >> I am fairly new to tissue processing with paraffin, have always done >> cryo-embedding in the past. I am having trouble processing late stage >> mouse embryos, day 17 and 18. The paraffin doesn't seem to be >> infiltrating. Any suggestions would be appreciated. >> >> My current protocol is as follows: 24hrs 10% NBF, 70% EtOH 30 min, >> 80% >> 30 min, 95% 30 min x2, 100% 30 min x2, Xylene 30 min x2, paraffin 30 >> min x3. >> >> >> Stephanie Schmoll, Morphology Core Manager >> Center for Organogenesis >> University of Michigan Medical School >> 5795C Medical Science Bldg. II >> Phone: (734) 647-9021 >> Fax: (734) 647-9559 >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > Stephanie Schmoll, Morphology Core Manager Center for Organogenesis University of Michigan Medical School 5795C Medical Science Bldg. II Phone: (734) 647-9021 Fax: (734) 647-9559 From mcauliff <@t> umdnj.edu Wed Oct 15 08:45:13 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] fixation of cryosections In-Reply-To: References: Message-ID: <3F8D4F69.9040102@umdnj.edu> Hi Inga: You could use any fixative you want to. How about neutral buffered formalin or an alcoholic formalin? Why do you want to avoid the ones you mentioned? Are you getting poor results with your IHC? Geoff Inga Hansson wrote: > Hi everyone! > > Can anyone suggest fixation for cryosections other than PFA or > methanol/aceton for IHC? > > Regards > > Inga -- -- ********************************************** 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 MGondo <@t> popmail.opt.uh.edu Wed Oct 15 09:14:48 2003 From: MGondo <@t> popmail.opt.uh.edu (Margaret Gondo) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Flatbed Scanners for EM negatives Message-ID: <200310150914.AA104464676@popmail.opt.uh.edu> Hi all - I was just wondering if anyone out in Histoland has any recommendations on flatbed scanners that will do a decent job of scanning an EM negative. Our lab uses mostly Macintosh computers for imaging. I will admit that I know very little about scanners. I just visited a site that had film scanners that cost in the thousands along with flatbed scanners with similar DPI for a couple hundred. Any information would be greatly appreciated. Margaret From cfavara <@t> niaid.nih.gov Wed Oct 15 09:05:24 2003 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Help; looking for a stain recipe for TTC Message-ID: You might want to call the author or contact person on the paper. Protocols are usually the first on the cutting floor. Most people are delighted to have someone pursue their research! Good luck- c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: Bernard Ian R SSgt 59 CRES/MSROP [mailto:Ian.Bernard@LACKLAND.AF.MIL] Sent: Tuesday, October 14, 2003 8:02 AM To: Histonet (E-mail) Subject: [Histonet] Help; looking for a stain receipe for TTC Importance: High Hello folk or fellow histonetters(in particular in research), I'm in the process of replicating the histochemical staining method in a protocol article titled " LV-Powered Coronary Sinus Retroperfusion Reduces infarct Size in Acutely Ischemic Pigs." The made reagents for staining the heart tissue calls for a pre staining is a Phthalol-blue, which we will be substituting with Evans Blue. I need a concentration and receipe to make this up? The article does not provide this. The article states that the " ascending aorta was occluded and phthalo-blue(Evans Blue) was injected into the left ventricle. Finally the article states: " The left Ventricle was sliced, parallel to the atriopventricular groove, into several 5-10 mm slices and incubated for 30 mins at 37 deg C in TTC or Triphenyltetrazolium Chloride. There are 2 concentrations by Sigma, which one is to be used here, a 95% or 99.5 %? Need receipe to make this solution? What is Trisma HCL and Trisma Base? I need catalog #'s and source? The article goes on to state for results:That "the Mycardial tissue containing dehydrogenase enzymes is stained brick red by TTC, but areas of necrotic tissue depleted of these enzymes are not stained and appear as white or pale yellow." Thanks SSgt Bernie -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031015/91b92444/attachment.htm From metsolutions <@t> yahoo.com Wed Oct 15 10:21:21 2003 From: metsolutions <@t> yahoo.com (Martin - MetLab Solutions) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Histology equipment for sale Message-ID: <20031015152121.72793.qmail@web21402.mail.yahoo.com> Hello Everyone, We have some surplus equipment (very good condition) that we would like to sell off: a - Olympus BX-40 microscope b - Olympus BX-60 microscope c - Olympus SZX9 stereoscope d - 41 stainless steel staining racks (60 slide cap.) e - 8 staining troughs f - 2 microscope slide cabinets g - 4 packs Scienceware microscope slide view packs h - 5 microscope slide cases i - 2 acrylic dessicator cabinets j - 2 Pyrex vacuum dessicators k - Nuaire biological hood l - MonoAir ductless hood m - Hermle benchtop centrifuge n - analytical balances o - MasterFlex pumps Please send me an e-mail and we could send you some photos and prices. Regards Martin __________________________________ Do you Yahoo!? The New Yahoo! Shopping - with improved product search http://shopping.yahoo.com From HornHV <@t> archildrens.org Wed Oct 15 11:09:58 2003 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Ann Preece's estate Message-ID: I found this on a google search..... PREECE, ANN Ann Preece, born November 12, 1923, in Boston, Massachusetts, died in La Jolla, April 26, 2003. A resident of La Jolla and San Diego for many years, she worked at the pathology lab at Scripps Memorial Hospital and authored A Manual for Histological Technicians, published in three editions by Little, Brown & Co. Ann enjoyed many facets of life and excelled as a scientist, a seamstress, a belly dancer, a culinary specialist, an intellectual, and above all a devoted mother to her two sons, Richard and David Preece, both of whom predeceased her. She was tirelessly inquisitive, devoutly spiritual, and full of fun and wry humor. She is survived by her long-time companion Angelo Anastasion of San Diego, and her cousin Eleanor Joaquin of Boston. She will be greatly missed by loving friends and family. No services are planned locally. Memorial contributions may be made to charities of the donor's choice. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: RSRICHMOND@aol.com [mailto:RSRICHMOND@aol.com] Sent: Tuesday, October 14, 2003 9:05 PM To: HornHV@archildrens.org Subject: Re: [Histonet] Ann Preece's estate Hi Hazel Horn! >>I wonder too. I know when I took my HT registry 30 years ago this year, her book and the AFIP staining manual were all we had. I still have my copy of Ann Preece. Now it's even more precious.<< Wonder if an obituary will appear anywhere - perhaps the Journal of Histotechnology. I don't recall ever meeting anyone who knew her. Bob Richmond 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. Arkansas Children's Hospital From pruegg <@t> msn.com Wed Oct 15 10:27:43 2003 From: pruegg <@t> msn.com (Patsy Ruegg) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] IHCRG member applications Message-ID: I have just received a bunch of applications for membership to the IHC Resource Group. I just wanted to let you all know that I did receive them but they will not be processed until after the NSH S/C because the NSH office staff is not available . Membership in the group is not required to attend our committee meeting in Louisville. Anyone attending the meeting is welcome to the committee meeting on Sat. Oct. 18th at 5:30 PM. Best regards, Patsy Ruegg, IHCRG Chair Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 HP-303-644-4538 HF-303-644-3377 hm email pruegg@msn.com wk email pruegg@colobio.com 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. _________________________________________________________________ Get 10MB of e-mail storage! Sign up for Hotmail Extra Storage. http://join.msn.com/?PAGE=features/es From HornHV <@t> archildrens.org Wed Oct 15 11:09:58 2003 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Ann Preece's estate Message-ID: I found this on a google search..... PREECE, ANN Ann Preece, born November 12, 1923, in Boston, Massachusetts, died in La Jolla, April 26, 2003. A resident of La Jolla and San Diego for many years, she worked at the pathology lab at Scripps Memorial Hospital and authored A Manual for Histological Technicians, published in three editions by Little, Brown & Co. Ann enjoyed many facets of life and excelled as a scientist, a seamstress, a belly dancer, a culinary specialist, an intellectual, and above all a devoted mother to her two sons, Richard and David Preece, both of whom predeceased her. She was tirelessly inquisitive, devoutly spiritual, and full of fun and wry humor. She is survived by her long-time companion Angelo Anastasion of San Diego, and her cousin Eleanor Joaquin of Boston. She will be greatly missed by loving friends and family. No services are planned locally. Memorial contributions may be made to charities of the donor's choice. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: RSRICHMOND@aol.com [mailto:RSRICHMOND@aol.com] Sent: Tuesday, October 14, 2003 9:05 PM To: HornHV@archildrens.org Subject: Re: [Histonet] Ann Preece's estate Hi Hazel Horn! >>I wonder too. I know when I took my HT registry 30 years ago this year, her book and the AFIP staining manual were all we had. I still have my copy of Ann Preece. Now it's even more precious.<< Wonder if an obituary will appear anywhere - perhaps the Journal of Histotechnology. I don't recall ever meeting anyone who knew her. Bob Richmond 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. Arkansas Children's Hospital From tvedilago <@t> system1.net Wed Oct 15 12:13:30 2003 From: tvedilago <@t> system1.net (Tommy Vedilago) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Jobs in Histology Message-ID: Hello Histonetters, I just wanted to let everyone know some of the positions we are working on. Histology Manager/Supervisor in Tampa, Fl, Camden, NJ, Northern New Jersey, Arizona, Philadelphia, and New York. We also have tech positions in Nashville, Philadelphia, Michigan, South Florida, northern New Jersey, Arizona, Georgia, and Dallas, TX. I will be wandering around the NSH meeting exhibit area also and would love to meet as many of you as I can. Please feel free to look me up or you can call me and I will make some time to sit down with you while there. I can be reached at 866-SYSTEM1 (866-797-8361). Sincerely, Tommy Vedilago System 1 Search (864) 627-0012 (864) 627-0013 Fax From kbowden <@t> ucsd.edu Wed Oct 15 12:25:18 2003 From: kbowden <@t> ucsd.edu (K. Bowden) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Flatbed Scanners for EM negatives References: <200310150914.AA104464676@popmail.opt.uh.edu> Message-ID: <3F8D82FE.10401@ucsd.edu> We use Macintosh computers and do most all of our scanning on an AGFA Arcus II. I'm not sure if they sell that model any longer but I'm sure the latest model would have at lest the same quality if not better. We scan x-rays, 35 mm slides, we have even placed MMA embedded sections directly in the scanner and got excellent results. -- Karen Bowden Staff Research Associate II University of CA, San Diego Department of Orthopedics 9500 Gilman Dr. 0630 La Jolla, CA 92093-0630 858-277-4970 CONFIDENTIALITY NOTICE: 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. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVED THIS E-MAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER. Margaret Gondo wrote: > Hi all - > > I was just wondering if anyone out in Histoland has any > recommendations on flatbed scanners that will do a decent job of > scanning an EM negative. Our lab uses mostly Macintosh computers > for imaging. I will admit that I know very little about scanners. > I just visited a site that had film scanners that cost in the > thousands along with flatbed scanners with similar DPI for a couple > hundred. > > Any information would be greatly appreciated. > > Margaret > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From tissuearray <@t> hotmail.com Wed Oct 15 12:22:49 2003 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:22:03 2005 Subject: Fwd: Re: [Histonet] Tissue Microarrays and the Block Warmer Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031015/692e4e09/attachment.htm From JBurrill <@t> criver.com Wed Oct 15 12:27:55 2003 From: JBurrill <@t> criver.com (JBurrill@criver.com) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Mouse Embryo Processing Message-ID: Hi Stephanie, It depends on what you are looking for. If you are interested in multiple organs the most consistent way of processing is trimming the embryo into three mid-siggital sections for gestational days 15-18. Two on either side of the umbilicus and the umbilicus section. I have processed a lot of embryos and found this is the most consistent reproducible processing method. I would also agree that you need more time in the xylene, we use 3 stations for 45 min each. Jason D. Burrill, B.A., HT(ASCP) Senior Supervisor, Histology Charles River Laboratories 251 Ballardvale Street Wilmington, MA 01887 ph: 978-658-6000 ext 1652 fax: 978-988-8793 E-mail: jburrill@criver.com -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031015/246a8c32/attachment.htm From algranth <@t> u.arizona.edu Wed Oct 15 12:50:57 2003 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] fixative for insects Message-ID: <4.3.2.7.2.20031015104722.00cc80f0@algranth.inbox.email.arizona.edu> I'm going to be processing whole mosquitoes and I was wondering what would be the best fixative to use...and processing schedule if anybody has one. Thanks. Andi ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From georgecole <@t> ev1.net Wed Oct 15 12:54:41 2003 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Suggested corrections for list of errors Message-ID: <000001c39345$69cec190$074dbad0@hppav> 1. ALL of the tissues sent will be frozen and sectioned. There is no way to look at 100% of the tissues---but we can represent every portion of the tissues. With normally sized biopsies, pieces to be cut may be just slightly narrower than the cover glass or a group of several narrow pieces just narrower than the cover glass can be frozen side by side. Pieces are best processed if they are no more than half a cm thick and long. Huge biopsies a cubic inch and larger occasionally arrive. Pieces a cm long and almost a cm thick have been frozen all right, with occasional minor ice crystals in the center. As much of each segment of the tissue should be represented as well as possible in the sections. 2. OCT is the mounting medium of choice. It is compatible with all of the histochemistry. Only a thin coating of OCT is put in the bottom of the mold to hold the tissue in the mold during freezing. 3. Those thin walled plastic molds work just fine---they have a name, but I never knew it--- They come in at least two different sizes. About a ? inch square cavity, and the other a rectangle of about an inch or so by half an inch 4. A thin coating of OCT is put on the bottom of the mold to act as glue, so the tissue won?t pop out during freezing. 5. A thin walled aluminum cup----2.75 to 3 inches diameter at the brim and slightly narrower at the bottom (mine was meant to be a gravy mixer) is filled with a cup or more of isopentane and lowered into the liquid nitrogen. The cup is best held with long forceps. When the fizzing stops for the second time---there is usually a secondary roiling of the isopentane----a SMALL bit of liquid nitrogen is admitted into the cup ( No need my telling you a small bit---use more---you won?t want to do it twice---it boils all over the place! ) and stirred. I used a butter knife with thick wrapping of paper towels around the handle---it gets COLD. This is done until you have. a thick slurry. The muscle in the mold is quickly put into the slurry and dunked to a slow count of 10 to 20. The mold is lifted out of the slurry, shaken twice quickly, to shake off the clinging isopentane, and put in the liquid nitrogen for 10- to 20 slow dunks. Then the mold is put directly into the cryostat. 6. The tissues are removed from the mold inside the cryostat. The frozen OCT with the muscle in it is placed on a specimen holder for your cryostat. It is then bolstered about with OCT ---the bolstering results in a neat rectangle wider across than up and down. All of the edges are trimmed to be straight and clean---a straight edged, square cornered rectangle which is cut with the broad bottom to the knife. You will see that with edges straight and neat, you will have happier time cutting sections. Go watch the residents sometimes, cutting frozens with the bare tissue turned any which way, splitting, turning, getting cussed. 7. Sections are put onto 2 by 3 inch glass slides. Using cover glass is a volunteering for misery. I can?t imagine how that got started. You can handle and store the glass slides easier and 178.56 times safer than when on cover glass. 8. Reagents are mixed to FILL the coplin jars, completely covering the tissues on the slides. Stingy volumes of histochemical preparations is inherited thru the genes from Scrooge!! 9. Sigma sells a buffer called 221 that you will see used in the ATP-ase and the other preparations in the packet. . It does not require a controlled substance license to purchase and it beats the socks off the sodium pentobarbital in all functions. All of this is better covered in the video. Now then, what?s going to happen? What will win out there in your labs----habit or function?? -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031015/2966f830/attachment.htm From Rcartun <@t> harthosp.org Wed Oct 15 12:46:31 2003 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Histology Supervisor Message-ID: Are there any Histology Laboratories out there with over 40,000 surgical pathology accessions that do not have a full-time supervisor? Thanks! Richard Cartun From bill501 <@t> mindspring.com Wed Oct 15 13:36:11 2003 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Flatbed Scanners for EM negatives In-Reply-To: <3F8D82FE.10401@ucsd.edu> References: <200310150914.AA104464676@popmail.opt.uh.edu> <3F8D82FE.10401@ucsd.edu> Message-ID: At 10:25 AM -0700 10/15/03, K. Bowden wrote: >We use Macintosh computers and do most all of our scanning on an >AGFA Arcus II. I'm not sure if they sell that model any longer but >I'm sure the latest model would have at lest the same quality if not >better. We scan x-rays, 35 mm slides, we have even placed MMA >embedded sections directly in the scanner and got excellent results. Do you scan directly on the scanner glass or does the scanner have a negative attachment or cover? I use UMAX Powerlook III's and get poor results when using the normal cover. But, with the negative attachment (which is a different cover) works fine. The light is transmitted through the negative and the CCD sensor is in the cover which picks up the image. Bill -- ______________ Bill Blank, MD Heartland Lab, Inc From kbowden <@t> ucsd.edu Wed Oct 15 14:19:09 2003 From: kbowden <@t> ucsd.edu (K. Bowden) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Flatbed Scanners for EM negatives References: <200310150914.AA104464676@popmail.opt.uh.edu> <3F8D82FE.10401@ucsd.edu> Message-ID: <3F8D9DAD.2050405@ucsd.edu> When we bought the Arcus II they advertised that you could scan 35mm slides. It had the highest resolution at the time and is 36 bit color. Now you can get even higher resolution and 48 bit. It only came with one cover which has the light in it. So there is two light sources one in the cover and one in the base. You change the setting in the software to choose between reflective or transparent. We liked the idea of not having to change covers. We thought it was safer for the scanner to not have someone possibly drop a cover while changing it. The only problem we have is for that model they don't have a driver that works with OS X so we have to use it with one of our older computers. I personally own a epson scanner and I like the AGFA better. -- Karen Bowden Staff Research Associate II University of CA, San Diego Department of Orthopedics 9500 Gilman Dr. 0630 La Jolla, CA 92093-0630 858-277-4970 CONFIDENTIALITY NOTICE: 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. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVED THIS E-MAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER. Bill Blank wrote: > At 10:25 AM -0700 10/15/03, K. Bowden wrote: > >> We use Macintosh computers and do most all of our scanning on an AGFA >> Arcus II. I'm not sure if they sell that model any longer but I'm >> sure the latest model would have at lest the same quality if not >> better. We scan x-rays, 35 mm slides, we have even placed MMA >> embedded sections directly in the scanner and got excellent results. > > > Do you scan directly on the scanner glass or does the scanner have a > negative attachment or cover? I use UMAX Powerlook III's and get poor > results when using the normal cover. But, with the negative attachment > (which is a different cover) works fine. The light is transmitted > through the negative and the CCD sensor is in the cover which picks up > the image. > > Bill From michael_lafriniere <@t> memorial.org Wed Oct 15 14:34:45 2003 From: michael_lafriniere <@t> memorial.org (LaFriniere, Michael) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Histology Supervisor Message-ID: Richard, I'm not sure if this answers your question completely but may help. The NSH Productivity Task force recent findings found that most pathology labs with 25,000 surgical specimens and higher employ a "non bench" supervisor/manager. Michael LaFriniere, PA, HT(ASCP) NSH Region III Director Chair NSH Productivity Task Force Pathology Manager Memorial Hospital Chattanooga TN 423-495-6117 -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Wednesday, October 15, 2003 1:47 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histology Supervisor Are there any Histology Laboratories out there with over 40,000 surgical pathology accessions that do not have a full-time supervisor? Thanks! Richard Cartun _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kgrobert <@t> rci.rutgers.edu Wed Oct 15 15:32:13 2003 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Flatbed Scanners for EM negatives In-Reply-To: <200310150914.AA104464676@popmail.opt.uh.edu> References: <200310150914.AA104464676@popmail.opt.uh.edu> Message-ID: <3F8DAECD.6050001@rci.rutgers.edu> Well, we have a Minolta Dimage Scan Multi II film scanner, but so far I have been unable to find out who offers the large-format EM film holder for it (it was mentioned in the description of the scanner on the Web site of the company that sold it to me, but nobody there seemed to know where to get the darn thing). I even emailed Minolta, but nobody got back to me about it. This does very well with 35 mm film negatives and slides, for what it is worth. However, for the EM film we have a HP Scanjet 5370C with transparency adapter that is working very well for us. Unfortunately, HP has discontinued this model, but I managed to find one by doing a Web search. HP may have a similar model to replace it, though, so I would start with hp.com and go from there. Good luck, Kathleen Roberts Principal Lab Technician Neurotoxicology Laboratories Rutgers University Margaret Gondo wrote: >Hi all - > >I was just wondering if anyone out in Histoland has any >recommendations on flatbed scanners that will do a decent job of >scanning an EM negative. Our lab uses mostly Macintosh computers >for imaging. I will admit that I know very little about scanners. >I just visited a site that had film scanners that cost in the >thousands along with flatbed scanners with similar DPI for a couple >hundred. > >Any information would be greatly appreciated. > >Margaret > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From khbarr <@t> mdanderson.org Wed Oct 15 15:56:29 2003 From: khbarr <@t> mdanderson.org (khbarr@mdanderson.org) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Available Positions Message-ID: The University of Texas M. D. Anderson Cancer Center is in need of two Histology Technicians in the Main Histology Laboratory (routine, processing, embeddin, cutting, staining) and one Sr. Histology Technician in the Special Microtomy/Bone Laboratory (embedding, cutting, staining bone marrow biopsies/clots, bone specimens, recuts, unstained slides). We are also seeking a Pathologist Assistant. Interested candidates may contact Dianna Menard at 713-745-6184, email her at dmenard@mdanderson.org or apply on line at www.mdanderson.org. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031015/5265573f/attachment.htm From garygill <@t> dcla.com Wed Oct 15 15:37:26 2003 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] ASR listing Message-ID: Can anyone point me to a comprehensive listing of immunohistochemistry reagents and their status as ASRs, IVDs, RUOs, and IUOs, and which ones require the ASR disclaimer? Thanks. Gary Gill From csawrenk <@t> bccancer.bc.ca Wed Oct 15 16:33:51 2003 From: csawrenk <@t> bccancer.bc.ca (Christina Sawrenko) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Albumin probe; Fli-1 Message-ID: <6BAF4D075F07D411B30900508B94CBA09C6D20@SERVER20> Hello Histonetters, We would like to know if anyone has info on an albumin probe for in-situ hybridization and if anyone has used the antibody Fli-1. Regarding Fli-1, where are you getting the antibody and could you share your protocol? Thanks in advance for your help! Chris Sawrenko Histopathology BC Cancer Agency Vancouver, British Columbia -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031015/d1a4fc7d/attachment.htm From JMahoney <@t> alegent.org Wed Oct 15 16:33:53 2003 From: JMahoney <@t> alegent.org (Janice A Mahoney) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] ASR listing Message-ID: Gary, There is no list. It depends on the manufacturer labeling and how each lab is using each antibody. Jan, Omaha >>> Gary Gill 10/15/2003 3:37:26 PM >>> Can anyone point me to a comprehensive listing of immunohistochemistry reagents and their status as ASRs, IVDs, RUOs, and IUOs, and which ones require the ASR disclaimer? Thanks. Gary Gill _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JGordon <@t> cellmarque.com Wed Oct 15 16:37:01 2003 From: JGordon <@t> cellmarque.com (Jeff Gordon) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] ASR listing Message-ID: Gary, each of Cell Marque's antibodies are listed individually as either an IVD or an ASR (we don't carry any RUOs). To make it easy for you, every one of Cell Marque's antibodies are IN VITRO DIAGNOSTIC antibodies with the following exceptions: Adenovirus CMV EBV Hepatitis B Core Antigen Hepatitis B Surface Antigen HSV 1&2 Parvovirus B19 Pneumocystis Carinii Her2/NU ER PR Ki-67 p53 Basically, with our antibodies it comes down to all of the viral markers being ASRs, pneumocystis being an ASR, and the breast prognostic markers being ASRs. As far as I know, ALL ASRs require a disclaimer when used in a clinical lab. The rest that we carry so far (all the CD markers, all the keratins, all the hormonal markers, all the melanoma markers, etc.) are IVDs. Other companies, depending on their use of each marker, may have different designations for their antibodies. Unfortunately, I don't believe that there is a universal list for antibodies from all companies that lists whether they are IVD, ASR, or RUO. Since we are geared primarily towards clinical labs, we carry antibodies approved for use in clinical settings. Some companies may have the same antibodies (even the same clones) geared towards research labs, and theirs may be designated as Research Use Only. Jeff Gordon Cell Marque Corp. 1-800-665-7284, Ext. 12 -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Wednesday, October 15, 2003 3:37 PM To: 'Histonet' Subject: [Histonet] ASR listing Can anyone point me to a comprehensive listing of immunohistochemistry reagents and their status as ASRs, IVDs, RUOs, and IUOs, and which ones require the ASR disclaimer? Thanks. Gary Gill _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Wed Oct 15 17:29:22 2003 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Flatbed Scanners for EM negatives Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E062@simba.kids> Margaret, Last month I posted the following email. I have uploaded pictures to the Histonet pics page (http://www.histonet.org/site_images_frame.asp) 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 Subject: RE: [Histonet] Scanner for microscope slides We use an Epson Perfection 2450 Photo Scanner. We place the glass slide in the 35mm holder, place it on the scanner (ensure that back light cover is off) and scan as you would a 35mm slide. We also use the scanner for EM negatives, 35mm photos and picture scanning for Grand Rounds etc. -----Original Message----- From: Margaret Gondo [mailto:MGondo@popmail.opt.uh.edu] Sent: Thursday, 16 October 2003 0:15 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Flatbed Scanners for EM negatives Hi all - I was just wondering if anyone out in Histoland has any recommendations on flatbed scanners that will do a decent job of scanning an EM negative. Our lab uses mostly Macintosh computers for imaging. I will admit that I know very little about scanners. I just visited a site that had film scanners that cost in the thousands along with flatbed scanners with similar DPI for a couple hundred. Any information would be greatly appreciated. Margaret _______________________________________________ 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 Wed Oct 15 16:01:00 2003 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Flatbed Scanners for EM negatives In-Reply-To: <3F8D9DAD.2050405@ucsd.edu> References: <200310150914.AA104464676@popmail.opt.uh.edu> <3F8D82FE.10401@ucsd.edu> <3F8D9DAD.2050405@ucsd.edu> Message-ID: At 12:19 PM -0700 10/15/03, K. Bowden wrote: >The only problem we have is for that model they don't have a driver >that works with OS X so we have to use it with one of our older >computers. Have you trued Vuescan? My UMAX scanner don;t have OS X drivers either, but Vuescant works very well in OS X. -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) From katri <@t> cogeco.ca Wed Oct 15 20:50:27 2003 From: katri <@t> cogeco.ca ( Katri Tuomala) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] AEC References: <3303.193.255.128.130.1066220238.webmail@mail1.netbulmail.com> Message-ID: <003d01c39387$e06836a0$ce989618@hala4.on.cogeco.ca> Izzet, AEC is soluble in organic solvents, so you need to coverslip from water or a buffer with a water based mounting medium. Many are commercially available or you can make your own with glycerin. Katri Katri Tuomala St. Joseph's Healtcare Hamilton, Ontario, Canada ----- Original Message ----- From: "izzet oguz" To: Sent: Wednesday, October 15, 2003 8:17 AM Subject: [Histonet] AEC > Hi there, > we all used DAB for IHC before..but now we have an AEC kit and we suppose > that dehidration isn't used for AEC but what about xylol?..do we put the > slides in xylol after counterstain or not? Is there any difference apart > from that..Will you please share your experiences?.. > Thanks > ?zzet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From m.vermehren <@t> anat.vetmed.uni-muenchen.de Thu Oct 16 02:58:22 2003 From: m.vermehren <@t> anat.vetmed.uni-muenchen.de (Margarete Vermehren) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] immunohistochemistry on non adherent cells Message-ID: <000c01c393bb$45ac6800$64ae548d@HistoPC3> This should work for a confocal, so I cannot embed the pellet in paraffin... Can anybody share a protocol with me? Thank you very much for advice! Margaret Margarete Vermehren LMU M?nchen / Tieranatomie II Tel: +49 89 21805857 Fax: +49 89 21802569 m.vermehren@anat.vetmed.uni-muenchen.de From Jim.Tunstead <@t> msnyuhealth.org Thu Oct 16 09:58:14 2003 From: Jim.Tunstead <@t> msnyuhealth.org (Tunstead, Jim) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Histotechnologist position in New York City Message-ID: <1371F1DB5EE3294CA931D4321EBCB0041A9F85@EXCEBW2K18.msnyuhealth.org> Dear All, We currently have a position available at Mount Sinai School of Medicine which may be of interest to list users. Please see details below. Histotechnologist / Lab Coordinator We currently have an opening for a Histotechnologist. The individual will be an integral part of a newly equipped pathology core laboratory. The core provides histology, immunohistochemistry and image analysis support for the Cardiovascular Institute. Services are provided to a small group of investigators, allowing greater involvement in their research projects. As well as advising on general procedures, the successful candidate will have the opportunity to develop new processing and staining protocols. Specific Responsibilities: Act as a histology and immunochemistry resource for researchers in the preparation of their protocols. Assist in establishing, optimizing, customizing and documenting protocols for fixing, processing, sectioning, staining, and reagent preparation for research projects. Actively participate in group meetings with researchers to discuss experimental design. Fix, process and paraffin embed samples; process and freeze fresh samples. Perform sectioning of paraffin and frozen blocks for immunohistochemistry, in situ hybridization and special stains. Accurately label, organize, and track tissue, blocks, and slides. Prepare reagents from detailed laboratory procedures and document that quality control standards are being met. Research, troubleshoot, and test new reagents. Inventory, maintain, and order supplies as necessary. Assist in establishing fees and processing billing. Perform routine maintenance on laboratory equipment, including automated immunostainer, microtome, cryostat, freezer, and tissue processor. Assist in organizing work and storage space, and assist in general lab maintenance. Perform other duties, as assigned. The ideal candidate will possess the following qualifications: The minimum requirement is a high school diploma/AA with formal or on the job training as a histology technician. A Bachelor of Science degree and HT/HTL certification by a nationally recognized certifying agency, such as the American Society of Clinical Pathology, are desirable but not required. A minimum of three years of experience is required and good microtome sectioning skills are essential. Additional experience and training are highly desirable, particularly in a research setting. Experience in the use of an automated immunohistology stainer, clinical and research histology experience, and a strong scientific background are very desirable. Experience with microscopy, tissue procurement, and non-human histology is also a plus. The successful technician must be able to organize and prioritize tasks, work accurately and efficiently, be detail oriented, work both independently and with a group, and have excellent written and verbal communication skills. Familiarity with Microsoft Word, Excel, and Internet Explorer is desirable. Business Title: Lab Coordinator Job Type: Full time Job Category: Laboratory research Compensation: Salary DOE + excellent benefits CLOSING DATE: Open until filled. If you have questions or wish to apply, please contact Jim Tunstead by e-mail, phone, fax or mail. Jim Tunstead, Ph.D. Assistant Professor Cardiovascular Institute, Box 1030 Mount Sinai School of Medicine One Gustave Levy Place New York, NY 10029 ph:212-241-0731 (Jacinta Russell) fx: 212-860-7032 jim.tunstead@msnyuhealth.org Jim Tunstead, Ph.D. Assistant Professor Cardiovascular Institute, Box 1030 Mount Sinai School of Medicine One Gustave Levy Place New York, NY 10029 ph:212-241-0731 (Jacinta Russell) fx: 212-860-7032 jim.tunstead@msnyuhealth.org From alust1 <@t> hotmail.com Thu Oct 16 09:22:46 2003 From: alust1 <@t> hotmail.com (Andrew Lust) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] postive control slides collagen type III Message-ID: Dear Histonetters I am an undergrad at the University of Houston, in need of positive controls slides of Collagen type 3. If anyone is willing to spare a slide or two to a struggling undergrad please respond to this email. or You can contact me at Alust1@hotmail.com. Thanks again Andrew T. Lust _________________________________________________________________ Add MSN 8 Internet Software to your current Internet access and enjoy patented spam control and more. Get two months FREE! http://join.msn.com/?page=dept/byoa From HornHV <@t> archildrens.org Thu Oct 16 09:42:56 2003 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] ASR listing Message-ID: You will find that information on your product inserts from whomever you buy your antibodies. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: Gary Gill [mailto:garygill@dcla.com] Sent: Wednesday, October 15, 2003 3:37 PM To: 'Histonet' Subject: [Histonet] ASR listing Can anyone point me to a comprehensive listing of immunohistochemistry reagents and their status as ASRs, IVDs, RUOs, and IUOs, and which ones require the ASR disclaimer? Thanks. Gary Gill _______________________________________________ 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. Arkansas Children's Hospital From millicent_renee <@t> yahoo.com Thu Oct 16 10:46:33 2003 From: millicent_renee <@t> yahoo.com (Millicent Bruce) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] burned artifact Message-ID: <20031016154633.71086.qmail@web12307.mail.yahoo.com> For the past couple of years we have been getting a burned artifact in our biopsies. The pathologist have said that on the slides the nucleus look smudged or burned. It is mostly on the small bxs, but we have seen it on larger specimens also. We have bought a new tissue processor, and a new slide stainer thinking that might be the problem. We are still seeing the artifact. Is there anybody that might can tell us what is causing the problem? We have tried everything we can think of. __________________________________ Do you Yahoo!? The New Yahoo! Shopping - with improved product search http://shopping.yahoo.com From mward <@t> wfubmc.edu Thu Oct 16 10:59:41 2003 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Myogenin Message-ID: <61135F0455D33347B5AAE209B903A304032622CB@EXCHVS2.medctr.ad.wfubmc.edu> I was wondering what vendor folks are using for myogenin? We have both a Ventana NexEs and Dako autostainer and am looking for the best method and vendor. We are getting off and on staining. Any help is appreciated. Thanks! Martha Ward Wake Forest University Baptist Medical Center From ASelf <@t> gmhsc.com Thu Oct 16 11:11:59 2003 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Chain of Custody for LEA Message-ID: Hello HistoNetters, Does anyone have a policy/procedure on how they handle specimens being removed from the body for Law Enforcement Agencies(LEA) that they could share with me. We have a Chain of Custody form that we fill out for such incidents but we need to write up a procedure. My doc was wandering how other hospitals were handling these situations whether the specimen/foreign body be removed in the OR, ER, or from an autopsy. Thanks for your help in advance... Amy Self Georgetown Memorial Hospital 843-520-7882 (fax) Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or 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 Loralee_Gehan <@t> URMC.Rochester.edu Thu Oct 16 12:35:50 2003 From: Loralee_Gehan <@t> URMC.Rochester.edu (Gehan, Loralee) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] question about frozen sections Message-ID: <95774A6A6036D411AFEA00D0B73C86430888045D@exmc3.urmc.rochester.edu> I am currently cutting frozen sections of formaldhyde fix mouse legs. What is the procedure for storage of these slides? Do they need to be stored in a freezer or can I just leave them at room temp. Thanks in advance. Loralee Gehan Orthopaedics Research Lab University of Rochester Rochester, NY From laurie.colbert <@t> huntingtonhospital.com Thu Oct 16 13:10:23 2003 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Chain of Custody for LEA Message-ID: <0BE6ADFAE4E7E04496BF21ABD34662801D0C5A@EXCHANGE1.huntingtonhospital.com> Amy, I would be interested in any info you get, as I need to write a procedure, too. Right now, Surgery hand-carries the specimen (usually a bullet) to Pathology (or locks it up until Pathology is open)and everyone that has handled the specimen has to sign the Chain of Custody form. Our pathologist grosses it and then we lock it up until a detective comes to get it. A lot of times we have problems with aliases, as the name the detective has is not the same name that we have. What I would like to see is our Security department to get involved. I would like them to pick up the specimen from Surgery, bring it to Pathology and wait while a Pathologist grosses it, and then take it for safe-keeping. When the detectives come for the specimen, they will contact Security. Laurie Colbert Huntington Memorial Hospital Pasadena, CA -----Original Message----- From: Amy Self [mailto:ASelf@gmhsc.com] Sent: Thursday, October 16, 2003 9:12 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Chain of Custody for LEA Hello HistoNetters, Does anyone have a policy/procedure on how they handle specimens being removed from the body for Law Enforcement Agencies(LEA) that they could share with me. We have a Chain of Custody form that we fill out for such incidents but we need to write up a procedure. My doc was wandering how other hospitals were handling these situations whether the specimen/foreign body be removed in the OR, ER, or from an autopsy. Thanks for your help in advance... Amy Self Georgetown Memorial Hospital 843-520-7882 (fax) Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or 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 CCLYATT <@t> mail.mcg.edu Thu Oct 16 13:18:22 2003 From: CCLYATT <@t> mail.mcg.edu (Claye Clyatt) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] burned artifact Message-ID: Check to see if the submitting department is using any type of cautery instrument in the collecting process. Are the biopsies placed IMMEDIATELY into the fixative? Eith of these circumstances will produce an artifact on your slides. Claye Claye Clyatt Chief Histotechnologist Department of Pathology Room #BF119 Medical College of Georgia Augusta, Ga 30912 office (706) 721-3630 pager (706) 721-7243-1132 e-mail: cclyatt@mail.mcg.edu >>> Millicent Bruce 10/16/03 11:46AM >>> For the past couple of years we have been getting a burned artifact in our biopsies. The pathologist have said that on the slides the nucleus look smudged or burned. It is mostly on the small bxs, but we have seen it on larger specimens also. We have bought a new tissue processor, and a new slide stainer thinking that might be the problem. We are still seeing the artifact. Is there anybody that might can tell us what is causing the problem? We have tried everything we can think of. __________________________________ Do you Yahoo!? The New Yahoo! Shopping - with improved product search http://shopping.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Annette_hall <@t> pa-ucl.com Thu Oct 16 14:18:17 2003 From: Annette_hall <@t> pa-ucl.com (Annette Hall) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Stainer & Coverslipper thoughts Message-ID: <21EE03EF1DB9D511B112000255FC299486E058@mercury.pa-ucl.com> Having successfully implemented the Artisan by DakoCytomation, we are looking to introduce more automation to our histology department. An H&E stainer and coverslipper seem the next logical purchases. Can anyone share their experiences, good or bad, with stainers and coverslippers they have used? I realize your time is precious and I thank anyone for their valuable input. Annette J Hall, MT Micro/Histo/Cyto Supervisor United Clinical Labs 205 Bluff St. Dubuque, IA 52001 563.556.2010 x131 From MTitford <@t> aol.com Thu Oct 16 14:20:15 2003 From: MTitford <@t> aol.com (MTitford@aol.com) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Chain of custody Message-ID: <5184878F.1E4A0F04.00762DB1@aol.com> Amy Self & Laurie Colbert ask about chain of ownership on bullets. We have bullet boxes in our operating room, and in the emergency department. They are like mail boxes with a slot on top but they lock.(You can place stuff in it, but not get it out!) Nurses place the bullet in a labelled plastic bag with a copy of the surgical pathology request form and place it in the bullet box,and log the specimen in on a special log. On the next working day our gross room tech unlocks the box (we have the only key) and signs for the bullet. After the bullet is grossed in, we lock it away again. Sometimes, a long time later, the bullet gets subpoenaed We have our risk management department handle subpoenas, but we occasionally get calls from police departments asking if we have a bullet from such and such a patient. Because of HIPPA we have to tell them to subpoena the record. Our University lawyer has blessed this procedure. Mike Titford USA Pathology Mobile AL From jmacdona <@t> mtsac.edu Thu Oct 16 13:59:53 2003 From: jmacdona <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] postive control slides collagen type III References: Message-ID: <001301c39417$aed57810$7934908c@JMACDONADT> Reticular fibers are collagen, type III. Liver is a good control for reticular fibers, as is spleen. Jennifer MacDonald ----- Original Message ----- From: "Andrew Lust" To: Sent: Thursday, October 16, 2003 7:22 AM Subject: [Histonet] postive control slides collagen type III > Dear Histonetters > > I am an undergrad at the University of Houston, in need of positive controls > slides of Collagen type 3. If anyone is willing to spare a slide or two to > a struggling undergrad please respond to this email. or You can contact me > at Alust1@hotmail.com. > > Thanks again > Andrew T. Lust > > _________________________________________________________________ > Add MSN 8 Internet Software to your current Internet access and enjoy > patented spam control and more. Get two months FREE! > http://join.msn.com/?page=dept/byoa > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tissuearray <@t> hotmail.com Thu Oct 16 15:02:24 2003 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Fwd: Tissue Microarrays and the Block Warmer Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031016/6f4b6879/attachment.htm From mbryhan <@t> NORTHERNHEALTH.ORG Thu Oct 16 14:22:13 2003 From: mbryhan <@t> NORTHERNHEALTH.ORG (Mary Bryhan) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Cyclin D1 Message-ID: Dear Roy, I found out today through our purchasing department that Cyclin D1 has been discontinued by BioGenex. I called and spoke with Amanda in technical support who said that the antibody had problems and was pulled. This is not the first time that an antibody, or kit constituent has been identified as "bad" without any notice to us, the customer. If we are to keep our working relationship, I expect that BioGenex will notify us as soon as a problem is detected. Cordially, Mary Bryhan From tpmorken <@t> labvision.com Thu Oct 16 14:37:33 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] See you at NSH! Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CC21@usca0082k03.rallansci.apogent.com> For those going to NSH, hope to see you there! Tim Morken Product Development Lab Vision / NeoMarkers 47790 Westinghouse Dr Fremont, CA 94539 PH: 510-991-2840 www.labvision.com -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031016/4744df1e/attachment.htm From ohenry <@t> dfw.net Thu Oct 16 15:52:07 2003 From: ohenry <@t> dfw.net (Susan Owens) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] What is Hollande's fixative? Message-ID: <000901c39427$73aa93a0$5bdd3040@Nationwide.net> A few weeks ago I posted to Histonet and mentioned Hollande's fixative. As a result I rec'd a couple of private posts asking what was Hollande's fixative, they had never heard of it. So below I give you the formula for Hollande's and a quote from Freida Carson's book "Histotechnology A Self-Instructional Text". HOLLANDE'S SOLUTION/FIXATIVE Copper acetate........................... 25. gms Picric acid................................... 40. gms Formaldehyde,37% to 40%.......... 100. mls Acetic acid.................................. 15. mls Distilled water............................. 1,000. mls QUOTE: "This modification of Bouin's solutions is stable and will decalcify small specimens of done. It is becoming widely used as a fixative for biopsy specimens of the gastrointestinal tract. Hollande's-fixed tissue can be stained successfully with most special stains. The cupric acetate present in the solution stabilizes red blood cell membranes and the granules of eosinophils and endocrine cells, so that less lysis occurs than with Bouins solution. The fixative must be washed out before the specimen is placed in a phosphate-buffered formalin solution on the tissue processor; salts present in the solution will form an insoluble phosphate precipitate." end quote We buy ours from Stat Lab in prefilled bx containers. Hope this answers those questions and I hope Ms.Carson doesn't mine the quote. Susan Susan Owens-TX ohenry@dfw.net fax: 817-548-9876 "A bad day at the dog show is better then a good day at work!" From Tiffany.L.Sheffield <@t> uth.tmc.edu Thu Oct 16 15:14:49 2003 From: Tiffany.L.Sheffield <@t> uth.tmc.edu (Tiffany L Sheffield) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] See you at NSH! References: <8CD0CDC791DCC343ABB1F15298E4F3C417CC21@usca0082k03.rallansci.apogent.com> Message-ID: <3F8EFC39.B50D3048@uth.tmc.edu> Skipped content of type multipart/alternative-------------- next part -------------- A non-text attachment was scrubbed... Name: Tiffany.L.Sheffield.vcf Type: text/x-vcard Size: 374 bytes Desc: Card for Tiffany Sheffield-Lopez Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031016/6fd883cd/Tiffany.L.Sheffield.vcf From lesley <@t> vancouverbc.net Thu Oct 16 17:01:41 2003 From: lesley <@t> vancouverbc.net (Lesley Weston) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Tetracycline and decalcification Message-ID: Does anyone know if decalcifying tetracycline-treated bone affects its fluorescence? Everything I can find on the subject says that they embedded undecalcified bone in methacrylate, but is this essential? If it is OK to decalcify, should one use acid or EDTA? Sorry to be so helpless, but this doesn't seem to have been covered anywhere. Thanks to anyone who can help. Lesley Weston. From AnthonyH <@t> chw.edu.au Thu Oct 16 18:00:11 2003 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] burned artifact Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E066@simba.kids> Bruce, My suggestions are: Don't let the fixation in NBF be too short. Use a more gentle processing (less pressure & vacuum, lower temperature, shorter time in alcohols and xylene). 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 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html -----Original Message----- From: Millicent Bruce [mailto:millicent_renee@yahoo.com] Sent: Friday, 17 October 2003 1:47 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] burned artifact For the past couple of years we have been getting a burned artifact in our biopsies. The pathologist have said that on the slides the nucleus look smudged or burned. It is mostly on the small bxs, but we have seen it on larger specimens also. We have bought a new tissue processor, and a new slide stainer thinking that might be the problem. We are still seeing the artifact. Is there anybody that might can tell us what is causing the problem? We have tried everything we can think of. __________________________________ Do you Yahoo!? The New Yahoo! Shopping - with improved product search http://shopping.yahoo.com _______________________________________________ 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 nheuer <@t> anhb.uwa.edu.au Thu Oct 16 21:55:15 2003 From: nheuer <@t> anhb.uwa.edu.au (Heuer N) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Cutting undecalcified rat teeth Message-ID: I am a Medical Scientist having majored in Histology, however my studies have not given me the skills to tackle a project i am involved in. I have been requested to section undecalcified rat teeth for LM, and have no clue as to how to approach this matter. i am wondering if anyone had any information that would be helpful ( ie. websites or journal articles i could source ...anything??). Any guides in the right direction would be really appreciated Natascha Heuer The University of Western Australia School of Anatomy & Human Biology 35 Stirling Hwy Crawley WA 6009 Ph: (08) 9380 3798 Fax: (08) 9380 1051 Email: nheuer@anhb.uwa.edu.au From nick.kirk3 <@t> btopenworld.com Fri Oct 17 03:12:54 2003 From: nick.kirk3 <@t> btopenworld.com (nick.kirk3@btopenworld.com) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Stainer & Coverslipper thoughts Message-ID: <5490429.1066378374105.JavaMail.root@127.0.0.1> It's worth having a look at the Leica ST5020 robotic stainer with intergrated CV5030 coverslipper. Basically you put sections on one end and get stained and coverslipped slides at the other. It saves a lot of time, reduces exposure to Xylene and is quick and efficient. I also like the touch screen control system. Nick Kirk Head Biomedical Scientist Histopathology Hinchingbrooke Hospital Huntingdon England > from: Annette Hall > date: Thu, 16 Oct 2003 20:18:17 > to: histonet@lists.utsouthwestern.edu > subject: Re: [Histonet] Stainer & Coverslipper thoughts > > > Having successfully implemented the Artisan by DakoCytomation, we are > looking to introduce more automation to our histology department. An H&E > stainer and coverslipper seem the next logical purchases. Can anyone share > their experiences, good or bad, with stainers and coverslippers they have > used? I realize your time is precious and I thank anyone for their valuable > input. > > Annette J Hall, MT > Micro/Histo/Cyto Supervisor > United Clinical Labs > 205 Bluff St. > Dubuque, IA 52001 > 563.556.2010 x131 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From doscwk <@t> nus.edu.sg Fri Oct 17 03:52:38 2003 From: doscwk <@t> nus.edu.sg (Chan Wai Kam) Date: Fri Sep 16 15:22:03 2005 Subject: [Histonet] Auto stainer and coverslipper Message-ID: Hi Histonetters, I would like to find out the quality of staining and coverslipping done on those auto equipment. We still stain our slides manually. I'm just wondering whether those slides coverslipped by machine would result in any slides with air bubbles in them (as we sometimes get with manual coverslipping). Thanks Julee Chan Orthopaedic Surgery National University of Singapore From DDDeltour <@t> sig.med.navy.mil Fri Oct 17 04:14:09 2003 From: DDDeltour <@t> sig.med.navy.mil (Deltour, Douglas D.(HM2)) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Auto stainer and coverslipper Message-ID: The consistency is the nice part of auto stainers. I would recommend a cover slipper depending on the quantity of slides produced. HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy LPO Histology Histology Technician DSN 624-4669 FAX 624-4680 COMM (From US) 011-39-095-56-4669 DISCLAIMER: 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. **** Informazione Confidenziale - Questa e-Mail, incluso eventuali allegati, contiene informazioni confidenziali intese unicamente alla persona/e a cui e' indirizzata. Se avete ricevuto per errore questo messaggio, cortesemente contattare immediatamente il mittente e cancellare la e-Mail. Grazie****. -----Original Message----- From: Chan Wai Kam [mailto:doscwk@nus.edu.sg] Sent: Friday, October 17, 2003 10:53 AM To: HistoNet Server Subject: [Histonet] Auto stainer and coverslipper Hi Histonetters, I would like to find out the quality of staining and coverslipping done on those auto equipment. We still stain our slides manually. I'm just wondering whether those slides coverslipped by machine would result in any slides with air bubbles in them (as we sometimes get with manual coverslipping). Thanks Julee Chan Orthopaedic Surgery National University of Singapore _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nick.kirk3 <@t> btopenworld.com Fri Oct 17 05:00:07 2003 From: nick.kirk3 <@t> btopenworld.com (nick.kirk3@btopenworld.com) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Auto stainer and coverslipper Message-ID: <3767630.1066384807781.JavaMail.root@127.0.0.1> You can adjust the volume of mountant dispensed on all the automatic coverslippers that I've tried, which ensures adequate cover of the section and in my experience you get a lot less air bubbles than you do with manual methods. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England > from: Chan Wai Kam > date: Fri, 17 Oct 2003 09:52:38 > to: histonet@lists.utsouthwestern.edu > subject: Re: [Histonet] Auto stainer and coverslipper > > Hi Histonetters, > > I would like to find out the quality of staining and coverslipping done > on those auto equipment. We still stain our slides manually. I'm just > wondering whether those slides coverslipped by machine would result in > any slides with air bubbles in them (as we sometimes get with manual > coverslipping). > > Thanks > Julee Chan > Orthopaedic Surgery > National University of Singapore > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From georgecole <@t> ev1.net Fri Oct 17 06:56:07 2003 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] bubbles in cover slipping Message-ID: <000001c394a5$a741fc20$084dbad0@hppav> Folks ABOUT COVER GLASS AND BUBBLES--- Lee Luna---some of you may not have known Lee---he was editor of the government histological manual---he came to visit us several times at Good Samaritan Hospital. He loved an ice cream bar we could get around here---called THAT'S IT, or something like that. He and I got off our diets a couple a time per visit with that. One visit, he showed us a way to cover slip slides that was easier, faster and practically bubble proof. FIRST OF ALL--- WIPING THE SLIDES WITH A TOWEL IS A SILLY TIC. THERE IS MORE LINT AND STUFF PUT ONTO THE SLIDES FROM A TOWEL THAN WOULD BE THERE FROM THE FINAL CLEARING AGENT. You take two or three slides out of the clearing agent--- it was xylene with us---one at a time----- You lay them on two or three clean paper towels----you can replace the paper towels any time they get too wet with xylene, but you can do a lot of slides without doing so. This was done under the hood of course. You simply lay the slides out on the paper towels. AH AH AH---PUT THAT HAND TOWEL AWAY!!!--- You put a streak of the cover glass mounting medium you use on the upper or lower edge of the slides----which ever is more comfortable to you. You will learn quickly how much mounting medium to put on the slides. You then lay the cover glass one by one on the slides---if you put the mounting medium on the upper edge of one slide---you lay the upper edge of the cover glass in the mounting medium on the upper edge of that slide and lower it, leading the mounting medium downward before it with no bubbles. If you do make bubbles, you can draw them down and off the lower edge of the slide. You then do the same for slides two and three. The ventilation in our lab was strong---I could never do more than three slides at a time this way----the mounting medium would be evaporating. This was quick and neat---and you could easily control bubbles. There are those who will insist on continuing to hold the slides up in the air in front of their faces to cover them----folks, if you do---you might as well cover slip while standing on your heads. Just breathe slowly, smile, and try the above. It's hygienic and low cal. If you put the mounting medium on the lower edge of the slide, you simply appliy the cover glass to the lower edge and lower it up to the top of the slide leading any bubbles off the edge of the slide. And, with either upper or lower edge application, it was easy to learn how to put the cover slip on without making bubbles. But if you did, you could scoot the bubbles off with ease when lowering the cover glass. There was a variation on this that some techs liked----you lay the COVER GLASSES out on the paper towels, one two or three----put the mounting medium on the upper or lower edges of each cover glass---than touch the lower edge of the slide itself over the cover glass in the mounting medium---the mounting medium will fill up the space naturally between the cover and the slide with few bubbles resulting----if you do make bubbles, you simply lead them off when you finish lowering the slide onto the cover glass---either from the bottom up or from the top down. With all of the above, you finish with touching the bottom of the slides to the paper towels to drain any excess mounting medium off the slides. NO CLOTH TOWELS HERE. There are those who will prefer to touch both top and bottom of the slides to the paper towels. This is ok, but may not be necessary. You can do your covering quickly---neat, clean and bubble-free. I know from the histonet, many of you have auto stainers, and some of you have auto cover glassing equipment. I never did. The equipment was ME. If you can get bubble free cover slipping from those mechanical slaves, good. But I can say that bubbles were not unusual with slides covered by motorized cover slipping. Like John Henry, I was ready to beat the machine with quantity and quality but we never got one. Shucks! There might have been a song written about it---- Georgecole@ev1.net -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031017/f08b4d9e/attachment.htm From juan.gutierrez <@t> christushealth.org Fri Oct 17 06:59:51 2003 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Stainer & Coverslipper thoughts Message-ID: Unfortunately the Leica ST5020 has not come out here in the states. My predecessor purchased one 18 months ago and we still haven't got it. In the meantime we have tested the Surgipath Tribune and decided to go with it instead. It doesn't connect with the coverslipper, but we have found that the Leica CV5030 to be very unreliable anyway. On the year and a half we've had it, they had to come work on it at least twice and I'm about to place a service call on it again. Good luck with your quest. Juan C. Gutierrez, HT(ASCP) -----Original Message----- From: nick.kirk3@btopenworld.com [mailto:nick.kirk3@btopenworld.com] Sent: Fri 10/17/2003 3:12 AM To: histonet@lists.utsouthwestern.edu Cc: Subject: Re: [Histonet] Stainer & Coverslipper thoughts It's worth having a look at the Leica ST5020 robotic stainer with intergrated CV5030 coverslipper. Basically you put sections on one end and get stained and coverslipped slides at the other. It saves a lot of time, reduces exposure to Xylene and is quick and efficient. I also like the touch screen control system. Nick Kirk Head Biomedical Scientist Histopathology Hinchingbrooke Hospital Huntingdon England > from: Annette Hall > date: Thu, 16 Oct 2003 20:18:17 > to: histonet@lists.utsouthwestern.edu > subject: Re: [Histonet] Stainer & Coverslipper thoughts > > > Having successfully implemented the Artisan by DakoCytomation, we are > looking to introduce more automation to our histology department. An H&E > stainer and coverslipper seem the next logical purchases. Can anyone share > their experiences, good or bad, with stainers and coverslippers they have > used? I realize your time is precious and I thank anyone for their valuable > input. > > Annette J Hall, MT > Micro/Histo/Cyto Supervisor > United Clinical Labs > 205 Bluff St. > Dubuque, IA 52001 > 563.556.2010 x131 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Linresearch <@t> aol.com Fri Oct 17 07:02:42 2003 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] (no subject) Message-ID: <14b.2571dc13.2cc13462@aol.com> Hello, I am stll sruggling with eNos on FFPE rodent tissues. Any advice would be appreciated. Lin -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031017/ee825f94/attachment.htm From RossS <@t> BaylorHealth.edu Fri Oct 17 07:16:59 2003 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Contact info for Ross Stapf Message-ID: I have heard that someone on histonet was looking for me. I've been off line for about a month. They finally got my work email up and running yesterday. This is my new contact info. The new job is going great. We have a CAP inspection in a month, so I've had to jump right in, but the folks here had most things ready. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 RossS@baylorhealth.edu 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 kemlo <@t> tiscali.co.uk Fri Oct 17 08:58:06 2003 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] burned artifact In-Reply-To: <20031016154633.71086.qmail@web12307.mail.yahoo.com> Message-ID: <000201c394b6$b1532200$23d5e150@KEMLOS> I just get the feeling that if you have fixed the tissues properly then there is 'relatively' little you can do to ruin it. I'm impressed that you can buy all that just for the problem; try looking at your fixative. Make sure its isotonic, the right pH and strength, and that you have fixed it for long enough. Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 01270 877625 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-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Millicent Bruce Sent: 16 October 2003 16:47 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] burned artifact For the past couple of years we have been getting a burned artifact in our biopsies. The pathologist have said that on the slides the nucleus look smudged or burned. It is mostly on the small bxs, but we have seen it on larger specimens also. We have bought a new tissue processor, and a new slide stainer thinking that might be the problem. We are still seeing the artifact. Is there anybody that might can tell us what is causing the problem? We have tried everything we can think of. __________________________________ Do you Yahoo!? The New Yahoo! Shopping - with improved product search http://shopping.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From laurie.colbert <@t> huntingtonhospital.com Fri Oct 17 09:55:10 2003 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Auto stainer and coverslipper Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5BC33@EXCHANGE1.huntingtonhospital.com> We have the Sakura Tissue Tek SCA coverslipper that utilizes the film to coverslip. I love it for our slides - it is very fast, easy to use, and relatively maintenance-free. However, I see that you are in Orthopaedic Surgery and I would not recommend the film coverslipper for bone specimens. If your tissue doesn't adhere to the slide well, you can get a funky cornflaking artifact. Laurie Colbert Huntington Hospital Pasadena, CA -----Original Message----- From: Chan Wai Kam [mailto:doscwk@nus.edu.sg] Sent: Friday, October 17, 2003 1:53 AM To: HistoNet Server Subject: [Histonet] Auto stainer and coverslipper Hi Histonetters, I would like to find out the quality of staining and coverslipping done on those auto equipment. We still stain our slides manually. I'm just wondering whether those slides coverslipped by machine would result in any slides with air bubbles in them (as we sometimes get with manual coverslipping). Thanks Julee Chan Orthopaedic Surgery National University of Singapore _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Fri Oct 17 12:41:07 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] fixative for insects In-Reply-To: <4.3.2.7.2.20031015104722.00cc80f0@algranth.inbox.email.arizona.edu> References: <4.3.2.7.2.20031015104722.00cc80f0@algranth.inbox.email.arizona.edu> Message-ID: <3F9029B3.2050607@umdnj.edu> A formalin - alcohol - acetic acid mix is often used for insects.There are many variations on the exact composition but Lillie (1976) recommends: "concentrated formalin" ( 37-40% formaldehyde) : 10 ml glacial acetic acid: 5ml 95% ethanol: 85 ml. Fix for up to 24 hours. Geoff Andrea Grantham wrote: > I'm going to be processing whole mosquitoes and I was wondering what > would be the best fixative to use...and processing schedule if anybody > has one. > > Thanks. > Andi > ..................................................................... > : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > : Sr. Research Specialist University of Arizona : > : (office: AHSC 4212) P.O. Box 245044 : > : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : > :...................................................................: > http://www.cba.arizona.edu/histology-lab.html > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- -- ********************************************** 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 Linresearch <@t> aol.com Fri Oct 17 14:01:35 2003 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:22:04 2005 Subject: Fwd: [Histonet] (eNos Ab Message-ID: <7f.3de4da86.2cc1968f@aol.com> -------------- next part -------------- An embedded message was scrubbed... From: Linresearch@aol.com Subject: [Histonet] (no subject) Date: Fri, 17 Oct 2003 08:02:42 EDT Size: 3618 Url: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031017/bd87422b/attachment.eml From pam <@t> ategra.com Fri Oct 17 14:20:21 2003 From: pam <@t> ategra.com (Pam Barker (extension 234)) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Histology Professionals -Latest Update Message-ID: Hello Histonetters , I am presently on a search for 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. These 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 supervisory, team lead and bench positions. These positions require HTL or HT certification or registry eligibility. Here are my newest openings: 1. Syracuse, NY - Histology Supervisor 2. Richmond, VA - Lead Histo Tech 3. Pittsburgh, PA - Histo Tech 4. Boston, MA - Histo Tech 5. Salt Lake City, UT - Histo Tech Here are some of my hottest Histology Supevisory positions: 1. Maine - Histology Supervisor 2. San Jose, CA - Histology Supervisor or Team Lead 3. Northern NJ - Histology Supervisor or Team Lead 4. Atlanta, GA - Team Lead grossing required 5. Illinois - Team Lead 6. New Hampshire - Histo Supervisor Here are some of my hottest Histo Tech bench positions: 1. Southwest FL - opportunity to obtain QIHC some grossing required 2. Orlando/Tampa - IHC and routine histology 3. Maine - routine histology 4. Louisiana - MOHS 5. Illinois - Team Lead 6. Boulder, CO - Histo Tech 7. Oregon - Histo Tech 8. Upstate NY - 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 do not have an email address, if you could pass my query & name on to them I'd be very grateful. Thank You !! Pam - 800 466 9919 ext 234 --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing Learn More About Ategra: Ategra Systems Inc Specialists in Permanent & Contract Staffing VOICE: 407-671-5800 ext 234 TOLLFREE: 800-466-9919 ext 234 EMAIL: pam@ategra.com WEBSITE: --------------------------------------------------------- From grantgd <@t> comcast.net Fri Oct 17 14:24:19 2003 From: grantgd <@t> comcast.net (Gordon Grant) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] LDH Staining Message-ID: <000d01c394e4$44e6da20$0300a8c0@ggrant> Does anyone have a working protocol for direct LDH staining on fresh tissue then formalin fixing and cutting. I have been attempting to get this stain protocol to work but it is not working. Gordon Grant -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031017/cfcb03ac/attachment.htm From Gillian.Barlow <@t> cshs.org Fri Oct 17 16:38:13 2003 From: Gillian.Barlow <@t> cshs.org (Barlow, Gillian) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Fresh vs. frozen paraformaldehyde Message-ID: <3CFAA0108952D111A5BF00805FA6FB0F0464D4AE@PEDSNTAS.csmc.edu> Dear Histonetters Please solve an ongoing debate for our perfusions! I was always told that 4% paraformaldehyde should be made fresh each time before using, at max made the evening before and stored at 4 degrees to use the following morning. However, somebody from another lab says they make theirs in bulk and freeze what they dont use, thawing and refreezing as necessary. Is this ok to do? They also say they use 10% neutral buffered formalin for their perfusions if they dont have 4% PFA. Many thanks Gillian Gillian 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 -------------- next part -------------- A non-text attachment was scrubbed... Name: not available Type: application/ms-tnef Size: 1982 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031017/b500fe93/attachment.bin From mrl0627 <@t> mail.ecu.edu Fri Oct 17 17:05:11 2003 From: mrl0627 <@t> mail.ecu.edu (mrl0627@mail.ecu.edu) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] RE: Fresh vs. frozen paraformaldehyde Message-ID: Hello, Gillian! My understanding is that repeated freeze-thaw hastens oxidation (per Bridget Hogan and several other texts). I freeze freshly-made 4% para in aliquots so that I thaw only once. Of course, since it has to be kept in glass, it takes up a bit of space in the freezer! Otherwise, our lab keeps fresh-made at 4 degrees for no longer than one week. No problems in last 2 years. Hope this helps. Maureen Loomer, MS candidate in Cell Biology, East Carolina U. -----Original Message----- From: "Barlow, Gillian" <Gillian.Barlow@cshs.org> To: 'Histonet' <histonet@lists.utsouthwestern.edu> Sent: Fri, 17 Oct 2003 17:38:13 -0400 Subject: Fresh vs. frozen paraformaldehyde Dear Histonetters Please solve an ongoing debate for our perfusions! I was always told that 4% paraformaldehyde should be made fresh each time before using, at max made the evening before and stored at 4 degrees to use the following morning. However, somebody from another lab says they make theirs in bulk and freeze what they dont use, thawing and refreezing as necessary. Is this ok to do? They also say they use 10% neutral buffered formalin for their perfusions if they dont have 4% PFA. Many thanks Gillian Gillian 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 theresajoseph <@t> yahoo.com Sun Oct 19 15:46:17 2003 From: theresajoseph <@t> yahoo.com (theresa joseph) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Sodium per iodate or hydrogen peroxide ? Message-ID: <20031019204617.19729.qmail@web12506.mail.yahoo.com> Hi! I have used 2.1% sodium per iodate(w/v) to quench the endogenous peroxide in the hippocampal tissues. However inthe muslce protocol, it has been suggested that I use 0.6% Hydrogen peroxide (v/v). Both sodium periodate and hydrogen peroxide seem to have similar functions. Hence I would like to know the situations where Sodiumperiodate needs to be selected instead of Hydrogen peroxide and vice versa. Thanks, Theresa J. Kannanayakal Ohio State University Columbus,OH 43210 --------------------------------- Do you Yahoo!? The New Yahoo! Shopping - with improved product search -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031019/4d1c1ab5/attachment.htm From doscwk <@t> nus.edu.sg Sun Oct 19 20:09:51 2003 From: doscwk <@t> nus.edu.sg (Chan Wai Kam) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Auto stainer and coverslipper Message-ID: Hi Histonetters, Thanks for your response to my query regarding the use of manual versus auto coverslipper. The reason I asked is that I was wondering whether an auto machine could perform as well as a human when it comes to eliminating certain unwanted stuff such as this problem of air bubbles during coverslipping. I find that the consistency of the mounting media plays a part as one that is too thick tends to give rise to air bubbles and so is how one lowers the coverslip onto the slide. Usually it's not a problem for me as I would either gently apply some force over the coverslip and push out the air bubbles, or if I'm unable to get rid of some of them, I just soak the slide in xylene and reapply another coverslip. I was just wondering whether we should get an automated machine to make things easier, that is if the machine is great to have in a histo lab. Thanks Julee Chan Orthopaedic Surgery National University of Singapore From AnthonyH <@t> chw.edu.au Mon Oct 20 00:10:46 2003 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Job Opening (Sydney):TRC Coordinator (Research Assistant / Office r) Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E06B@simba.kids> <<...OLE_Obj...>> TRC Coordinator (Research Assistant / Officer) NSW Tissue Resource Centre Department of Pathology, University of Sydney The Neuroscience Institute of Schizophrenia and Allied Disorders and the University of Sydney are coordinating human brain banking to facilitate and support neuroscience studies of neuropsychiatric disorders throughout Australia and overseas. This position is responsible for the coordination of the NSW TRC functions. This includes tissue dissection and handling (fixed and frozen tissue), liaising with research groups, and the preparation of samples for distribution. Responsibilities also include maintaining the TRC database and the implementation of quality control protocols. There will be opportunities to be involved in research initiatives of relevance to schizophrenia in the very active Neuropathology Department. Essential criteria include: histotechnology skills including management of laboratory equipment; the ability to work in a collaborative research team; excellent communication skills and good organisational skills. Desirable criteria: Neuroscience background, neuroanatomy, molecular biology techniques. For further information please contact Prof. Clive Harper (cliveh@med.usyd,edu.au) or 02-9351 3663). Salaries are in accordance with NISAD salary scales. The closing date for applications is 30 October 2003. A job description can be found on the NISAD web site (www.nisad.org.au/careers/default.asp ) Applications to be submitted to: Daren Draganic Research Manager NISAD, 384 Victoria St, Darlinghurst, NSW, 2010. Phone: (02) 9295 8408; Fax: (02) 9295 8415; E-mail: d.draganic@nisad.org.au ; Web: www.nisad.org.au Regards, Tony Henwood 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 ian.montgomery <@t> bio.gla.ac.uk Mon Oct 20 04:47:20 2003 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Eosinophils. Message-ID: <5.2.1.1.2.20031020104312.00a5e300@udcf.gla.ac.uk> Has anyone been using an eosinophil MBP successfully on fixed paraffin sections? My search, so far, for an antibody has been unsuccessful. 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 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031020/df3c865b/attachment.htm From victoria.hahn-stromberg <@t> orebroll.se Mon Oct 20 06:20:39 2003 From: victoria.hahn-stromberg <@t> orebroll.se (victoria.hahn-stromberg@orebroll.se) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] LMD Message-ID: <0875915A604AF444AAB9AB43FD6DE8C72AFEDF@storm.orebroll.se> Hi! I?m having problems with my LMD slides. I use 10um paraffin embedded histological sections which I would like to stain immunohistochemically with CAM, but the sections keep falling off, can anyone plesse help me??/Victoria Victoria Hahn-Str?mberg MSc, PhD student Kliniken f?r Patologi Universitetssjukhuset ?rebro 701 85 ?rebro tel: 019-6023644 faxnr:019-6021035 From margaret_b_wilhelms <@t> groton.pfizer.com Mon Oct 20 07:58:50 2003 From: margaret_b_wilhelms <@t> groton.pfizer.com (Wilhelms, Margaret B) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] EDTA Message-ID: I've seen lots of posts in the archives about EDTA and formulas. But I have a question: Has anyone had any trouble getting EDTA powder to dissolve in distilled water? Meg 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 Barry.R.Rittman <@t> uth.tmc.edu Mon Oct 20 08:22:21 2003 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] EDTA Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0635872@UTHEVS3.mail.uthouston.edu> Margaret EDTA itself is only slightly soluble in water. As you start to make the solution more alkaline e.g. by slowly adding sodium hydroxide solution, it will start to dissolve. Then adjust pH to what you finally want. An easier approach is to start with disodium EDTA which is very soluble in water and then adjust the pH. Barry Barry. R. Rittman 6516 MD Anderson Blvd. Houston, TX. 77030 713-500-4134 telephone 713-500-4500 fax -----Original Message----- From: Wilhelms, Margaret B [mailto:margaret_b_wilhelms@groton.pfizer.com] Sent: Monday, October 20, 2003 7:59 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] EDTA I've seen lots of posts in the archives about EDTA and formulas. But I have a question: Has anyone had any trouble getting EDTA powder to dissolve in distilled water? Meg LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TJJ <@t> Stowers-Institute.org Mon Oct 20 13:54:04 2003 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Squash preparation of germinal tissues Message-ID: Does anybody have a protocol for making squash preparations of lizard (or other reptile) ovaries for germ cell examination by Feulgen stain? Thanks for any help I get in this matter! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From histo20 <@t> hotmail.com Mon Oct 20 14:19:34 2003 From: histo20 <@t> hotmail.com (Paula Wilder) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Billing for procedures Message-ID: I don't know if anyone can help with this. We are being asked to calculate all expenses associated with every procedure done in pathology. This is to include reagents, tech time; like tests are to be grouped together. This is so that we capture all the billing charges. We have not been given any guidance or any particular place to start. Though other hospitals have develped their own activity master, we were asked to re-invent the wheel. Any help, any direction with this would be very greatly appreciated! Paula Wilder St. Joseph Medical Center Towson, MD _________________________________________________________________ Add MSN 8 Internet Software to your current Internet access and enjoy patented spam control and more. Get two months FREE! http://join.msn.com/?page=dept/byoa From amosbrooks <@t> earthlink.net Mon Oct 20 15:20:39 2003 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] LMD ... IHC sections falling off In-Reply-To: <20031020170001.19912.209.Mailman@swlx167.swmed.edu> References: <20031020170001.19912.209.Mailman@swlx167.swmed.edu> Message-ID: <6.0.0.22.0.20031020161503.029a16f0@127.0.0.1> Victoria, Firstly try thinner sections 10 um is a bit thick. Next you should use some type of charged, silanized or APES slides. You can either silanize your own slides or buy them from a vendor. Adhesives aren't necessarily good for IHC as they can interfere with the reaction. Amos Brooks >Message: 5 >From: victoria.hahn-stromberg@orebroll.se >To: histonet@lists.utsouthwestern.edu >Date: Mon, 20 Oct 2003 13:20:39 +0200 >Subject: [Histonet] LMD > >Hi! >I=B4m having problems with my LMD slides. I use 10um paraffin embedded >histological sections which I would like to stain immunohistochemically = >with >CAM, but the sections keep falling off, can anyone plesse help = >me??/Victoria > >Victoria Hahn-Str=F6mberg >MSc, PhD student >Kliniken f=F6r Patologi >Universitetssjukhuset =D6rebro >701 85 =D6rebro >tel: 019-6023644 >faxnr:019-6021035 From usdahisto <@t> hotmail.com Mon Oct 20 17:25:27 2003 From: usdahisto <@t> hotmail.com (T. Truscott) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] RE: Immunos Message-ID: For economical purposes, only run the common IHC's and send off all the ones that you don't run every week or two. Good luck, Tom Truscott, USDA-ARS >From: "Richard Cartun" >To: , >Subject: [Histonet] RE: Immunos >Date: Fri, 10 Oct 2003 12:25:25 -0400 > >Obviously, immunohistochemical stains are not required for the vast >majority of specimens that we see in the surgical pathology laboratory, >but, if I had a completed lesion removed, I would not want it sent to >your pathologist who claims that immunohistochemical stains are not >required for diagnosis! > >Richard Cartun > > >>> "Hallada, Teri" 10/10/03 10:19AM >>> >Hi all, >I have an inquiry about immunos. We have differing opinions here at our >institution. We have one pathologist that likes to utilize immunos in >diagnosing cases and one that insists that they are not required to make >a diagnosis. We currently send all of our immunos out to be stained and >read them here upon return. We are considering trying to bring them in >house, but are concerned with the volume that we are ordering. We are a >small community hospital, but have a pretty good outpatient clientele >also. I was wondering if I could get some feedback from the group about >the importance of utilizing immunos and volumes that are being ordered. >Also, do any of you use certain immunos as part of your protocol for >specific cases (other than breast). >Thank you, >Teri Hallada BS MT CT (ASCP) >thallada@noch.org > > > > ___________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Want to check if your PC is virus-infected? Get a FREE computer virus scan online from McAfee. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From usdahisto <@t> hotmail.com Mon Oct 20 18:07:40 2003 From: usdahisto <@t> hotmail.com (T. Truscott) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] STATIC AT THE MICROTOME Message-ID: Bob, You can also try raising the humidity in your lab or around your microtome.Also try soaking your paraffin blocks in water or in water on ice. Tom Truscott USDA-ARS >From: "Barbara Stancel" >To: AliNeumann@aol.com, histonet@pathology.swmed.edu >Subject: Re: [Histonet] STATIC AT THE MICROTOME >Date: Mon, 13 Oct 2003 18:24:54 +0000 > >Dear Bob, > >Dryer sheets. Try them in your pockets or stuffed up your sleeves. Rub them >on your hands as you would toweling off after washing. I have even used >them wrapped around the block (chuck) holder of the microtome or stuffed in >the paraffin tray under the knive. They come fragranced and >non-fragranced. I have also tried the anti-static sprays sold for spraying >on clothes to reduce "creeping" skirts and pants. Works pretty good sprayed >on microtomes and/or sleeves. There have been dry winters that I have used >all of the above. > >Histologically yours, Barbara > >Barbara H. Stancel, HTL(ASCP)HT >USDA, FSIS, OPHS, Eastern Laboratory, Pathology >RRC, 950 College Station Road >Athens, Georgia > > > >>From: AliNeumann@aol.com >>To: histonet@pathology.swmed.edu >>Subject: [Histonet] STATIC AT THE MICROTOME >>Date: Fri, 10 Oct 2003 23:03:54 EDT >> >>I am learning to be a histotech and when using the manual Leitz 1512 >>microtome, I seem to have a problem with alot of static, the ribbons >>either move away >>from my fingers or they jump on and wrap around them, or they jump on to >>the >>disposable kinfe holder or the frame of the microtome. Does anyone have >>any >>suggestions they would be willing to share with me, to make my learning >>less >>irritating? >>Thank You, >>Bob Neumann > >_________________________________________________________________ >Frustrated with dial-up? Get high-speed for as low as $29.95/month >(depending on the local service providers in your area). >https://broadband.msn.com > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Surf and talk on the phone at the same time with broadband Internet access. Get high-speed for as low as $29.95/month (depending on the local service providers in your area). https://broadband.msn.com From Sellis4051 <@t> aol.com Mon Oct 20 20:47:28 2003 From: Sellis4051 <@t> aol.com (Sellis4051@aol.com) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Histology's contribution to Forensic Science Message-ID: <5f.4090d57c.2cc5ea30@aol.com> I'm not currently working in histology so that I can complete a Master's degree but have volunteered to give a presentation at the next Washington State Histology meeting in May. The tentative title of the presentation is "The role of the histotech in death investigations." I parlayed my histology training into morgue work and then death scene investigations. I am now a Diplomat with the American Board of Medicolegal Death Investigation. I know that there are very few histotechs working solely for a coroner or medical examiner. When I was a histology intern at Harborview Medical Center, they let me learn to cut on the blocks from medical examiner cases because they were less important than the clinical stuff. I would love to hear from histotechs who cut medical examiner/coroner blocks and who have some suggestions on what I should include in my talk. Any suprising findings due to routine histology? Any cases that benefited from the specialized knowledge of a histotech? Perhaps Congo Red cracked the case? Has the experience from CAP inspections helped in gaining NAME accreditation? (National Association of Medical Examiners) Any suggestions would be really appreciated, even if you don't have forensic experience to draw on. Thanks so much, Sandra Ellis -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031020/f4a25b68/attachment.htm From Linresearch <@t> aol.com Mon Oct 20 21:37:06 2003 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] eNos Ab Message-ID: <65.1b40d761.2cc5f5d2@aol.com> Hi, Any advice on eNos in FFPE rodent tissues would be appreciated. Lin -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031020/a469bbce/attachment.htm From DDDeltour <@t> sig.med.navy.mil Tue Oct 21 00:00:42 2003 From: DDDeltour <@t> sig.med.navy.mil (Deltour, Douglas D.(HM2)) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Histology's contribution to Forensic Science Message-ID: I would suggest that you give these guys a call or email. http://www.afip.org/Departments/oafme/ HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy LPO Histology Histology Technician DSN 624-4669 FAX 624-4680 COMM (From US) 011-39-095-56-4669 DISCLAIMER: 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. **** Informazione Confidenziale - Questa e-Mail, incluso eventuali allegati, contiene informazioni confidenziali intese unicamente alla persona/e a cui e' indirizzata. Se avete ricevuto per errore questo messaggio, cortesemente contattare immediatamente il mittente e cancellare la e-Mail. Grazie****. -----Original Message----- From: Sellis4051@aol.com [mailto:Sellis4051@aol.com] Sent: Tuesday, October 21, 2003 3:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histology's contribution to Forensic Science I'm not currently working in histology so that I can complete a Master's degree but have volunteered to give a presentation at the next Washington State Histology meeting in May. The tentative title of the presentation is "The role of the histotech in death investigations." I parlayed my histology training into morgue work and then death scene investigations. I am now a Diplomat with the American Board of Medicolegal Death Investigation. I know that there are very few histotechs working solely for a coroner or medical examiner. When I was a histology intern at Harborview Medical Center, they let me learn to cut on the blocks from medical examiner cases because they were less important than the clinical stuff. I would love to hear from histotechs who cut medical examiner/coroner blocks and who have some suggestions on what I should include in my talk. Any suprising findings due to routine histology? Any cases that benefited from the specialized knowledge of a histotech? Perhaps Congo Red cracked the case? Has the experience from CAP inspections helped in gaining NAME accreditation? (National Association of Medical Examiners) Any suggestions would be really appreciated, even if you don't have forensic experience to draw on. Thanks so much, Sandra Ellis -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031021/183f62a4/attachment.htm From tissuearray <@t> hotmail.com Tue Oct 21 07:27:45 2003 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Tissue Array Questions? Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031021/ba4b1ebf/attachment.htm From gudrun.lang <@t> aon.at Tue Oct 21 07:33:46 2003 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] dialysed ironsolution for Hale's Message-ID: <000c01c397cf$929f1730$0d12a8c0@SERVER> Dear histonetters! I need some advice with the Hale's stain. I have found the procedures in the archives, but unfortunately I have no idea what "dialysed" in German means. Please tell me the detailled procedure, when the text say: "This mixture is dialysed against regularly changed water for three days." Gundi Lang general hospital, Linz, Austria -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031021/c47a6792/attachment.htm From louise_renton <@t> hotmail.com Tue Oct 21 08:42:22 2003 From: louise_renton <@t> hotmail.com (louise renton) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] poor sectioning Message-ID: Hi all, I am attempting to cut various polyethylene and other non-organic tissue implants embedded in MMA (technovit 9100) using a motorised sledge microtome and tungsten carbide knife. HOWEVER, I find that I am not getting whole sections, and that the knife is "skipping" parts i.e cutting thick and thin in one section and alternating thickness between sections as well. I have tried tightening all clamps and screws as well as my abdominal muscles, all to no avail. What is wrong? Could it be: a) too great a varance in consistencies throughout the block, ie embedding material/implant hardness mismatch b) problems with polymerization c) incorrect knife angle ( I tried adjusting this) d) wrong phase of the moon Am I the only one experienceing this problem and does anyone have suggestions to help improve my sections?. BTW, I am trying to avoid sawing and grinding techniques that are way too time consuming and tedious. TIA Louise Renton Bone Research Unit MRC Johannesburg South Africa Tel & fax +27 11 717 2298 "Time flies like an arrow, fruit flies like a banana" _________________________________________________________________ Find a new partner with MSN Personals. http://www.msn.co.za/personals/ From asmith <@t> mail.barry.edu Tue Oct 21 09:44:42 2003 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] dialysed ironsolution for Hale's Message-ID: <494304423C63E246A5CF87A3AEEB577011B59B@bumail01.barrynet.barry.edu> Dialysis is the movement of solutes through a semi-permeable membrane. The objective is to eliminate particles too small to be useful in the staining reaction. To make dialyzed iron one pours the ferric hydroxide suspension into seamless cellulose tubing with 2.4 nm pore radius (i.e. 4.8 nm pore diameter) and ties off the end of the tubing. (The other end was tied off before you began to pour.) One immerses the tubing in distilled water for 3 days, changing the water 9 times a day. To make 532 ml of colloidal iron suspension you will need about a 55 cm length of 23.5 mm radius (i.e., 47 mm diameter) seamless cellulose tubing, which you can tie off 5 cm from each end. You should dialyze against about 15 litres of distilled water. After the dialysis step one filters the suspension through Whatman #1 (medium porosity filter paper) to eliminate particles that are too large to be useful. The ferric hydroxide suspension is made by dissolving 100 g ferric chloride hexahydrate in 333 ml distilled water and adding 133 ml glycerol. One then adds 4 aliquots of concentrated ammonium hydroxide: 33 ml, 17 ml, 10 ml, and 6.2 ml; stirring until the precipitate is resuspended after each addition. The staining suspension is made by adding 7 ml glacial acetic acid to 28 ml of the dialyzed stock suspension. REFERENCE: J.F. Rinehart & S.K. Abul-Haj (1951) An improved method for histologic demonstration of acidic mucopolysaccharides in tissues. Arch. Pathol. 52: 189-194. Personally, I think that dialyzing the ferric hydroxide is a lot of work for little advantage. I usually use Mowry's quick and dirty method which can be found in Humason's "Animal Tissue Techniques." The quick and dirty method works well 90% of the time. ORIGINAL REFERENCE: R.W. Mowry (1958) J.Clin.Invest. 7: 566-576. -----Original Message----- From: Gudrun Lang [mailto:gudrun.lang@aon.at] Sent: Tuesday, October 21, 2003 8:34 AM To: Histonetliste Subject: [Histonet] dialysed ironsolution for Hale's Dear histonetters! I need some advice with the Hale's stain. I have found the procedures in the archives, but unfortunately I have no idea what "dialysed" in German means. Please tell me the detailled procedure, when the text say: "This mixture is dialysed against regularly changed water for three days." Gundi Lang general hospital, Linz, Austria 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) -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031021/392a5618/attachment.htm From c.m.vanderloos <@t> amc.uva.nl Tue Oct 21 10:10:16 2003 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] RE: fixative for insects Message-ID: Hi Andrea, I was once visiting an entomologist institute in Budjeovice (yes were the Budweiser is originally coming from!) in former Tjechoslovakia. Those guys were fixing all kinds of insects (or part of insects) in Bouin-fixative and have them then processed to paraffin as usual. Good luck! Chris van der Loos Andrea Grantham wrote: > I'm going to be processing whole mosquitoes and I was wondering what > would be the best fixative to use...and processing schedule if > anybody has one. > > Thanks. > Andi > ..................................................................... > : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > : Sr. Research Specialist University of Arizona : > : (office: AHSC 4212) P.O. Box 245044 : > : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) From MGomez <@t> ameripath.com Tue Oct 21 10:59:03 2003 From: MGomez <@t> ameripath.com (MGomez@ameripath.com) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Gravity Alcohol Filter Message-ID: Good morning Histonetters: I'm looking to replace a couple of Alcohol filters that work with gravity. Do you use them or know anyone who sells them.? Thank you very much, Milton 801-256-0040x254 From cgfields <@t> lexhealth.org Tue Oct 21 12:02:48 2003 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Cryostat decontamination Message-ID: In the last few months there have been a few procedures posted on the Net for cryostat decontamination. I thought that I had saved the procedure and I may have, but I cannot put my hands on it. Would someone be so kind to re-post the procedure please. Thank you Carole Fields Lexington Med. Ctn W.Columbia, SC _________________________________ 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. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031021/328c72c8/attachment.htm From CCLYATT <@t> mail.mcg.edu Tue Oct 21 12:36:17 2003 From: CCLYATT <@t> mail.mcg.edu (Claye Clyatt) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Gravity Alcohol Filter Message-ID: I think NewComers Supply has them. 800-383-7799. Claye >>> 10/21/03 11:59AM >>> Good morning Histonetters: I'm looking to replace a couple of Alcohol filters that work with gravity. Do you use them or know anyone who sells them.? Thank you very much, Milton 801-256-0040x254 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RizoC <@t> chi.osu.edu Tue Oct 21 12:46:49 2003 From: RizoC <@t> chi.osu.edu (Rizo, Christian) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Dynamic Histotechnologist Position in Columbus, Ohio Message-ID: <3F8707A1ADC19C4FA84BC95B51CEF56007C1E4DA@chi2kms03.columbuschildrens.net> Hello Histonetters, This is my first time writing on the lists so pardon my language. But Childrens Hospital Columbus has an opening for a Histotechnician or Histotechnologist. There is a first shift (9:30 A.M. - 6:00 P.M.) position in the Anatomic Pathology Laboratory of Childrens Hospital Columbus, Ohio that is immediately available. The position is a full-time position, working Monday through Friday. No weekend. Applicants must be a HTL (ASCP) or HT (ASCP) or eligible, preference for B.S. degree. Certification or eligibility for certification as a Histotechnologist or Histotechnician by ASCP is required. Non-certified candidate will be required to pursue certification within the first year of employment. This position is responsible for varied laboratory procedures, including specimen processing, microtomy, H&E staining, special stains and immunostaining. Duties may change depending on workload. Applicants should be dynamic, creative and enthusiastic. Good customer relations, computer skills, teamwork, dependability and communication are necessary for successful candidates. All Histology technologists share holiday coverage responsibility. All qualified and interested applicants should submit a letter of interest to: Childrens Hospital Columbus 700 Childrens Drive Columbus, OH 43205 ATTN: Josette Alexander Or e-mail me at: rizoc@chi.osu.edu Columbus is a wonderful environment for living, playing, and working. As the state capital of Ohio, the city has much to offer everyone from all stages and all walks of life. Check out the links below to see what other organizations have to share about Columbus. The Official Website for the City of Columbus, Ohio This site contains information from city officials and government agencies. The "Columbus Supersite" Sponsored by the Greater Columbus Chamber of Commerce, this site contains vast resources of general information, events, and businesses in the city. The Greater Columbus Chamber of Commerce According to the site, "The Chamber leads and supports economic growth & development for the Greater Columbus Community in the global marketplace. CitySearch Search an extended listing of events, restaurants, venues, hotels, and movies; provides suggestions for the "Best of..." around the city. About Columbus As part of the larger about.com site, the section on Columbus gives an overview of the issues, topics, restaurants, and entertainment venues that are forefront in Central Ohio. Columbus/Franklin County News Bureau Designed for journalists, this site offers reliable statistics and information about Columbus and the surrounding Franklin County. Columbus, Ohio, USA This site may help you to discover Columbus and central Ohio with its information, links, community guide, and resource center. ColumbusPage This site has a searchable and fairly complete listing of entertainment around the city, as well as links to important city services. Christian M. Rizo MBA, HTL (ASCP) Manager, Anatomic Pathology Childrens Hospital 700 Childrens Drive Columbus, Ohio 43205 614.722.5465 RizoC@chi.osu.edu Confidentiality Notice: This e-mail message, from Children's Hospital, Columbus, Ohio, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. The recipient is responsible to maintain the confidentiality of this information and to use the information only for authorized purposes pursuant to Children's Hospital's confidentiality policies. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that any review, use, disclosure, distribution, copying, printing, or action taken in reliance on the contents of this e-mail is strictly prohibited. If you have received this communication in error, please notify us immediately by reply e-mail and destroy all copies of the original message. Thank you. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031021/8a44df0e/attachment.htm From Lynne.Bell <@t> hitchcock.org Tue Oct 21 12:55:44 2003 From: Lynne.Bell <@t> hitchcock.org (Bell, Lynne) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Gravity Alcohol Filter Message-ID: Newcomer Supply has them - I just received a new catalog today. The website is www.newcomersupply.com, phone number is 1-800-383-7799. Nicest bunch of people I have ever had the pleasure to do business with!! Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 From cfavara <@t> niaid.nih.gov Tue Oct 21 12:36:56 2003 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] non-human primates Message-ID: I am trying to collect information on the processing and staining of non-human primate brain. The proposal is to look at half-coronal sections of macaque and squirrel monkeys. I am more concerned with the size of mold and slides that will be necessary. I seem to recall a company that specializes in nueropathology but cannot seem to locate it now. Any information would be greatly appreciated! Thanks, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 From alust1 <@t> hotmail.com Tue Oct 21 12:11:15 2003 From: alust1 <@t> hotmail.com (Andrew Lust) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] exposing epitope region on MCL of rat Message-ID: Dear Histonetters I have a quick question, I was wondering if anybody has worked with Medial collateral ligament of the rat in immuno-fluoresces staining? If you have, did you have to do any type of epitope exposure to the tissue? I know companies offer pre made solutions, but have no clue if I need to use this solution. Considering that I am not going there paraffin embedded tissue, so I am not exposing the tissue to harsh conditions that would degrade proteins on my tissue. Staining so far has been questionable, so I am up for any suggestions on fluoresce staining on the MCL of the Rat. thanks again Andrew L lab tech University of Houston _________________________________________________________________ Never get a busy signal because you are always connected with high-speed Internet access. Click here to comparison-shop providers. https://broadband.msn.com From cwscouten <@t> myneurolab.com Tue Oct 21 13:33:13 2003 From: cwscouten <@t> myneurolab.com (Charles W. Scouten, Ph.D.) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Cryostat decontamination Message-ID: Why should you decontaminate your cryostat? Let the durn thing do the work itself while you sleep. See the attached link. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=4 75102&catdesc=Histology+Equipment&CatThreeID=609&CatOneID=4&subcatdesc=C ryostats&idsubcategory=182 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 www.myneurolab.com -----Original Message----- From: Carole Fields [mailto:cgfields@lexhealth.org] Sent: Tuesday, October 21, 2003 12:03 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cryostat decontamination In the last few months there have been a few procedures posted on the Net for cryostat decontamination. I thought that I had saved the procedure and I may have, but I cannot put my hands on it. Would someone be so kind to re-post the procedure please. Thank you Carole Fields Lexington Med. Ctn W.Columbia, SC _________________________________ 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. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031021/b3549128/attachment.htm From andrae <@t> u.washington.edu Tue Oct 21 15:01:28 2003 From: andrae <@t> u.washington.edu (A. Erickson) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] non-human primates In-Reply-To: References: Message-ID: Brain Research Labs Newton, Ma 02468 (617)965-5544 has a good catalog. I also got some huge "Macroflow' cassettes from Microm via Richard allen. My son (a machinist) made some plates & molds for me. Regards, andra erickson, WaNPRC UW, Seattle, Wa. On Tue, 21 Oct 2003, Favara, Cynthia (NIH/NIAID) wrote: > I am trying to collect information on the processing and staining of > non-human primate brain. The proposal is to look at half-coronal sections of > macaque and squirrel monkeys. I am more concerned with the size of mold and > slides that will be necessary. > > I seem to recall a company that specializes in nueropathology but cannot > seem to locate it now. > > Any information would be greatly appreciated! > > Thanks, > c > > Cynthia Favara > NIAID/NIH/RML/LPVD > 903 South 4th Street > Hamilton, MT 59840 > 406-363-9317 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From sa.drew <@t> hosp.wisc.edu Tue Oct 21 15:39:12 2003 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Anyone have a favorite CD15 clone? Message-ID: We are just wondering if not all clones of CD15 are equal, and that maybe someone out there has tested more than a couple different CD15's? We have pathologists who periodically ask us to repeat our stain because they expect to see more RS cells staining than they do. We currently run our antibodies on Ventana immunostainers if anyone feels that makes a difference. All constructive opinions are welcome. Thank you! Sally Ann Drew-MT(ASCP) Univ. of Wisconsin Hospital & Clinics IHC/ISH Laboratory 600 Highland Ave. VAH-DM223 Madison, WI 53792-2472 608-265-6596 sa.drew@hosp.wisc.edu From amarusk1 <@t> FAIRVIEW.ORG Tue Oct 21 17:14:33 2003 From: amarusk1 <@t> FAIRVIEW.ORG (ANN MARUSKA) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] MOC-31 Message-ID: Skipped content of type multipart/alternative-------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:ANN MARUSKA EMAIL;WORK;PREF;NGW:AMARUSK1.EAST.NORTH ORG:;LAB N:MARUSKA;ANN TEL;WORK:612-273-9119 END:VCARD From emry <@t> u.washington.edu Tue Oct 21 17:36:11 2003 From: emry <@t> u.washington.edu (P. Emry) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Brdu Message-ID: I have been told that there are problems using our formaldehyde substitute with BrDu staining. I hate going back to the evil stuff. Is anyone doing BrDu with a substitute that works? Thanks, Trisha Seattle From maijazi <@t> hotmail.com Tue Oct 21 20:19:32 2003 From: maijazi <@t> hotmail.com (Mahmood Aijazi) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Immunohistochemistry Home Page Message-ID: Please check out our new home of Immunohistochemistry Home Page at http://www.immunocentral.com We have installed a powerful discussion forum with lots of subcategories. We had to close our previous site abruptly due to problems with our hosting company. Please spread the word so that more and more people can get the benefit of this site. Mahmood Aijazi, MD _________________________________________________________________ See when your friends are online with MSN Messenger 6.0. Download it now FREE! http://msnmessenger-download.com From Gillian.Barlow <@t> cshs.org Tue Oct 21 21:12:14 2003 From: Gillian.Barlow <@t> cshs.org (Barlow, Gillian) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Problems with immunohistochemistry on formalin-fixed heart sectio ns Message-ID: <3CFAA0108952D111A5BF00805FA6FB0F0464D4B7@PEDSNTAS.csmc.edu> Dear Histonetters This is the latest installment in our ongoing heart saga, I hope someone out there can help!!! Following advice from you all (many thanks!) we now follow the following protocol: we harvest our hearts from live but anaesthetized animals so that they are still beating, cut them in half to allow the fixative access to the tissues, and fix in 10% buffered formalin for usually 12 to 24 hours at room temp. After this, we process, embed in paraffin and section at 8 microns. The preservation of the tissue looks good when we stain with H&E, but when we do the immunohistochemistry, the cells look destroyed. Commercial sections purchased from Novagen, who fix their hearts in 4% paraformaldehyde, look fine using identical immunohistochemical techniques. Are formalin-fixed hearts known to be more sensitive e.g. to pretreatment with peroxide to block endogenous peroxidases? Or is there some other step likely to be causing trouble? The antibodies are from Santa Cruz, and are supposed to work well on formalin- or PFA-fixed tissues. All suggestions gratefully received! Many thanks Gillian Gillian 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 -------------- next part -------------- A non-text attachment was scrubbed... Name: not available Type: application/ms-tnef Size: 2524 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031021/0c281309/attachment.bin From AnthonyH <@t> chw.edu.au Tue Oct 21 22:56:20 2003 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Anyone have a favorite CD15 clone? Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E079@simba.kids> Sally, We compared two CD15 clones on several Hodgkin's disease cases and found LeuM1 the best. Staining overall was improved with both antibodies if overnight incubation (4oC) was used. Remember that most CD15 antibodies are IgM (not IgG), and therefore bigger. Also the detection system may not be all that efficient for IgM antibodies. 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: Drew Sally A. [mailto:sa.drew@hosp.wisc.edu] Sent: Wednesday, 22 October 2003 6:39 To: Histonet Subject: [Histonet] Anyone have a favorite CD15 clone? We are just wondering if not all clones of CD15 are equal, and that maybe someone out there has tested more than a couple different CD15's? We have pathologists who periodically ask us to repeat our stain because they expect to see more RS cells staining than they do. We currently run our antibodies on Ventana immunostainers if anyone feels that makes a difference. All constructive opinions are welcome. Thank you! Sally Ann Drew-MT(ASCP) Univ. of Wisconsin Hospital & Clinics IHC/ISH Laboratory 600 Highland Ave. VAH-DM223 Madison, WI 53792-2472 608-265-6596 sa.drew@hosp.wisc.edu _______________________________________________ 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 i_stain <@t> yahoo.com Wed Oct 22 02:30:52 2003 From: i_stain <@t> yahoo.com (Scott Mordue) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Cyclin D1 Message-ID: <20031022073052.84332.qmail@web42004.mail.yahoo.com> Hi Mary, We get our Cyclin D1, clone AM29 from Zymed. Try order it from them. Scott CSU -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Mary Bryhan Sent: Thursday, October 16, 2003 12:22 PM To: r.min@biogenex.com Cc: Judith Hoschner; Marcia Shattuck; Adolfo Noel Ceniza MD; histonet@pathology.swmed.edu Subject: [Histonet] Cyclin D1 Dear Roy, I found out today through our purchasing department that Cyclin D1 has been discontinued by BioGenex. I called and spoke with Amanda in technical support who said that the antibody had problems and was pulled. This is not the first time that an antibody, or kit constituent has been identified as "bad" without any notice to us, the customer. If we are to keep our working relationship, I expect that BioGenex will notify us as soon as a problem is detected. Cordially, Mary Bryhan __________________________________ Do you Yahoo!? The New Yahoo! Shopping - with improved product search http://shopping.yahoo.com From abright <@t> brightinstruments.com Wed Oct 22 04:18:27 2003 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] poor sectioning Message-ID: Dear Louise & Histonetters that are interested, I have a wide experience of sectioning plastics and non-organic materials, we manufacture a range of cryostats and microtomes to suit this type of application and have supplied them for plastic implant studies too. I understand the problems you are having and to be sure to give you the correct advice I would like you to send me samples of the materials you are having problems with, please include instructions as to what face to section and how thick. Best Regards Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: louise renton [mailto:louise_renton@hotmail.com] Sent: 21 October 2003 14:42 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] poor sectioning Hi all, I am attempting to cut various polyethylene and other non-organic tissue implants embedded in MMA (technovit 9100) using a motorised sledge microtome and tungsten carbide knife. HOWEVER, I find that I am not getting whole sections, and that the knife is "skipping" parts i.e cutting thick and thin in one section and alternating thickness between sections as well. I have tried tightening all clamps and screws as well as my abdominal muscles, all to no avail. What is wrong? Could it be: a) too great a varance in consistencies throughout the block, ie embedding material/implant hardness mismatch b) problems with polymerization c) incorrect knife angle ( I tried adjusting this) d) wrong phase of the moon Am I the only one experienceing this problem and does anyone have suggestions to help improve my sections?. BTW, I am trying to avoid sawing and grinding techniques that are way too time consuming and tedious. TIA Louise Renton Bone Research Unit MRC Johannesburg South Africa Tel & fax +27 11 717 2298 "Time flies like an arrow, fruit flies like a banana" _________________________________________________________________ Find a new partner with MSN Personals. http://www.msn.co.za/personals/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ian.montgomery <@t> bio.gla.ac.uk Wed Oct 22 06:25:48 2003 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:22:04 2005 Subject: Fwd: [Histonet] Problems with immunohistochemistry on formalin-fixed heart sectio ns Message-ID: <5.2.1.1.2.20031022110948.00a6e2d0@udcf.gla.ac.uk> Skipped content of type multipart/alternative-------------- next part -------------- A non-text attachment was scrubbed... Name: [Histonet] Problems with immuno Type: application/octet-stream Size: 2524 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031022/cfcfff0f/HistonetProblemswithimmuno.obj From pam <@t> ategra.com Wed Oct 22 08:57:47 2003 From: pam <@t> ategra.com (Pam Barker (extension 234)) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Histology openings Latest Update 10/22/03 Message-ID: Hi histonetters, I am presently on a search for 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. These 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 supervisory, team lead and bench positions. These positions require HTL or HT certification or registry eligibility. Here are my NEWEST openings: 1. Las Vegas, NV - Histo Tech 2. Minneapolis, MN - Histo Tech Here are some of my hottest Histology Supevisory positions: 1. Maine - Histology Supervisor 2. Syracuse, NY - Histology Supervisor 3. Northern NJ - Histology Supervisor or Team Lead 4. Atlanta, GA - Team Lead grossing required 5. New Hampshire - Histo Supervisor 6. Las Vegas, NV - Histo Supervisor Here are some of my hottest Histo Tech bench positions: 1. Southwest FL - opportunity to obtain QIHC some grossing required 2. Pittsburgh, PA - Histo Tech 3. Richmond, Va - Lead Histo Tech 4. Salt Lake City, UT - Histo Tech 5. Illinois - Team Lead 6. Boulder, CO - Histo Tech 7. Oregon - Histo Tech 8. Upstate NY - Histo Tech 9. NYC, NY - 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 to pam@ategra.com (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 do not have an email address, if you could pass my query & name on to them I'd be very grateful. Thank You !! Pam - 800 466 9919 ext 234 --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing Learn More About Ategra: Ategra Systems Inc Specialists in Permanent & Contract Staffing VOICE: 407-671-5800 ext 234 TOLLFREE: 800-466-9919 ext 234 EMAIL: pam@ategra.com WEBSITE: --------------------------------------------------------- From Ronnie_Houston <@t> bshsi.com Wed Oct 22 10:56:24 2003 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] MSB dyes Message-ID: <530361BF03351B4CAE5270A05D3037B5FE04BD@bsrexms01.BSHSIR.COM> Hi Folks, I know a lot of you will be in Kentucky just now, but I need your assistance. I'm looking for a source, in the USA, of the following dyes for a MSB stain for fibrin. So far I haven't had much luck, only locating Brilliant crystal scarlet: Martius yellow (acid yellow 24) CI 10315 Brilliant crystal scarlet (acid red 44) CI 16250 Methyl blue (acid blue 93) CI 42780 Thanks Ronnie Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 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 REEVEML <@t> shands.ufl.edu Wed Oct 22 12:38:12 2003 From: REEVEML <@t> shands.ufl.edu (Mary Reeves) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] PPE Requirements in the Histology Lab Message-ID: I would like to know what types of PPEs are required in other labs. For example, while doing special stains we require the tech to wear a lab coat or scrubs and nitrile gloves. We have written PPE requirements for each task in the histo lab and I am being questioned on wether the requirements are enough. So what is everyone else requiring and how do you enforce the policy? Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 From GDawson <@t> Milw.Dynacare.com Wed Oct 22 14:14:02 2003 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] CD15 Clone Message-ID: Sally, My favorite CD15 clone is Leu M1 from Becton Dickenson (Cat. #347420). Glen Dawson From slamont <@t> u.washington.edu Wed Oct 22 15:35:33 2003 From: slamont <@t> u.washington.edu (Sarah E Lamont) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] pig neutrophil labeling Message-ID: Has anyone had experience labeling neutrophils in pig skin? I would like to use an immunohistochemistry method to quantify the number of neutrophils in the skin. I'm using frozen sections, and have tried using rabbit anti-human myeloperoxidase from Dako on PFA fixed and unfixed tissue with little success. Thanks in advance for any advice you could share! Sarah Lamont Research Scientist Department of Bioengineering University of Washington Seattle, WA From amarusk1 <@t> FAIRVIEW.ORG Wed Oct 22 15:43:41 2003 From: amarusk1 <@t> FAIRVIEW.ORG (ANN MARUSKA) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Wilms' Tumor WT1 Message-ID: Skipped content of type multipart/alternative-------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:ANN MARUSKA EMAIL;WORK;PREF;NGW:AMARUSK1.EAST.NORTH ORG:;LAB N:MARUSKA;ANN TEL;WORK:612-273-9119 END:VCARD From rschoon <@t> email.unc.edu Wed Oct 22 15:43:52 2003 From: rschoon <@t> email.unc.edu (rschoon@email.unc.edu) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Brdu In-Reply-To: References: Message-ID: <1066855432.3f96ec08afd07@webmail8.isis.unc.edu> Trisha, Which substitute are you using? I have found that BrdU is a very hardy (and therefore easly demonstrated) epitope. I have done BrdU IHC on tissues which were fixed in several different fixatives as well as on frozen sections and not had any problems. If you would identify the fixative yuo may find that someone on the list has already done this and may be able to save you some development time. Robert Schoonhoven Quoting "P. Emry" : > I have been told that there are problems using our formaldehyde > substitute > with BrDu staining. I hate going back to the evil stuff. > Is anyone doing BrDu with a substitute that works? > > Thanks, > > Trisha > Seattle > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From nmuvarak <@t> facstaff.wisc.edu Wed Oct 22 16:16:04 2003 From: nmuvarak <@t> facstaff.wisc.edu (NIDAL E MUVARAK) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] MMP-2 and MMP-9 on mouse tissue Message-ID: <8ae078743c.8743c8ae07@wiscmail.wisc.edu> I was wondering if anyone out there is working on detecting MMP-2 and -9 in mouse tissue immunohistochemically. I would be very grateful if you can let me know. I have a couple of questions regarding the procedure. Thank you for your time. Regards, 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 asmith <@t> mail.barry.edu Wed Oct 22 16:35:56 2003 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] MSB dyes Message-ID: <494304423C63E246A5CF87A3AEEB577011B59D@bumail01.barrynet.barry.edu> Martius yellow is cat. # 37,776-7 @ $36 for 25g from Aldrich (1-800-558-3850). Crystal scarlet is cat. # 21,016-1 @ $32.20 for 5g from Aldrich. Methyl blue is cat. # M6900 @ $30.60 for 50g from Sigma (1-800-325-3010). -----Original Message----- From: Houston, Ronnie [mailto:Ronnie_Houston@bshsi.com] Sent: Wednesday, October 22, 2003 11:56 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] MSB dyes Importance: High Hi Folks, I know a lot of you will be in Kentucky just now, but I need your assistance. I'm looking for a source, in the USA, of the following dyes for a MSB stain for fibrin. So far I haven't had much luck, only locating Brilliant crystal scarlet: Martius yellow (acid yellow 24) CI 10315 Brilliant crystal scarlet (acid red 44) CI 16250 Methyl blue (acid blue 93) CI 42780 Thanks Ronnie Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 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. _______________________________________________ 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 r.lederer <@t> uq.edu.au Wed Oct 22 19:06:20 2003 From: r.lederer <@t> uq.edu.au (Rose Lederer) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] amyloid staining In-Reply-To: <20031022170001.2312.24453.Mailman@swlx167.swmed.edu> Message-ID: <000001c398f9$7ce99f70$89a6a8c0@vet.uq.edu.au> Dear list, I'm new to the list, am working on a PhD in Vet science, and currently looking at the location and amount of amyloid deposits in cat pancreas (diabetic and non-diabetic) and wanted to find out what you would recommend. I know Congo red (CR) being the recommended staining method, but I didn't get good birefringence under polarised light, and the red isn't really bright so that it would be harder to evaluate it using computer imaging software. I was told to try out Sirius Red am still waiting for the package to arrive. I stained some sections with Sodium Sulfate Alcian Blue (AB) and got beautiful results, however: how unspecific is it? I'd love to be able to use it instead of Congo Red! It seems to me that the areas stained with Congo Red were exactly the ones staining blue in Alcian Blue..... I will be comparing 10 sections of CR versus AB today....and We are going to try out amylin antibody staining today! Your advice is highly welcome !!! Thanks! Rose Rose Lederer, Dr. med. vet., PhD cand. Diabetes and Obesity Group Centre for Companion Animal Health School of Veterinary Science The University of Queensland St. Lucia, Qld, 4072 Australia Ph: +61 (0) 7- 3346 9606 Fax: +61(0) 7 3365 1255 Mob: 0422 28 7371 From AnthonyH <@t> chw.edu.au Wed Oct 22 20:05:11 2003 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] amyloid staining Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E07E@simba.kids> Rose, I have found the SAB stain to be excellent for senile plaques in brain. I have not seen any study comparing the efficiency of the SAB with the CR. I would love to see your results published and would fill a hole in our knowledge. 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: Rose Lederer [mailto:r.lederer@uq.edu.au] Sent: Thursday, 23 October 2003 10:06 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] amyloid staining Dear list, I'm new to the list, am working on a PhD in Vet science, and currently looking at the location and amount of amyloid deposits in cat pancreas (diabetic and non-diabetic) and wanted to find out what you would recommend. I know Congo red (CR) being the recommended staining method, but I didn't get good birefringence under polarised light, and the red isn't really bright so that it would be harder to evaluate it using computer imaging software. I was told to try out Sirius Red am still waiting for the package to arrive. I stained some sections with Sodium Sulfate Alcian Blue (AB) and got beautiful results, however: how unspecific is it? I'd love to be able to use it instead of Congo Red! It seems to me that the areas stained with Congo Red were exactly the ones staining blue in Alcian Blue..... I will be comparing 10 sections of CR versus AB today....and We are going to try out amylin antibody staining today! Your advice is highly welcome !!! Thanks! Rose Rose Lederer, Dr. med. vet., PhD cand. Diabetes and Obesity Group Centre for Companion Animal Health School of Veterinary Science The University of Queensland St. Lucia, Qld, 4072 Australia Ph: +61 (0) 7- 3346 9606 Fax: +61(0) 7 3365 1255 Mob: 0422 28 7371 _______________________________________________ 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 katri <@t> cogeco.ca Wed Oct 22 20:17:40 2003 From: katri <@t> cogeco.ca ( Katri Tuomala) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Wilms' Tumor WT1 References: Message-ID: <008101c39903$7567e2c0$ce989618@hala4.on.cogeco.ca> Hi Ann, We use normal human kidney, where the cell nuclei in the glomeruli are positive for this antibody. Katri Katri Tuomala St.Joseph's Healthcare Hamilton, Ontario, Canada ----- Original Message ----- From: "ANN MARUSKA" To: Sent: Wednesday, October 22, 2003 4:43 PM Subject: [Histonet] Wilms' Tumor WT1 > Hi Histonetters, > > I am working up Wilms' Tumor (WT1) - and wondering what other labs are > using for control. Unfortunately, a Wilms tumor is not an option for > me. Anyone using a mesothelioma or some other tissue for a control with > this antibody? > Thanks in advance. > > Ann > > Ann Maruska > Fairview-University Medical Center > Mpls. MN 55454 > amarusk1@fairview.org > 612-273-9119 > From i_stain <@t> yahoo.com Wed Oct 22 21:13:28 2003 From: i_stain <@t> yahoo.com (Scott Mordue) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] MOC-31 Message-ID: <20031023021328.88543.qmail@web42006.mail.yahoo.com> Hi Ann, We use MOC-31 (Zymed 18-0270) 1:40 dilution P1 8 min, and using 32 min protocol. Scott Date: Tue, 21 Oct 2003 17:14:33 -0500 From: "ANN MARUSKA" To: Subject: [Histonet] MOC-31 This is a MIME message. If you are reading this text, you may want to consider changing to a mail reader or gateway that understands how to properly handle MIME multipart messages. --=_ACF20BC1.ACCDA2B3 Content-Type: multipart/alternative; boundary="=_ACF20BC1.ABCAA5B4" --=_ACF20BC1.ABCAA5B4 Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Hi Histonetters, Anyone out there (who isn't in Kentucky at the NSH) using Ventana stainers for human Epithelial Related Antigen (MOC-31) and could share some info - I would greatly appreciate. Thanks. Ann Ann Maruska Fairview-University Medical Center Mpls. MN 55454 amarusk1@fairview.org 612-273-9119 __________________________________ Do you Yahoo!? The New Yahoo! Shopping - with improved product search http://shopping.yahoo.com From d.catmull <@t> latrobe.edu.au Wed Oct 22 23:36:12 2003 From: d.catmull <@t> latrobe.edu.au (Deanne Catmull) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Bielschowsky stain Message-ID: <3.0.6.32.20031023143612.0098fce8@pop.latrobe.edu.au> Hi everyone, Was just wondering if anyone has heard of or knows of a method of bielschowsky staining that can be used on 1 micron resin-embedded sections??? Thanks, Deanne. Deanne Catmull Neuroimmunology Laboratory Dept of Biochemistry La Trobe University Bundoora Vic 3086 Ph:9479-1155 From arme <@t> optonline.net Thu Oct 23 01:42:07 2003 From: arme <@t> optonline.net (Mark Sofferman) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] For sale: VIP 3000 Tissue Processor In-Reply-To: <20031020170001.19912.209.Mailman@swlx167.swmed.edu> Message-ID: Table Top unit. Best offer. 201-833-1550 ARME Mark S. From Jackie.O'Connor <@t> abbott.com Thu Oct 23 06:46:30 2003 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] Brdu Message-ID: I'm actually doing BrDu with a zinc based fixative (that I hate) but staining is good. Contact me and I'll forward you the particulars. Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics 847.938.4919 Fax 847.938.3266 "P. Emry" Sent by: histonet-admin@lists.utsouthwestern.edu 10/21/2003 05:36 PM To: HistoNet@Pathology.swmed.edu cc: Subject: [Histonet] Brdu I have been told that there are problems using our formaldehyde substitute with BrDu staining. I hate going back to the evil stuff. Is anyone doing BrDu with a substitute that works? Thanks, Trisha Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031023/8caaf9c2/attachment.htm From Stanley.Lupo <@t> gsk.com Thu Oct 23 08:48:23 2003 From: Stanley.Lupo <@t> gsk.com (Stanley.Lupo@gsk.com) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] MSB dyes Message-ID: Stanley.Lupo@GSK.Com Safety Assessment Mail Code: UE0461 Phone: 610 270-7340 Fax: 610 270-7202 Contact Dick Dapson of Anatech Ltd. dickdapson@anatechltdusa.com 800-262-8324 I'm sure he can direct you in your search. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031023/177ff23f/attachment.htm From JosefaNava <@t> texashealth.org Thu Oct 23 07:30:12 2003 From: JosefaNava <@t> texashealth.org (Nava, Josefa) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] looking for B CATENIN and CDX2 antibodies Message-ID: <3B1A15523787D71197170000E867C0A20120AF87@PHDEX04> Hello Everyone, Where can I order these Antibodies, B CATENIN and CDX2 that will work best for Ventana Immunostainer ? Any help is greatly appreciated. Thanks. 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 have received this message in error, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From georgecole <@t> ev1.net Thu Oct 23 10:07:10 2003 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] SO WHAT DO WE DO?? Message-ID: <000001c39977$561bdb50$0b4dbad0@hppav> Everyday the Histonet presents a parade of new methods--- alternate methods---news about what works----news about what doesn't work. What would a good study of the work practices of us histotechs find looking at----The Great BATTLE???:---In this corner, HABIT, in this corner IMPROVEMENT---- HABIT sings siren songs trying to woo us to bypass IMPROVEMENT----EXCELLENCE CAN'T WAIT TO SEE WHAT HAPPENS. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031023/0ab2d64f/attachment.htm From funderwood <@t> mcohio.org Thu Oct 23 09:39:50 2003 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] MSB dyes Message-ID: Hi Ronnie. acid yellow 24 is available from acros organics(a division of fisher, 1.800.766.7000) catalog #18949.0250 acid blue 93, catalog # M6900; acid red 44, catalog #0644 are available from sigma 1.800.325.3010 Fred >>> "Houston, Ronnie" 10/22/03 11:56AM >>> Hi Folks, I know a lot of you will be in Kentucky just now, but I need your assistance. I'm looking for a source, in the USA, of the following dyes for a MSB stain for fibrin. So far I haven't had much luck, only locating Brilliant crystal scarlet: Martius yellow (acid yellow 24) CI 10315 Brilliant crystal scarlet (acid red 44) CI 16250 Methyl blue (acid blue 93) CI 42780 Thanks Ronnie Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BOULIANNEP <@t> preclin.com Thu Oct 23 11:13:57 2003 From: BOULIANNEP <@t> preclin.com (PATRICE BOULIANNE) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] zinc fixative Message-ID: Hi everybody! I want to know if after a fixation with zinc fixative(for IHC), we can process the tissue in a regular formalin program (paraffin impregnation)? Should I change the first bath (fromalin) for the zinc fixative (with formaline). Is it corrosive for the processor? And should I rince the tissue in tap water before the processing to release zinc residue? Thanks for your help! Patrice Boulianne Histopathotechnologist RT, Team Leader Histopathology Lab Pre-Clinical Research Intl. Inc. 450-973-2240 ext. 1625 email: bouliannep@preclin.com -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031023/4bcfcae8/attachment.htm From gudrun.lang <@t> aon.at Thu Oct 23 11:14:52 2003 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] IHC-controls Message-ID: <013101c39980$ca5493f0$0d12a8c0@SERVER> Dear histonetter, my workmate, who does the IHC on ventana nexes, has a question about using controls. Do you allways use negativ and positiv controls with each probe (or with each antibody), if you do the IHC also on a automated stainer? greetings Gundi Lang general hospital, Linz -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031023/ffea8b0e/attachment.htm From Ronnie_Houston <@t> bshsi.com Thu Oct 23 11:58:03 2003 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] CD 25 Message-ID: <530361BF03351B4CAE5270A05D3037B5FE04C8@bsrexms01.BSHSIR.COM> Is anyone using CD25 on formalin-fixed, paraffin- embedded human tissue? What clone and source are available? Thanks Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 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 mari.ann.mailhiot <@t> leica-microsystems.com Thu Oct 23 12:23:36 2003 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:22:04 2005 Subject: [Histonet] zinc fixative Message-ID: Patrice If your tissue is fixed in zinc formalin, the formulation should be with zinc sulfate and not zinc chloride because zinc chloride is corrosive to metal parts on enclosed tissue processors. In most instances zinc formalin can be used on an enclosed processor again as long as it is made from zinc sulfate. There is no need to follow with NB formalin on the processor. Freida Carson has a reference to zinc formalin in her book Histotechnology - A Self - Instruction Text. Hope this helps. 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 "PATRICE BOULIANNE" To: Sent by: cc: histonet-admin@lists.utsouth Subject: [Histonet] zinc fixative western.edu 10/23/2003 11:13 AM Hi everybody! ??? I want to know? if after a fixation with zinc fixative(for IHC), we can?process the tissue in a regular?formalin program (paraffin impregnation)? Should I change the first bath (fromalin) for?the zinc fixative (with formaline). Is it corrosive for the processor? And should I rince the tissue in tap water before the processing to release zinc residue? Thanks for your help! Patrice Boulianne ?Histopathotechnologist RT, Team Leader ?Histopathology ?Lab Pre-Clinical Research Intl. Inc. ?450-973-2240 ext. 1625 ?email: bouliannep@preclin.com ________________________________________________________________________ This email has been scanned for all viruses by the MessageLabs Email Security System. For more information on a proactive email security service working around the clock, around the globe, visit http://www.messagelabs.com ________________________________________________________________________ From jianqingzheng <@t> hotmail.com Thu Oct 23 12:25:54 2003 From: jianqingzheng <@t> hotmail.com (jianqing zheng) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] help on immunohistochemitry of Guinea Pig anti-Double Cortin Message-ID: Hi,All, I have trouble with Guinea Pig anti-Double Cortin immunohistochemistry on paraffin mouse brain tissue.Any suggetions would be greatly appreciated. Jianqing Zheng MD (706)7362173(H) (706)7214797(L) If a man be gracious and courteous to strangers, it shows he is a citizen of the world, and that his heart is no island cut off from other lands, but a continent that joins them. - Sir Francis Bacon _________________________________________________________________ Send instant messages to anyone on your contact list with MSN Messenger 6.0. Try it now FREE! http://msnmessenger-download.com From GDawson <@t> Milw.Dynacare.com Thu Oct 23 12:23:51 2003 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Beta Catenin Message-ID: Josie, I use the Beta Catenin from BD Biosciences (Cat. #610153). It works well for me but I use the DAKO autostainer rather than Ventana. Good Luck, Glen Dawson From mark.lewis <@t> thermo.com Thu Oct 23 13:54:11 2003 From: mark.lewis <@t> thermo.com (mark.lewis@thermo.com) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Co-Path Message-ID: Hello everyone ! For those of you who were at NSH, I hope you learned something as well as had a good time ! I have a question. What is the difference between the Mysis and Cerner version of CO-Path ? Your response is greatly appreciated. Thanks !! Mark Mark Lewis Product Specialist Anatomical Pathology Clinical Diagnostics Thermo Electron Corporation (412) 747-4013 (412) 788-1097 E-mail: mark.lewis@thermo.com From amosbrooks <@t> earthlink.net Thu Oct 23 14:26:30 2003 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Re: Histonet digest, Vol 1 #99 - 20 msgs In-Reply-To: <20031023170001.9483.74606.Mailman@swlx167.swmed.edu> References: <20031023170001.9483.74606.Mailman@swlx167.swmed.edu> Message-ID: <6.0.0.22.0.20031023152451.029abec0@127.0.0.1> Ann, Mesothelioma works fine. We were using Wilms' as a control and one of the pathologists actually asked us to use mesothelioma instead. Amos At 01:00 PM 10/23/03, you wrote: >Hi Histonetters, > >I am working up Wilms' Tumor (WT1) - and wondering what other labs are >using for control. Unfortunately, a Wilms tumor is not an option for >me. Anyone using a mesothelioma or some other tissue for a control with >this antibody? >Thanks in advance. > >Ann > >Ann Maruska >Fairview-University Medical Center >Mpls. MN 55454 >amarusk1@fairview.org >612-273-9119 From japoteete <@t> saintfrancis.com Thu Oct 23 14:41:56 2003 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Re: Histonet digest, Vol 1 #99 - 20 msgs Message-ID: Ann, please let me know how you do with that antibody. We have tried everything in the world and the thing is still not consistent, so my docs gave up on it. We're using the WT-1 from DAKO, and even had a couple of visits from their specialist. No luck. Jacquie Poteete MT(ASCP), Lead Technologist, IHC Lab Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com > -----Original Message----- > From: Amos Brooks [SMTP:amosbrooks@earthlink.net] > Sent: Thursday, October 23, 2003 2:27 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Re: Histonet digest, Vol 1 #99 - 20 msgs > > Ann, > Mesothelioma works fine. We were using Wilms' as a control and > one > of the pathologists actually asked us to use mesothelioma instead. > Amos > > > At 01:00 PM 10/23/03, you wrote: > >Hi Histonetters, > > > >I am working up Wilms' Tumor (WT1) - and wondering what other labs are > >using for control. Unfortunately, a Wilms tumor is not an option for > >me. Anyone using a mesothelioma or some other tissue for a control with > >this antibody? > >Thanks in advance. > > > >Ann > > > >Ann Maruska > >Fairview-University Medical Center > >Mpls. MN 55454 > >amarusk1@fairview.org > >612-273-9119 > > > _______________________________________________ > 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 akwilliams75 <@t> hotmail.com Thu Oct 23 15:01:29 2003 From: akwilliams75 <@t> hotmail.com (kevin williams) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] (no subject) Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031023/bd60209a/attachment.htm From jlambrey <@t> hotmail.com Thu Oct 23 16:40:44 2003 From: jlambrey <@t> hotmail.com (Julien Lambrey de Souza) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] bone and cartilage staining on mounted sections Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031023/a2697e0c/attachment.htm From mrl0627 <@t> mail.ecu.edu Thu Oct 23 17:08:37 2003 From: mrl0627 <@t> mail.ecu.edu (mrl0627@mail.ecu.edu) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] LMD ... IHC sections falling off Message-ID: <26588589.1066946917921.JavaMail.SYSTEM@onestop9> I sometimes lose sections (4 um thick, mounted on superfrost-plus) during the counterstaining-clearing process although they survive immuno and development in DAB. Any thoughts of what I could be doing wrong? It's a heart breaker to lose them at the very end of the process. I am in a university lab and do everything by hand. Thanks, Maureen at ECU -----Original Message----- From: Amos Brooks <amosbrooks@earthlink.net> To: histonet@lists.utsouthwestern.edu Sent: Mon, 20 Oct 2003 16:20:39 -0400 Subject: [Histonet] LMD ... IHC sections falling off Victoria, Firstly try thinner sections 10 um is a bit thick. Next you should use some type of charged, silanized or APES slides. You can either silanize your own slides or buy them from a vendor. Adhesives aren't necessarily good for IHC as they can interfere with the reaction. Amos Brooks >Message: 5 >From: victoria.hahn-stromberg@orebroll.se >To: histonet@lists.utsouthwestern.edu >Date: Mon, 20 Oct 2003 13:20:39 +0200 >Subject: [Histonet] LMD > >Hi! >I=B4m having problems with my LMD slides. I use 10um paraffin embedded >histological sections which I would like to stain immunohistochemically = >with >CAM, but the sections keep falling off, can anyone plesse help = >me??/Victoria > >Victoria Hahn-Str=F6mberg >MSc, PhD student >Kliniken f=F6r Patologi >Universitetssjukhuset =D6rebro >701 85 =D6rebro >tel: 019-6023644 >faxnr:019-6021035 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From brucea <@t> unimelb.edu.au Thu Oct 23 23:24:25 2003 From: brucea <@t> unimelb.edu.au (Bruce Abaloz) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] (no subject) Message-ID: Hello, I am a PhD student at the University of Melbourne, Australia. I am examining the sperm of moths. Moths have 2 types of sperm a nucleated (eupyrene) sperm and an anucleated (apyrene) sperm. I was hoping to find something that would only stain the nucleated sperm so that the two can be easily differentiated. If you think you could help me it would be very much appreciated. Kathryn McNamara Department of Zoology University of Melbourne Victoria, Australia 3010 -- 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 brucea <@t> unimelb.edu.au Thu Oct 23 23:26:58 2003 From: brucea <@t> unimelb.edu.au (Bruce Abaloz) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] OOPS!....Moth Sperm. Message-ID: Hello Everyone, my name is Kathryn Mcnamara & I am a PhD student at the University of Melbourne, Australia. I am examining the sperm of moths. Moths have 2 types of sperm a nucleated (eupyrene) sperm and an anucleated (apyrene) sperm. I was/AM hoping to find something that would only stain the nucleated sperm so that the two can be easily differentiated with the one stain. If you could help me it would be very much appreciated. THANKS in advance, Kathryn McNamara Department of Zoology University of Melbourne Victoria, Australia 3010 -- 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 edmondsj <@t> musc.edu Fri Oct 24 06:54:45 2003 From: edmondsj <@t> musc.edu (edmondsj) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] LMD ... IHC sections falling off In-Reply-To: <26588589.1066946917921.JavaMail.SYSTEM@onestop9> References: <26588589.1066946917921.JavaMail.SYSTEM@onestop9> Message-ID: <934237.1066982085@jbepc.thurmond-gazes.musc.edu> Hello, Try incubating your slides for a longer period before deparaffinizing that may help with the sections washing off. Hope this helps. Joyce Edmonds MUSC Charleston, SC --On Thursday, October 23, 2003 5:08 PM -0500 mrl0627@mail.ecu.edu wrote: > > I sometimes lose sections (4 um thick, mounted on superfrost-plus) > during the counterstaining-clearing process although they survive immuno > and development in DAB. Any thoughts of what I could be doing wrong? > It's a heart breaker to lose them at the very end of the process. I am > in a university lab and do everything by hand. Thanks, Maureen at ECU > -----Original Message----- > From: Amos Brooks <amosbrooks@earthlink.net> > To: histonet@lists.utsouthwestern.edu > Sent: Mon, 20 Oct 2003 16:20:39 -0400 > Subject: [Histonet] LMD ... IHC sections falling off > > Victoria, > Firstly try thinner sections 10 um is a bit thick. Next you > should use some type of charged, silanized or APES slides. You can > either silanize your own slides or buy them from a vendor. Adhesives > aren't necessarily good for IHC as they can interfere with the reaction. > Amos Brooks > > > > >Message: 5 > >From: victoria.hahn-stromberg@orebroll.se > >To: histonet@lists.utsouthwestern.edu > >Date: Mon, 20 Oct 2003 13:20:39 +0200 > >Subject: [Histonet] LMD > > > >Hi! > >I=B4m having problems with my LMD slides. I use 10um paraffin > embedded >histological sections which I would like to stain > immunohistochemically = >with > >CAM, but the sections keep falling off, can anyone plesse help = > >me??/Victoria > > > >Victoria Hahn-Str=F6mberg > >MSc, PhD student > >Kliniken f=F6r Patologi > >Universitetssjukhuset =D6rebro > >701 85 =D6rebro > >tel: 019-6023644 > >faxnr:019-6021035 > > > _______________________________________________ > 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 lesley <@t> vancouverbc.net Fri Oct 24 10:03:07 2003 From: lesley <@t> vancouverbc.net (Lesley Weston) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Tetracycline and decalcification In-Reply-To: Message-ID: Thanks to everyone who replied to my query. I was asking on behalf of someone else, and I guess he'll just have to find some other way. Lesley Weston. on 16/10/2003 3:01 PM, Lesley Weston at lesley@vancouverbc.net wrote: > Does anyone know if decalcifying tetracycline-treated bone affects its > fluorescence? Everything I can find on the subject says that they embedded > undecalcified bone in methacrylate, but is this essential? If it is OK to > decalcify, should one use acid or EDTA? Sorry to be so helpless, but this > doesn't seem to have been covered anywhere. Thanks to anyone who can help. > > Lesley Weston. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Doug.Geddes <@t> lhsc.on.ca Fri Oct 24 10:47:02 2003 From: Doug.Geddes <@t> lhsc.on.ca (Doug Geddes) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] CD23 and BCL-6 on B5 fixed tissue Message-ID: Looking for any information on performance of CD23 and BCL-6 antibodies on B5 fixed tissue, re - antibody suppliers, incubation times, antigen retreival(?). Thanks so much in advance. Doug Geddes BSc, MLT LHSC, London ON, Canada From amosbrooks <@t> earthlink.net Fri Oct 24 12:46:00 2003 From: amosbrooks <@t> earthlink.net (amosbrooks) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Re: Histonet digest, Vol 1 #100 - 15 msgs Message-ID: <12836864.1067017560984.JavaMail.Administrator@atp> Ronnie, We use Novocastra's CD25 (NCL CD25 305) at 1:100. We pretreat with hot citrate buffer (HIER) and detect with Envision. Amos Brooks From: "Houston, Ronnie" To: "Histonet (E-mail)" Date: Thu, 23 Oct 2003 12:58:03 -0400 Subject: [Histonet] CD 25 Is anyone using CD25 on formalin-fixed, paraffin- embedded human tissue? What clone and source are available? Thanks Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 ronnie_houston@bshsi.com From settembr <@t> umdnj.edu Fri Oct 24 13:09:07 2003 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] CD 25 Message-ID: Hello Ronnie, I use CD25 (Interleukin-2 Receptor) from Novocastra Laboratories which is distributed by Vector Laboratories in Burlingame, CA. I use in on FFPE human tissue. Their clone is 4C9. Their catalog number is NCL-CD25-305. Their spec sheet recommends a tonsil for a positive control. I use it at 1:50 with Dako's Target Retrieval Solution. Good Luck. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> "Houston, Ronnie" 10/23/2003 9:58:03 AM >>> Is anyone using CD25 on formalin-fixed, paraffin- embedded human tissue? What clone and source are available? Thanks Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Fri Oct 24 13:14:46 2003 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Moth Sperm. In-Reply-To: References: Message-ID: <3F996C16.2010000@umdnj.edu> Can you see the nucleus? If so, I don't understand the problem. Perhaps you are trying to stain the whole nucleated sperm one color and the whole anculeate sperm another? Otherwise, just stain for something found in a nucleus and not in the cytoplasm. Geoff Bruce Abaloz wrote: >Hello Everyone, >my name is Kathryn Mcnamara & I am a PhD student at the University of Melbourne, Australia. I am examining the sperm of moths. Moths have 2 types of sperm a nucleated (eupyrene) sperm and an anucleated (apyrene) sperm. >I was/AM hoping to find something that would only stain the nucleated sperm so that the two can be easily differentiated with the one stain. If you could help me it would be very much appreciated. THANKS in advance, > >Kathryn McNamara > >Department of Zoology >University of Melbourne >Victoria, Australia 3010 > > > > -- -- ********************************************** 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 JCarpenter764 <@t> aol.com Fri Oct 24 13:23:51 2003 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] can someone please explain lyses Message-ID: <42.3fe12184.2ccac837@aol.com> -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031024/e328b670/attachment.htm From barbara.wright2 <@t> dnax.org Fri Oct 24 14:20:12 2003 From: barbara.wright2 <@t> dnax.org (Wright, Barbara (SPRI 2)) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] (no subject) Message-ID: <29B25753F6B1D51196110002A589D4440105C783@PALMSG30.us.schp.com> To all my well versed colleagues -- I am in the pursuit of slide labels that are sold in 8.5X11 sheets that can be used in a laser printer. (I haven't decided to use word, excel or file maker pro formats to generate the labels). Any help would be greatly appreciated. Thanks, Barb Barbara Wright Scientist II Exp. Pathology & Pharmacology DNAX 901 California Avenue Palo Alto, CA 94304-1104 barbara.wright2@dnax.org ********************************************************************* 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. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031024/b5a1486a/attachment.htm From GDawson <@t> Milw.Dynacare.com Fri Oct 24 15:01:36 2003 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] CD25 Message-ID: Ronnie, I use CD25 (clone Tu69) from Novocastra. Cat.# NCL-CD25-305. It is for FFPE tissues. Good Luck, Glen Dawson From Bonnie.P.Whitaker <@t> uth.tmc.edu Fri Oct 24 16:59:39 2003 From: Bonnie.P.Whitaker <@t> uth.tmc.edu (Bonnie P Whitaker) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] rhodamine auto-fluor. problem Message-ID: Hi All! I trust everyone that went to Louisville had a wonderful time and learned a lot!! I have a question for you guys: A researcher here in OB/GYN is doing rhodamine-labeled fluorescent work (he didn't say what antibody) on mouse fallopian tube... he thought he was having a staining problem, but has determined that his tissue is auto-fluorescing. The tissue is frozen in OCT and fixed in acetone/methanol. What can he do to quench this? Thanks, Bonnie Whitaker University of Texas Medical School at Houston 6431 Fannin Street MSB 2.231 Houston, Texas 77030 Phone 713.500.6792 Fax 713.500.0733 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031024/b71d7b91/attachment.htm From JCarpenter764 <@t> aol.com Fri Oct 24 17:29:08 2003 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] (no subject) Message-ID: <12.37a4e1f5.2ccb01b4@aol.com> can anyone explain to me what lyses is....i have come across this term several times while studying for my exam. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031024/46eb1501/attachment.htm From Barry.R.Rittman <@t> uth.tmc.edu Fri Oct 24 18:40:14 2003 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] (no subject) Message-ID: <566FB0B522443D43AF02D2ADBE35A6F063587A@UTHEVS3.mail.uthouston.edu> Lysis is the process of destruction of substances. Often this term is used in conjunction with the material being lysed e.g. lysis of red blood cells is often described as hemolysis. Autolysis is the process whereby cells "self destruct". Whe tissue is removed from its support in the body, the pH within the cells decreases and one end result is the rupture of organelles known as lysosomes. These organelles contain a variety of enzymes that usually act in a controoled manner to destroy materials and organisms taken into the cell and also other organelles that need to be broken down so that the components can be reused. When the lysosomes rupture the enzymes are released in the cell and start to break down the surrounding cell components. The same result is seen when you leave a piece of meat on a countertop for several days, it ends up being liquified. Hope this helps Barry Rittman -----Original Message----- From: JCarpenter764@aol.com [mailto:JCarpenter764@aol.com] Sent: Fri 10/24/2003 5:29 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] (no subject) can anyone explain to me what lyses is....i have come across this term several times while studying for my exam. From jnocito <@t> satx.rr.com Fri Oct 24 23:13:33 2003 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Reality Check Message-ID: <00e301c39aae$5b3b3620$70494542@satx.rr.com> I want to wish everyone back from the NSH meeting. Hope you had a good time. Maybe I can attend next year. I have this issue that just won't go away and I would like your opinions. In the last three years, I've purchased 4 TBS waterbaths, two of which the heating element went bad. One was three years old, the other about two years. Now, I realize that TBS has a one year warranty on their equipment, but shouldn't waterbaths last longer than 3 years? I paid over $1000 per waterbath and they want $500 a piece to repair them. I have seen Boekel, Lab-Line and Fisher waterbaths that were older than me (we won't go there ok?) Which is to say that I'm much older than 3 years. Has anyone else come across this problem? I'm just waiting for the other 2 waterbaths to breakdown. I could see maybe one waterbath, but 2, with the same problem. I think the odds are just too high. Am I like in outer space or does this bother anyone else? Maybe it's me. I don't know. Oh, as a last comment, we do not suffer from severe power surges either, just in case someone would ask. Take care and thanks again for your input. Joe Nocito BS, HT (ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, Texas From hadi83 <@t> comcast.net Fri Oct 24 23:12:16 2003 From: hadi83 <@t> comcast.net (Hadi Yaziji) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] CD23 and BCL-6 on B5 fixed tissue In-Reply-To: Message-ID: <6B52F199-06A1-11D8-92CA-00039378E76A@comcast.net> Dear Doug, B5 fixative destroys the signal of many lymphoid markers. Please advise your pathologists that there's no reason to use it on lymphoid cases. Best regards, Hadi Yaziji, M.D. PhenoPath Laboratories On Friday, October 24, 2003, at 08:47 AM, Doug Geddes wrote: > Looking for any information on performance of CD23 and BCL-6 antibodies > on B5 fixed tissue, re - antibody suppliers, incubation times, antigen > retreival(?). Thanks so much in advance. > > Doug Geddes BSc, MLT > LHSC, London ON, Canada > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From asmith <@t> mail.barry.edu Sat Oct 25 10:59:21 2003 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] (no subject) Message-ID: <494304423C63E246A5CF87A3AEEB577011B5A1@bumail01.barrynet.barry.edu> Lyses is the the third person singular of the verb "to lyse". It means "to cause the breakdown of." E.g., "Complement lyses bacteria," or, "Diastase lyses glycogen." Once in a blue moon, lyses is used as the plural of lysis, which is Latin for dissolving. In English, lysis usually means breakdown. In chemistry, it means the splitting of a compound into two parts. In biology, it means the death of a cell and the disappearance of its parts. Allen A. Smith, Ph.D. Professor of Anatomy School of Graduate Medical Sciences Barry University Miami Shores, FL -----Original Message----- From: JCarpenter764@aol.com [mailto:JCarpenter764@aol.com] Sent: Friday, October 24, 2003 6:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) can anyone explain to me what lyses is....i have come across this term several times while studying for my exam. 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) -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031025/9ee80cb8/attachment.htm From RCHIOVETTI <@t> aol.com Sun Oct 26 11:03:36 2003 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] rhodamine auto-fluor. problemI Message-ID: <44.36fc7206.2ccd5868@aol.com> In a message dated 10/24/2003 3:01:17 PM US Mountain Standard Time, Bonnie.P.Whitaker@uth.tmc.edu writes: > I have a question for you guys: A researcher here in OB/GYN is doing > rhodamine-labeled fluorescent work (he didn't say what antibody) on mouse > fallopian tube... he thought he was having a staining problem, but has > determined that his tissue is auto-fluorescing. The tissue is frozen in OCT > and fixed in acetone/methanol. What can he do to quench this? > Hi Bonnie, Is this a new project, or a recent problem with an ongoing project? Is the autofluorescence in the red wavelengths, or is the investigator perhaps seeing a yellow or green color in the scope? I'm wondering if the scope the investigator is using has been successfully used for rhodamine work before? Fluorescence filter sets are usually constructed in one of two ways: They may have a probe-specific "bandpass" filter in them which will only pass the specific wavelength of the probe (rhodamine, in this case), or a "longpass" filter which will pass the signal from the probe, plus other wavelengths which are close to, but not exactly the same as, the probe. One thing's for sure, if your colleague is looking for red fluorescence and is seeing a yellowish or greenish color through the cube, he may need to use a different filter set in the scope. The manufacturer of the scope or the manufacturer of the filter set should be able to give some guidance here. Hope this is of some help! Bob Chiovetti GTI Microsystems Leica Regional Dealer Desert Southwest Region USA From subratab <@t> bdonline.com Sun Oct 26 13:53:16 2003 From: subratab <@t> bdonline.com (Subratab) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Explain the biochemical basis of 'no counterstain with hematoxylin' Message-ID: <200310261957.h9QJv0wv008505@korotoa.bdonline.com> Dear all I am staining (IHC) rat renal tissue for ED1. When I am using Methacarn-fixed tissue I am not getting any counterstain with hematoxylin. Nuclear positions are showing blank space (ghost-like rounded gaps). Tissue is showing just-yellowish appearance. I have stained the methacarn-fixed slides with hematoxylin without going through my IHC protocol (deparaffinization and hematoxylin). And the staining is OK. So the methacarn-fixed tissue has no problem with hematoxylin. It indicates that my IHC protocol is not campatible with hematoxylin stain when I am using methacarn-fixed tissue. On the otherhand, when I am using paraformaldehyde fixed tissue with the same protocol, I am facing no problem to counterstain with hematoxylin. 1. Can you please explain the biochemical basis? 2. Can the problem be solved by methyl green counterstaining? I am expecting clarification and suggestion from the histonet experts.Following is my IHC protocol: Deparaffinization and rehydration Digestion of protein cross-links by trypsin Microwave exposure in citrate buffer Blocking with 1% non-fat milk Primary Ab (ED1) diluted in PBS (1% BSA + 0.3% Triton x100) 3% H2O2 in methanol AP conjugated polymer (anti-mouse Ig) Fast red with substrate buffer and levamisol (from Dako) Counterstaining with hematoxylin and mounting with permafluor. Washing solution is PBS. (ED1 staining is OK. The problem is with counterstaining) Subrata Biswas. University of Campinas, brazil. From clarke.ian <@t> virgin.net Sun Oct 26 15:47:17 2003 From: clarke.ian <@t> virgin.net (ian clarke) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Re computer program to identify stored tissue and store photographs consent forms etc Message-ID: Hi All, I am looking for a software program that would aid the recording of stored tissue along with photographs and copies of consent forms etc.Does anyone know of such a program or something that might do some of the above tasks? Hope you can help Thanks in advance Ian Clarke Histo/Cyto CAH From lpwenk <@t> covad.net Sun Oct 26 17:59:38 2003 From: lpwenk <@t> covad.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] MSB dyes References: <530361BF03351B4CAE5270A05D3037B5FE04BD@bsrexms01.BSHSIR.COM> Message-ID: <014d01c39c1d$37642340$8732fea9@hppav> We've played around with the MSB, and have found it is easier to substitute the dyes you are currently using for a Masson Trichrome. One of my students did a research project on this, in fact. Instead of the dual nuclear stain of celestine blue and Mayer hematoxylin, use the Weigert iron hematoxylin. Looks great, and you don't have to stop and make up celestine blue. Instead of brilliant crystal scarlet, use whatever red dye you are using for Masson trichrome - ponceau de xylidine, ponceau S, biebrich scarlet, acid fuchsin, etc., or any combination. If you don't use methyl blue in your Masson trichrome, use the aniline blue solution. Or, if you like the green of Gomori trichrome, use the fast green FCF. If fixed in NBF, post-mordant. Use either sat. aq. mercuric chloride in 60 degree C for 1 hour (tradition for the MSB stain), or Bouins similar to any trichrome post-mordant, or use the potassium dichromate-HCl for 10 mins. In other words, do an MSB EXACTLY like you do a Masson Trichrome, but insert the alcoholic Martius yellow. - Post mordant - Weigert hematoxylin - Martius yellow - red dye - phosphotungstic acid step -blue or green dye It will look exactly like a Masson trichrome, except that the RBCs will be yellow, not red. This helps when fibrin is mixed in with RBCs. Peggy Wenk William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Houston, Ronnie" To: Sent: Wednesday, October 22, 2003 10:56 AM Subject: [Histonet] MSB dyes > > Hi Folks, > > I know a lot of you will be in Kentucky just now, but I need your > assistance. I'm looking for a source, in the USA, of the following dyes for > a MSB stain for fibrin. So far I haven't had much luck, only locating > Brilliant crystal scarlet: > > Martius yellow (acid yellow 24) CI 10315 > Brilliant crystal scarlet (acid red 44) CI 16250 > Methyl blue (acid blue 93) CI 42780 > > Thanks > Ronnie > > Ronnie Houston > Regional Histology Operations Manager > Bon Secours HealthPartners Laboratories > 5801 Bremo Road > Richmond, VA 23226 > (804) 287 7972 > 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. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From chris.jones <@t> adelaide.edu.au Sun Oct 26 19:15:41 2003 From: chris.jones <@t> adelaide.edu.au (Chris Jones) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] (no subject) Message-ID: <6.0.0.22.1.20031027113612.02531e28@mail.staff.adelaide.edu.au> Dear all, I am hoping that someone can be of assistance. I am trying to fix embryonic chick limb buds, to preserve as much as possible the spatial relations of the cells. A Karnovsky-like fixation (glutaraldehyde + paraformaldehyde, postfixed in osmium tetroxide) fixes really well, but apparently Karnovsky fixatives induce a fair bit of shrinkage. A 'better' fixative ie induces less shrinkage (osmium + glutaraldehyde as primary fixatives) has so far given lesser quality fixation. Does anybody know of a modification we can make (with confidence) to the osmium/glut fix that will improve things? Or does postfixation in osmium induce less shrinkage anyway? I will be most appreciative of any nibbles. Many thanks, Chris. ============================= Christopher Jones, PhD Department of Anatomical Sciences University of Adelaide ADELAIDE SOUTH AUSTRALIA 5005 Ph. +61 8 8303 4526 Fax. +61 8 8303 4398 www.health.adelaide.edu.au/anat ============================= CRICOS Provider Number 00123M ----------------------------------------------------------- This email message is intended only for the addressee(s) and contains information that may be confidential and/or copyright. If you are not the intended recipient please notify the sender by reply email and immediately delete this email. Use, disclosure or reproduction of this email by anyone other than the intended recipient(s) is strictly prohibited. No representation is made that this email or any attachments are free of viruses. Virus scanning is recommended and is the responsibility of the recipient. From ctsmolde <@t> capeheart.uct.ac.za Mon Oct 27 05:05:15 2003 From: ctsmolde <@t> capeheart.uct.ac.za (Jenny Molde) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Getting stented sections to stay on slides. Message-ID: Hi all I have posted this question before and now I am beside myself. I cut 5um sections of stented arteries and mount them on super frost ++ slides. I have tried other slides like Apes, polylysine etc but still have no luck. I have floated out at 57*c and tried 70*c distilled water baths, then floated onto acetone as well but still no luck. I have incubated at 37*c overnight, 10 minutes at 60*c , 70*c for 2 hours but to no avail. I have had it as my sections are really good and all I do now is lose them. Ever felt that way. When I first started doing this work I got them to stay on this way ie pick up on coated slides, incubate at 37*c overnight and deresin in 2x changes of warm xylene at 37*C 15 minutes each. worked for a while. Is there anyone out there experiencing the same kind of problems? Many thanks. By the way I am using Mma Neil Hand`s method. Jenny Molde Cardiovascular Research Unit University of Cape Town South Africa. From akwilliams75 <@t> hotmail.com Mon Oct 27 07:58:35 2003 From: akwilliams75 <@t> hotmail.com (kevin williams) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Alcoholic eosin with floxien Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031027/08edbf87/attachment.htm From JCarpenter764 <@t> aol.com Mon Oct 27 08:32:54 2003 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] (no subject) Message-ID: <71.369a1b95.2cce8696@aol.com> what is the difference between the anhydrous compound and the trihydrate compound? -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031027/b0e64984/attachment.htm From JCarpenter764 <@t> aol.com Mon Oct 27 08:35:45 2003 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] (no subject) Message-ID: <123.26c65b9b.2cce8741@aol.com> can anyone explain eosinophil to me -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031027/3f8152a2/attachment.htm From DKUMISKI <@t> mail.mcg.edu Mon Oct 27 08:54:08 2003 From: DKUMISKI <@t> mail.mcg.edu (Donna Kumiski) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] (no subject) Message-ID: slide labels that are sold in 8.5X11 sheets Diversified Biotech 1-800-796-9199 www.divbio.com cat # MISL-1000 Donna ---------------------------------------------- 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 ----------------------------------------------- >>> "Wright, Barbara (SPRI 2)" 10/24/2003 3:20:12 PM >>> To all my well versed colleagues -- I am in the pursuit of slide labels that are sold in 8.5X11 sheets that can be used in a laser printer. (I haven't decided to use word, excel or file maker pro formats to generate the labels). Any help would be greatly appreciated. Thanks, Barb Barbara Wright Scientist II Exp. Pathology & Pharmacology DNAX 901 California Avenue Palo Alto, CA 94304-1104 barbara.wright2@dnax.org ********************************************************************* 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 grantgd <@t> comcast.net Mon Oct 27 08:58:49 2003 From: grantgd <@t> comcast.net (Gordon Grant) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] RE: Digital Photomicroscopy In-Reply-To: <20031025170001.17825.90458.Mailman@swlx167.swmed.edu> Message-ID: <000601c39c9a$d48098c0$0200a8c0@ggrant> Hello histonetters, I have an Olympus BH-2 microscope fitted with a real nice PM10AD Olympus film camera. I have been real happy with the set up as I take a lot of publication photos. My problem is digital is now the way to go. I borrowed a Coolpix MDC lens adapter with a coolpix and Olympus C-5050 camera. I not very pleased with the results. Does anyone have a recommendation for me. Best Gordon Grant -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Saturday, October 25, 2003 10:00 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet digest, Vol 1 #101 - 12 msgs 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-admin@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: Histonet digest, Vol 1 #100 - 15 msgs (amosbrooks) 2. Re: CD 25 (Dana Settembre) 3. Re: Moth Sperm. (Geoff McAuliffe) 4. can someone please explain lyses (JCarpenter764@aol.com) 5. (no subject) (Wright, Barbara (SPRI 2)) 6. CD25 (Dawson, Glen) 7. rhodamine auto-fluor. problem (Bonnie P Whitaker) 8. (no subject) (JCarpenter764@aol.com) 9. RE: (no subject) (Barry R Rittman) 10. Reality Check (Joe Nocito) 11. Re: CD23 and BCL-6 on B5 fixed tissue (Hadi Yaziji) 12. RE: (no subject) (Smith, Allen) --__--__-- Message: 1 Date: Fri, 24 Oct 2003 10:46:00 -0700 (PDT) From: amosbrooks Reply-To: amosbrooks@earthlink.net To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Histonet digest, Vol 1 #100 - 15 msgs Ronnie, We use Novocastra's CD25 (NCL CD25 305) at 1:100. We pretreat with hot citrate buffer (HIER) and detect with Envision. Amos Brooks From: "Houston, Ronnie" To: "Histonet (E-mail)" Date: Thu, 23 Oct 2003 12:58:03 -0400 Subject: [Histonet] CD 25 Is anyone using CD25 on formalin-fixed, paraffin- embedded human tissue? What clone and source are available? Thanks Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 ronnie_houston@bshsi.com --__--__-- Message: 2 Date: Fri, 24 Oct 2003 14:09:07 -0400 From: Dana Settembre To: Ronnie_Houston@bshsi.com, histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] CD 25 Hello Ronnie, I use CD25 (Interleukin-2 Receptor) from Novocastra Laboratories which is distributed by Vector Laboratories in Burlingame, CA. I use in on FFPE human tissue. Their clone is 4C9. Their catalog number is NCL-CD25-305. Their spec sheet recommends a tonsil for a positive control. I use it at 1:50 with Dako's Target Retrieval Solution. Good Luck. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> "Houston, Ronnie" 10/23/2003 9:58:03 AM >>> Is anyone using CD25 on formalin-fixed, paraffin- embedded human tissue? What clone and source are available? Thanks Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --__--__-- Message: 3 Date: Fri, 24 Oct 2003 14:14:46 -0400 From: Geoff McAuliffe To: Bruce Abaloz Cc: HistoNet , histonet-admin@lists.utsouthwestern.edu Subject: Re: [Histonet] Moth Sperm. Can you see the nucleus? If so, I don't understand the problem. Perhaps you are trying to stain the whole nucleated sperm one color and the whole anculeate sperm another? Otherwise, just stain for something found in a nucleus and not in the cytoplasm. Geoff Bruce Abaloz wrote: >Hello Everyone, >my name is Kathryn Mcnamara & I am a PhD student at the University of Melbourne, Australia. I am examining the sperm of moths. Moths have 2 types of sperm a nucleated (eupyrene) sperm and an anucleated (apyrene) sperm. >I was/AM hoping to find something that would only stain the nucleated sperm so that the two can be easily differentiated with the one stain. If you could help me it would be very much appreciated. THANKS in advance, > >Kathryn McNamara > >Department of Zoology >University of Melbourne >Victoria, Australia 3010 > > > > -- -- ********************************************** 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 ********************************************** --__--__-- Message: 4 From: JCarpenter764@aol.com Date: Fri, 24 Oct 2003 14:23:51 EDT To: histonet@lists.utsouthwestern.edu Subject: [Histonet] can someone please explain lyses -------------------------------1067019831 Content-Type: text/plain; charset="US-ASCII" Content-Transfer-Encoding: 7bit -------------------------------1067019831 Content-Type: text/html; charset="US-ASCII" Content-Transfer-Encoding: quoted-printable -------------------------------1067019831-- --__--__-- Message: 5 From: "Wright, Barbara (SPRI 2)" To: "'histonet@pathology.swmed.edu'" Date: Fri, 24 Oct 2003 15:20:12 -0400 Subject: [Histonet] (no subject) This message is in MIME format. Since your mail reader does not understand this format, some or all of this message may not be legible. ------_=_NextPart_001_01C39A63.D8B02C50 Content-Type: text/plain To all my well versed colleagues -- I am in the pursuit of slide labels that are sold in 8.5X11 sheets that can be used in a laser printer. (I haven't decided to use word, excel or file maker pro formats to generate the labels). Any help would be greatly appreciated. Thanks, Barb Barbara Wright Scientist II Exp. Pathology & Pharmacology DNAX 901 California Avenue Palo Alto, CA 94304-1104 barbara.wright2@dnax.org ********************************************************************* 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. ------_=_NextPart_001_01C39A63.D8B02C50 Content-Type: text/html Content-Transfer-Encoding: quoted-printable

To all my well versed colleagues --

I am in the pursuit of slide labels that a= re sold in 8.5X11 sheets that can be used in a laser printer.  (I have= n't decided to use word, excel or file maker pro formats to generate the la= bels).  Any help would be greatly appreciated.

Thanks,
Barb


Barbara Wright
Scientist II
Exp. Pathology &am= p; Pharmacology
DNAX
901 California Ave= nue
Palo Alto, CA 9430= 4-1104
barbara.wright2@dn= ax.org



*********************************************************************
This message and any attachments are solely for the intended recipient. If = you are not the intended recipient, disclosure, copying, use or distributio= n of the information included in this message is prohibited -- Please immed= iately and permanently delete.
 ------_=_NextPart_001_01C39A63.D8B02C50-- --__--__-- Message: 6 From: "Dawson, Glen" To: histonet@lists.utsouthwestern.edu Date: Fri, 24 Oct 2003 15:01:36 -0500 Subject: [Histonet] CD25 Ronnie, I use CD25 (clone Tu69) from Novocastra. Cat.# NCL-CD25-305. It is for FFPE tissues. Good Luck, Glen Dawson --__--__-- Message: 7 Reply-To: From: "Bonnie P Whitaker" To: "histonet" Date: Fri, 24 Oct 2003 16:59:39 -0500 Subject: [Histonet] rhodamine auto-fluor. problem This is a multi-part message in MIME format. ------=_NextPart_000_0000_01C39A50.36511310 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: 7bit Hi All! I trust everyone that went to Louisville had a wonderful time and learned a lot!! I have a question for you guys: A researcher here in OB/GYN is doing rhodamine-labeled fluorescent work (he didn't say what antibody) on mouse fallopian tube... he thought he was having a staining problem, but has determined that his tissue is auto-fluorescing. The tissue is frozen in OCT and fixed in acetone/methanol. What can he do to quench this? Thanks, Bonnie Whitaker University of Texas Medical School at Houston 6431 Fannin Street MSB 2.231 Houston, Texas 77030 Phone 713.500.6792 Fax 713.500.0733 ------=_NextPart_000_0000_01C39A50.36511310 Content-Type: text/html; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable
Hi All! =
 
I trust everyone = that went to=20 Louisville had a wonderful time and learned a lot!!
 
I have a question = for you=20 guys:  A researcher here in OB/GYN is doing rhodamine-labeled = fluorescent=20 work (he didn't say what antibody) on mouse fallopian tube... he thought = he was=20 having a staining problem, but has determined that his tissue is=20 auto-fluorescing.  The tissue is frozen in OCT and fixed in=20 acetone/methanol.  What can he do to quench = this?
 
Thanks,

Bonnie Whitaker
University of Texas = Medical=20 School at Houston
6431 Fannin Street
MSB 2.231
Houston, = Texas =20 77030
Phone 713.500.6792
Fax 713.500.0733

 

------=_NextPart_000_0000_01C39A50.36511310-- --__--__-- Message: 8 From: JCarpenter764@aol.com Date: Fri, 24 Oct 2003 18:29:08 EDT To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) -------------------------------1067034548 Content-Type: text/plain; charset="US-ASCII" Content-Transfer-Encoding: 7bit can anyone explain to me what lyses is....i have come across this term several times while studying for my exam. -------------------------------1067034548 Content-Type: text/html; charset="US-ASCII" Content-Transfer-Encoding: quoted-printable can anyone explain to me what lyses is....i have come across this term se= veral times while studying for my exam. -------------------------------1067034548-- --__--__-- Message: 9 Date: Fri, 24 Oct 2003 18:40:14 -0500 From: "Barry R Rittman" To: , "histonet" Subject: RE: [Histonet] (no subject) THlzaXMgaXMgdGhlIHByb2Nlc3Mgb2YgZGVzdHJ1Y3Rpb24gb2Ygc3Vic3RhbmNlcy4gT2Z0 ZW4g dGhpcyB0ZXJtIGlzIHVzZWQgaW4gY29uanVuY3Rpb24gd2l0aCB0aGUgbWF0ZXJpYWwgYmVp bmcg bHlzZWQgZS5nLiBseXNpcyBvZiByZWQgYmxvb2QgY2VsbHMgaXMgb2Z0ZW4gZGVzY3JpYmVk IGFz IGhlbW9seXNpcy4NCkF1dG9seXNpcyBpcyB0aGUgcHJvY2VzcyB3aGVyZWJ5IGNlbGxzICJz ZWxm IGRlc3RydWN0Ii4gV2hlIHRpc3N1ZSBpcyByZW1vdmVkIGZyb20gaXRzIHN1cHBvcnQgaW4g dGhl IGJvZHksIHRoZSBwSCB3aXRoaW4gdGhlIGNlbGxzIGRlY3JlYXNlcyBhbmQgb25lIGVuZCBy ZXN1 bHQgaXMgdGhlIHJ1cHR1cmUgb2Ygb3JnYW5lbGxlcyBrbm93biBhcyBseXNvc29tZXMuIFRo ZXNl IG9yZ2FuZWxsZXMgY29udGFpbiBhIHZhcmlldHkgb2YgZW56eW1lcyB0aGF0IHVzdWFsbHkg YWN0 IGluIGEgY29udHJvb2xlZCBtYW5uZXIgdG8gZGVzdHJveSBtYXRlcmlhbHMgYW5kIG9yZ2Fu aXNt cyB0YWtlbiBpbnRvIHRoZSBjZWxsIGFuZCBhbHNvIG90aGVyIG9yZ2FuZWxsZXMgdGhhdCBu ZWVk IHRvIGJlIGJyb2tlbiBkb3duIHNvIHRoYXQgdGhlIGNvbXBvbmVudHMgY2FuIGJlIHJldXNl ZC4g DQpXaGVuIHRoZSBseXNvc29tZXMgcnVwdHVyZSB0aGUgZW56eW1lcyBhcmUgcmVsZWFzZWQg aW4g dGhlIGNlbGwgYW5kIHN0YXJ0IHRvIGJyZWFrIGRvd24gdGhlIHN1cnJvdW5kaW5nIGNlbGwg Y29t cG9uZW50cy4gVGhlIHNhbWUgcmVzdWx0IGlzIHNlZW4gd2hlbiB5b3UgbGVhdmUgYSBwaWVj ZSBv ZiBtZWF0IG9uIGEgY291bnRlcnRvcCBmb3Igc2V2ZXJhbCBkYXlzLCBpdCBlbmRzIHVwIGJl aW5n IGxpcXVpZmllZC4NCkhvcGUgdGhpcyBoZWxwcw0KQmFycnkgUml0dG1hbg0KDQoJLS0tLS1P cmln aW5hbCBNZXNzYWdlLS0tLS0gDQoJRnJvbTogSkNhcnBlbnRlcjc2NEBhb2wuY29tIFttYWls dG86 SkNhcnBlbnRlcjc2NEBhb2wuY29tXSANCglTZW50OiBGcmkgMTAvMjQvMjAwMyA1OjI5IFBN IA0K CVRvOiBoaXN0b25ldEBsaXN0cy51dHNvdXRod2VzdGVybi5lZHUgDQoJQ2M6IA0KCVN1Ympl Y3Q6 IFtIaXN0b25ldF0gKG5vIHN1YmplY3QpDQoJDQoJDQoJY2FuIGFueW9uZSBleHBsYWluIHRv IG1l IHdoYXQgbHlzZXMgaXMuLi4uaSBoYXZlIGNvbWUgYWNyb3NzIHRoaXMgdGVybSBzZXZlcmFs IHRp bWVzIHdoaWxlIHN0dWR5aW5nIGZvciBteSBleGFtLg0KDQo= --__--__-- Message: 10 From: "Joe Nocito" To: "Histonet" Date: Fri, 24 Oct 2003 21:13:33 -0700 Subject: [Histonet] Reality Check I want to wish everyone back from the NSH meeting. Hope you had a good time. Maybe I can attend next year. I have this issue that just won't go away and I would like your opinions. In the last three years, I've purchased 4 TBS waterbaths, two of which the heating element went bad. One was three years old, the other about two years. Now, I realize that TBS has a one year warranty on their equipment, but shouldn't waterbaths last longer than 3 years? I paid over $1000 per waterbath and they want $500 a piece to repair them. I have seen Boekel, Lab-Line and Fisher waterbaths that were older than me (we won't go there ok?) Which is to say that I'm much older than 3 years. Has anyone else come across this problem? I'm just waiting for the other 2 waterbaths to breakdown. I could see maybe one waterbath, but 2, with the same problem. I think the odds are just too high. Am I like in outer space or does this bother anyone else? Maybe it's me. I don't know. Oh, as a last comment, we do not suffer from severe power surges either, just in case someone would ask. Take care and thanks again for your input. Joe Nocito BS, HT (ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, Texas --__--__-- Message: 11 Date: Fri, 24 Oct 2003 21:12:16 -0700 Cc: To: "Doug Geddes" From: Hadi Yaziji Subject: Re: [Histonet] CD23 and BCL-6 on B5 fixed tissue Dear Doug, B5 fixative destroys the signal of many lymphoid markers. Please advise your pathologists that there's no reason to use it on lymphoid cases. Best regards, Hadi Yaziji, M.D. PhenoPath Laboratories On Friday, October 24, 2003, at 08:47 AM, Doug Geddes wrote: > Looking for any information on performance of CD23 and BCL-6 antibodies > on B5 fixed tissue, re - antibody suppliers, incubation times, antigen > retreival(?). Thanks so much in advance. > > Doug Geddes BSc, MLT > LHSC, London ON, Canada > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > --__--__-- Message: 12 Date: Sat, 25 Oct 2003 11:59:21 -0400 From: "Smith, Allen" To: Cc: Subject: RE: [Histonet] (no subject) This is a multi-part message in MIME format. ------_=_NextPart_001_01C39B10.F4706942 Content-Type: text/plain; charset="us-ascii" Content-Transfer-Encoding: quoted-printable Lyses is the the third person singular of the verb "to lyse". It means "= to cause the breakdown of." E.g., "Complement lyses bacteria," or, "Dia= stase lyses glycogen." Once in a blue moon, lyses is used as the plur= al of lysis, which is Latin for dissolving. In English, lysis usually m= eans breakdown. In chemistry, it means the splitting of a compound into= two parts. In biology, it means the death of a cell and the disappeara= nce of its parts. Allen A. Smith, Ph.D. Professor of Anatomy School= of Graduate Medical Sciences Barry University Miami Shores, FL --= ---Original Message----- From: JCarpenter764@aol.com [mailto:JCarpenter= 764@aol.com] Sent: Friday, October 24, 2003 6:29 PM To: histonet@lis= ts.utsouthwestern.edu Subject: [Histonet] (no subject) can anyo= ne explain to me what lyses is....i have come across this term several t= imes while studying for my exam. = The information transmitted is intended only for the = person or entity to which it is addressed and may contain confidential, a= nd/or privileged material. No confidentiality or privilege is waived or l= ost by any errant transmission. If you receive this message in error, ple= ase immediately delete it and all copies of it from your system and notif= y the sender. E-mail transmission cannot be guaranteed to be secure or e= rror-free as information could be intercepted, corrupted, lost, destroyed= , arrive late or incomplete, or contain viruses. Barry University - Miam= i Shores, FL (http://www.barry.edu) ------_=_NextPart_001_01C39B10.F4706942 Content-Type: text/html; charset="us-ascii" Content-Transfer-Encoding: quoted-printable Message
Lyses is the the third person singular of th= e verb "to lyse".  It means "to cause the breakdown of."  E.g= ., "Complement lyses bacteria," or, "Diastase lyses glycogen." 
Once in a blue moon, lyses is used as the plural of lysis, which is Latin = for dissolving.  In English, lysis usually means breakdown.  = In chemistry, it means the splitting of a compound into two parts. = ; In biology, it means the death of a cell and the disappearance of its= parts.
 = ;

A= llen A. Smith, Ph.D.
Professor of Anatomy
School of Graduate Medi= cal Sciences
Barry University
Miami Shores, FL

=
-----Original Message-----
Fr= om: JCarpenter764@aol.com [mailto:JCarpenter764@aol.com]
S= ent: Friday, October 24, 2003 6:29 PM
To: histonet@= lists.utsouthwestern.edu
Subject: [Histonet] (no subject)
can anyone explain to me what lyses is....i have c= ome across this term several times while studying for my exam.


The information transmitted is intended only for the = person or entity to which it is addressed and may contain confidential, a= nd/or privileged material. No confidentiality or privilege is waived or l= ost by any errant transmission. If you receive this message in error, ple= ase immediately delete it and all copies of it from your system and notif= y the sender. E-mail transmission cannot be guaranteed to be secure or er= ror-free as information could be intercepted, corrupted, lost, destroyed,= arrive late or incomplete, or contain viruses.
Barry University - Mia= mi Shores, FL (http://www.barry.edu) =00 ------_=_NextPart_001_01C39B10.F4706942-- --__--__-- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest From bhewlett <@t> cogeco.ca Mon Oct 27 09:07:19 2003 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] RE: Digital Photomicroscopy References: <000601c39c9a$d48098c0$0200a8c0@ggrant> Message-ID: <001301c39c9c$04b6b5a0$6400a8c0@bryaniwx13voft> Gordon, I faced the same dilemma! My solution was to continue using my film camera and purchase a 4000 ppi film scanner. This also gave me the advantage of being able to digitize existing photomicrographs. The results are spectacular! Bryan ----- Original Message ----- From: "Gordon Grant" To: Sent: Monday, October 27, 2003 9:58 AM Subject: [Histonet] RE: Digital Photomicroscopy > Hello histonetters, > I have an Olympus BH-2 microscope fitted with a real nice PM10AD Olympus > film camera. I have been real happy with the set up as I take a lot of > publication photos. My problem is digital is now the way to go. I > borrowed a Coolpix MDC lens adapter with a coolpix and Olympus C-5050 > camera. I not very pleased with the results. Does anyone have a > recommendation for me. > > Best > Gordon Grant > > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of > histonet-request@lists.utsouthwestern.edu > Sent: Saturday, October 25, 2003 10:00 AM > To: histonet@lists.utsouthwestern.edu > Subject: Histonet digest, Vol 1 #101 - 12 msgs > > 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-admin@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: Histonet digest, Vol 1 #100 - 15 msgs (amosbrooks) > 2. Re: CD 25 (Dana Settembre) > 3. Re: Moth Sperm. (Geoff McAuliffe) > 4. can someone please explain lyses (JCarpenter764@aol.com) > 5. (no subject) (Wright, Barbara (SPRI 2)) > 6. CD25 (Dawson, Glen) > 7. rhodamine auto-fluor. problem (Bonnie P Whitaker) > 8. (no subject) (JCarpenter764@aol.com) > 9. RE: (no subject) (Barry R Rittman) > 10. Reality Check (Joe Nocito) > 11. Re: CD23 and BCL-6 on B5 fixed tissue (Hadi Yaziji) > 12. RE: (no subject) (Smith, Allen) > > --__--__-- > > Message: 1 > Date: Fri, 24 Oct 2003 10:46:00 -0700 (PDT) > From: amosbrooks > Reply-To: amosbrooks@earthlink.net > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Re: Histonet digest, Vol 1 #100 - 15 msgs > > Ronnie, > We use Novocastra's CD25 (NCL CD25 305) at 1:100. We pretreat with > hot > citrate buffer (HIER) and detect with Envision. > Amos Brooks > > > > From: "Houston, Ronnie" > To: "Histonet (E-mail)" > Date: Thu, 23 Oct 2003 12:58:03 -0400 > Subject: [Histonet] CD 25 > > > Is anyone using CD25 on formalin-fixed, paraffin- > embedded human tissue? What clone and source are available? > > Thanks > Ronnie Houston > Regional Histology Operations Manager > Bon Secours HealthPartners Laboratories > 5801 Bremo Road > Richmond, VA 23226 > (804) 287 7972 > ronnie_houston@bshsi.com > > > --__--__-- > > Message: 2 > Date: Fri, 24 Oct 2003 14:09:07 -0400 > From: Dana Settembre > To: Ronnie_Houston@bshsi.com, histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] CD 25 > > Hello Ronnie, > I use CD25 (Interleukin-2 Receptor) from Novocastra Laboratories which > is distributed by Vector Laboratories in Burlingame, CA. > I use in on FFPE human tissue. Their clone is 4C9. Their catalog > number is NCL-CD25-305. > Their spec sheet recommends a tonsil for a positive control. > I use it at 1:50 with Dako's Target Retrieval Solution. > Good Luck. > Dana Settembre > University Hospital-UMDNJ > Newark, NJ > > > >>> "Houston, Ronnie" 10/23/2003 9:58:03 AM > >>> > > Is anyone using CD25 on formalin-fixed, paraffin- > embedded human tissue? What clone and source are available? > > Thanks > Ronnie Houston > Regional Histology Operations Manager > Bon Secours HealthPartners Laboratories > 5801 Bremo Road > Richmond, VA 23226 > (804) 287 7972 > 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. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > --__--__-- > > Message: 3 > Date: Fri, 24 Oct 2003 14:14:46 -0400 > From: Geoff McAuliffe > To: Bruce Abaloz > Cc: HistoNet , > histonet-admin@lists.utsouthwestern.edu > Subject: Re: [Histonet] Moth Sperm. > > Can you see the nucleus? If so, I don't understand the problem. Perhaps > you are trying to stain the whole nucleated sperm one color and the > whole anculeate sperm another? Otherwise, just stain for something found > > in a nucleus and not in the cytoplasm. > > Geoff > > Bruce Abaloz wrote: > > >Hello Everyone, > >my name is Kathryn Mcnamara & I am a PhD student at the University of > Melbourne, Australia. I am examining the sperm of moths. Moths have 2 > types of sperm a nucleated (eupyrene) sperm and an anucleated (apyrene) > sperm. > >I was/AM hoping to find something that would only stain the nucleated > sperm so that the two can be easily differentiated with the one stain. > If you could help me it would be very much appreciated. THANKS in > advance, > > > >Kathryn McNamara > > > >Department of Zoology > >University of Melbourne > >Victoria, Australia 3010 > > > > > > > > > > -- > -- > ********************************************** > 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 > ********************************************** > > > > > > --__--__-- > > Message: 4 > From: JCarpenter764@aol.com > Date: Fri, 24 Oct 2003 14:23:51 EDT > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] can someone please explain lyses > > > -------------------------------1067019831 > Content-Type: text/plain; charset="US-ASCII" > Content-Transfer-Encoding: 7bit > > > > -------------------------------1067019831 > Content-Type: text/html; charset="US-ASCII" > Content-Transfer-Encoding: quoted-printable > > > charse= > t=3Dutf-8"> > > #fffff= > f"> > > -------------------------------1067019831-- > > > --__--__-- > > Message: 5 > From: "Wright, Barbara (SPRI 2)" > To: "'histonet@pathology.swmed.edu'" > Date: Fri, 24 Oct 2003 15:20:12 -0400 > Subject: [Histonet] (no subject) > > This message is in MIME format. Since your mail reader does not > understand > this format, some or all of this message may not be legible. > > ------_=_NextPart_001_01C39A63.D8B02C50 > Content-Type: text/plain > > To all my well versed colleagues -- > > I am in the pursuit of slide labels that are sold in 8.5X11 sheets that > can > be used in a laser printer. (I haven't decided to use word, excel or > file > maker pro formats to generate the labels). Any help would be greatly > appreciated. > > Thanks, > Barb > > > Barbara Wright > Scientist II > Exp. Pathology & Pharmacology > DNAX > 901 California Avenue > Palo Alto, CA 94304-1104 > barbara.wright2@dnax.org > > > > ********************************************************************* > 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. > > > > ------_=_NextPart_001_01C39A63.D8B02C50 > Content-Type: text/html > Content-Transfer-Encoding: quoted-printable > > > > > charset=3Dus-ascii"> > 5.5.2653.12"> > > > > >

To all my well versed colleagues > -- >

> >

I am in the pursuit of slide labels > that a= > re sold in 8.5X11 sheets that can be used in a laser printer.  (I > have= > n't decided to use word, excel or file maker pro formats to generate the > la= > bels).  Any help would be greatly appreciated.

> >

Thanks, >
Barb >

>
> >

Barbara > Wright T> >
Scientist > II > >
Exp. Pathology > &am= > p; Pharmacology >
DNAX >
901 California > Ave= > nue >
Palo Alto, CA > 9430= > 4-1104 >
ITC">barbara.wright2@dn= > ax.org >

> >
>
> *********************************************************************
> > This message and any attachments are solely for the intended recipient. > If = > you are not the intended recipient, disclosure, copying, use or > distributio= > n of the information included in this message is prohibited -- Please > immed= > iately and permanently delete.
> > > > ------_=_NextPart_001_01C39A63.D8B02C50-- > > > --__--__-- > > Message: 6 > From: "Dawson, Glen" > To: histonet@lists.utsouthwestern.edu > Date: Fri, 24 Oct 2003 15:01:36 -0500 > Subject: [Histonet] CD25 > > Ronnie, > > I use CD25 (clone Tu69) from Novocastra. Cat.# NCL-CD25-305. It is for > FFPE tissues. > > Good Luck, > > Glen Dawson > > > --__--__-- > > Message: 7 > Reply-To: > From: "Bonnie P Whitaker" > To: "histonet" > Date: Fri, 24 Oct 2003 16:59:39 -0500 > Subject: [Histonet] rhodamine auto-fluor. problem > > This is a multi-part message in MIME format. > > ------=_NextPart_000_0000_01C39A50.36511310 > Content-Type: text/plain; > charset="iso-8859-1" > Content-Transfer-Encoding: 7bit > > Hi All! > > I trust everyone that went to Louisville had a wonderful time and > learned a > lot!! > > I have a question for you guys: A researcher here in OB/GYN is doing > rhodamine-labeled fluorescent work (he didn't say what antibody) on > mouse > fallopian tube... he thought he was having a staining problem, but has > determined that his tissue is auto-fluorescing. The tissue is frozen in > OCT > and fixed in acetone/methanol. What can he do to quench this? > > Thanks, > Bonnie Whitaker > University of Texas Medical School at Houston > 6431 Fannin Street > MSB 2.231 > Houston, Texas 77030 > Phone 713.500.6792 > Fax 713.500.0733 > > > > > ------=_NextPart_000_0000_01C39A50.36511310 > Content-Type: text/html; > charset="iso-8859-1" > Content-Transfer-Encoding: quoted-printable > > > > charset=3Diso-8859-1"> > > >
Hi All! = >
>
size=3D2> 
>
I trust everyone = > that went to=20 > Louisville had a wonderful time and learned a lot!!
>
size=3D2> 
>
I have a question = > for you=20 > guys:  A researcher here in OB/GYN is doing rhodamine-labeled = > fluorescent=20 > work (he didn't say what antibody) on mouse fallopian tube... he thought > = > he was=20 > having a staining problem, but has determined that his tissue is=20 > auto-fluorescing.  The tissue is frozen in OCT and fixed in=20 > acetone/methanol.  What can he do to quench = > this?
>
size=3D2> 
>
size=3D2>Thanks,
>
>

Bonnie Whitaker
University of Texas > = > Medical=20 > School at Houston
6431 Fannin Street
MSB 2.231
Houston, = > Texas =20 > 77030
Phone 713.500.6792
Fax 713.500.0733

>
 
>

> > ------=_NextPart_000_0000_01C39A50.36511310-- > > > > --__--__-- > > Message: 8 > From: JCarpenter764@aol.com > Date: Fri, 24 Oct 2003 18:29:08 EDT > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] (no subject) > > > -------------------------------1067034548 > Content-Type: text/plain; charset="US-ASCII" > Content-Transfer-Encoding: 7bit > > can anyone explain to me what lyses is....i have come across this term > several times while studying for my exam. > > -------------------------------1067034548 > Content-Type: text/html; charset="US-ASCII" > Content-Transfer-Encoding: quoted-printable > > > charse= > t=3Dutf-8"> > > #fffff= > f">can anyone explain to me what lyses is....i have come across this > term se= > veral times while studying for my exam. > > -------------------------------1067034548-- > > > --__--__-- > > Message: 9 > Date: Fri, 24 Oct 2003 18:40:14 -0500 > From: "Barry R Rittman" > To: , > "histonet" > Subject: RE: [Histonet] (no subject) > > THlzaXMgaXMgdGhlIHByb2Nlc3Mgb2YgZGVzdHJ1Y3Rpb24gb2Ygc3Vic3RhbmNlcy4gT2Z0 > ZW4g > dGhpcyB0ZXJtIGlzIHVzZWQgaW4gY29uanVuY3Rpb24gd2l0aCB0aGUgbWF0ZXJpYWwgYmVp > bmcg > bHlzZWQgZS5nLiBseXNpcyBvZiByZWQgYmxvb2QgY2VsbHMgaXMgb2Z0ZW4gZGVzY3JpYmVk > IGFz > IGhlbW9seXNpcy4NCkF1dG9seXNpcyBpcyB0aGUgcHJvY2VzcyB3aGVyZWJ5IGNlbGxzICJz > ZWxm > IGRlc3RydWN0Ii4gV2hlIHRpc3N1ZSBpcyByZW1vdmVkIGZyb20gaXRzIHN1cHBvcnQgaW4g > dGhl > IGJvZHksIHRoZSBwSCB3aXRoaW4gdGhlIGNlbGxzIGRlY3JlYXNlcyBhbmQgb25lIGVuZCBy > ZXN1 > bHQgaXMgdGhlIHJ1cHR1cmUgb2Ygb3JnYW5lbGxlcyBrbm93biBhcyBseXNvc29tZXMuIFRo > ZXNl > IG9yZ2FuZWxsZXMgY29udGFpbiBhIHZhcmlldHkgb2YgZW56eW1lcyB0aGF0IHVzdWFsbHkg > YWN0 > IGluIGEgY29udHJvb2xlZCBtYW5uZXIgdG8gZGVzdHJveSBtYXRlcmlhbHMgYW5kIG9yZ2Fu > aXNt > cyB0YWtlbiBpbnRvIHRoZSBjZWxsIGFuZCBhbHNvIG90aGVyIG9yZ2FuZWxsZXMgdGhhdCBu > ZWVk > IHRvIGJlIGJyb2tlbiBkb3duIHNvIHRoYXQgdGhlIGNvbXBvbmVudHMgY2FuIGJlIHJldXNl > ZC4g > DQpXaGVuIHRoZSBseXNvc29tZXMgcnVwdHVyZSB0aGUgZW56eW1lcyBhcmUgcmVsZWFzZWQg > aW4g > dGhlIGNlbGwgYW5kIHN0YXJ0IHRvIGJyZWFrIGRvd24gdGhlIHN1cnJvdW5kaW5nIGNlbGwg > Y29t > cG9uZW50cy4gVGhlIHNhbWUgcmVzdWx0IGlzIHNlZW4gd2hlbiB5b3UgbGVhdmUgYSBwaWVj > ZSBv > ZiBtZWF0IG9uIGEgY291bnRlcnRvcCBmb3Igc2V2ZXJhbCBkYXlzLCBpdCBlbmRzIHVwIGJl > aW5n > IGxpcXVpZmllZC4NCkhvcGUgdGhpcyBoZWxwcw0KQmFycnkgUml0dG1hbg0KDQoJLS0tLS1P > cmln > aW5hbCBNZXNzYWdlLS0tLS0gDQoJRnJvbTogSkNhcnBlbnRlcjc2NEBhb2wuY29tIFttYWls > dG86 > SkNhcnBlbnRlcjc2NEBhb2wuY29tXSANCglTZW50OiBGcmkgMTAvMjQvMjAwMyA1OjI5IFBN > IA0K > CVRvOiBoaXN0b25ldEBsaXN0cy51dHNvdXRod2VzdGVybi5lZHUgDQoJQ2M6IA0KCVN1Ympl > Y3Q6 > IFtIaXN0b25ldF0gKG5vIHN1YmplY3QpDQoJDQoJDQoJY2FuIGFueW9uZSBleHBsYWluIHRv > IG1l > IHdoYXQgbHlzZXMgaXMuLi4uaSBoYXZlIGNvbWUgYWNyb3NzIHRoaXMgdGVybSBzZXZlcmFs > IHRp > bWVzIHdoaWxlIHN0dWR5aW5nIGZvciBteSBleGFtLg0KDQo= > > > --__--__-- > > Message: 10 > From: "Joe Nocito" > To: "Histonet" > Date: Fri, 24 Oct 2003 21:13:33 -0700 > Subject: [Histonet] Reality Check > > I want to wish everyone back from the NSH meeting. Hope you had a > good > time. Maybe I can attend next year. > I have this issue that just won't go away and I would like your > opinions. > In the last three years, I've purchased 4 TBS waterbaths, two of > which > the heating element went bad. One was three years old, the other about > two > years. Now, I realize that TBS has a one year warranty on their > equipment, > but shouldn't waterbaths last longer than 3 years? I paid over $1000 per > waterbath and they want $500 a piece to repair them. I have seen > Boekel, > Lab-Line and Fisher waterbaths that were older than me (we won't go > there > ok?) Which is to say that I'm much older than 3 years. > Has anyone else come across this problem? I'm just waiting for the > other 2 waterbaths to breakdown. I could see maybe one waterbath, but > 2, > with the same problem. I think the odds are just too high. Am I like > in > outer space or does this bother anyone else? Maybe it's me. I don't > know. > Oh, as a last comment, we do not suffer from severe power surges either, > just in case someone would ask. Take care and thanks again for your > input. > > Joe Nocito BS, HT (ASCP) QIHC > Histology Manager > Pathology Reference Lab > San Antonio, Texas > > > > --__--__-- > > Message: 11 > Date: Fri, 24 Oct 2003 21:12:16 -0700 > Cc: > To: "Doug Geddes" > From: Hadi Yaziji > Subject: Re: [Histonet] CD23 and BCL-6 on B5 fixed tissue > > Dear Doug, > > B5 fixative destroys the signal of many lymphoid markers. Please advise > your pathologists that there's no reason to use it on lymphoid cases. > > Best regards, > Hadi Yaziji, M.D. > PhenoPath Laboratories > > On Friday, October 24, 2003, at 08:47 AM, Doug Geddes wrote: > > > Looking for any information on performance of CD23 and BCL-6 > antibodies > > on B5 fixed tissue, re - antibody suppliers, incubation times, antigen > > retreival(?). Thanks so much in advance. > > > > Doug Geddes BSc, MLT > > LHSC, London ON, Canada > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > --__--__-- > > Message: 12 > Date: Sat, 25 Oct 2003 11:59:21 -0400 > From: "Smith, Allen" > To: > Cc: > Subject: RE: [Histonet] (no subject) > > This is a multi-part message in MIME format. > > ------_=_NextPart_001_01C39B10.F4706942 > Content-Type: text/plain; > charset="us-ascii" > Content-Transfer-Encoding: quoted-printable > > Lyses is the the third person singular of the verb "to lyse". It means > "= > to > cause the breakdown of." E.g., "Complement lyses bacteria," or, "Dia= > stase > lyses glycogen." > Once in a blue moon, lyses is used as the plur= > al of lysis, which is Latin > for dissolving. In English, lysis usually m= > eans breakdown. In chemistry, > it means the splitting of a compound into= > two parts. In biology, it means > the death of a cell and the disappeara= > nce of its parts. > > Allen A. Smith, Ph.D. > Professor of Anatomy > School= > of Graduate Medical Sciences > Barry University > Miami Shores, FL > > --= > ---Original Message----- > From: JCarpenter764@aol.com [mailto:JCarpenter= > 764@aol.com] > Sent: Friday, October 24, 2003 6:29 PM > To: histonet@lis= > ts.utsouthwestern.edu > Subject: [Histonet] (no subject) > > > can anyo= > ne explain to me what lyses is....i have come across this > term several t= > imes while studying for my exam. > > > = > > > The information transmitted is intended only for the = > person or entity to which it is addressed and may contain confidential, > a= > nd/or privileged material. No confidentiality or privilege is waived or > l= > ost by any errant transmission. If you receive this message in error, > ple= > ase immediately delete it and all copies of it from your system and > notif= > y the sender. E-mail transmission cannot be guaranteed to be secure or > e= > rror-free as information could be intercepted, corrupted, lost, > destroyed= > , arrive late or incomplete, or contain viruses. > Barry University - Miam= > i Shores, FL (http://www.barry.edu) > > ------_=_NextPart_001_01C39B10.F4706942 > Content-Type: text/html; > charset="us-ascii" > Content-Transfer-Encoding: quoted-printable > > > AD>Message > t/html; charset=3Dus-ascii"> > e=3DGENERATOR> > ; BACKGROUND-COLOR: #ffffff"> >
T color=3D#0000ff size=3D4>Lyses is the the > third person singular of th= > e verb "to lyse".  It means "to cause the > breakdown of."  E.g= > ., "Complement lyses bacteria," or, "Diastase lyses > glycogen."  FONT>
>
=3D421064715-25102003>Once in a > blue moon, l 715-25102003>yses is used as the > plural of lysis, which is Latin = > for dissolving.  In English, lysis usually > means breakdown.  = > In chemistry, it means the splitting of a compound into > two parts. = > ; In biology, it means the death of a cell and the disappearance > of its= > parts.
>
r=3D#0000ff> class=3D421064715-25102003> = > ;
>
5-25102003> >

A= > llen A. Smith, Ph.D.
Professor of Anatomy
School of > Graduate Medi= > cal Sciences
Barry University
Miami Shores, FL >

= >
>
>
DIV> >
left> face=3DTahoma size=3D2>-----Original Message-----
Fr= > om: > JCarpenter764@aol.com [mailto:JCarpenter764@aol.com]
S= > ent: Friday, > October 24, 2003 6:29 PM
To: > histonet@= > lists.utsouthwestern.edu
Subject: [Histonet] (no > subject) R>
can anyone explain to me what lyses is....i have > c= > ome across this term several times while studying for my exam. > UOTE>


The information transmitted is intended only for the > = > person or entity to which it is addressed and may contain confidential, > a= > nd/or privileged material. No confidentiality or privilege is waived or > l= > ost by any errant transmission. If you receive this message in error, > ple= > ase immediately delete it and all copies of it from your system and > notif= > y the sender. E-mail transmission cannot be guaranteed to be secure or > er= > ror-free as information could be intercepted, corrupted, lost, > destroyed,= > arrive late or incomplete, or contain viruses.
Barry University - > Mia= > mi Shores, FL (http://www.barry.edu) > =00 > ------_=_NextPart_001_01C39B10.F4706942-- > > > > --__--__-- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Reuel.Cornelia <@t> tsrh.org Mon Oct 27 09:19:07 2003 From: Reuel.Cornelia <@t> tsrh.org (Reuel Cornelia) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] muscle fiber typing on goats Message-ID: Dear Friends, I just want to ask if anyone have done a muscle fiber typing (ATP'ase) on goats. If you have a procedure were i can work on it would be a great help. I have tried our procedure for human muscle and other methodology on rat muscle, it work on our 9.4 and 4.3 but did not work on our ph 4.6. I would greatly appreciate your help if anyone have tried to do a goat muscle fiber typing. thank you. Reuel Cornelia tsrh,Dallas 214-559-7699 fax: 214-559-7768 ******************************************************************************************************************* Texas Scottish Rite Hospital for Children is one of the nation's leading pediatric centers for the treatment of orthopedic conditions, certain related neurological disorders and learning differences. This email transmission and/or its attachments may contain confidential health information, intended only for the use of the individual or entity named above. The authorized recipient of this information is prohibited from disclosing it to any other party unless required to do so by law and is required to delete/destroy the information after its stated need has been fulfilled. If you are not the intended recipient, any disclosure, copying, distribution or action taken in reliance on the contents of this email transmission is prohibited. If you have received this information in error, please notify the sender immediately and delete this information. We appreciate your efforts to protect the children's confidential information. ******************************************************************************************************************* -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031027/3daf5db8/attachment.htm From rschoon <@t> email.unc.edu Mon Oct 27 09:30:30 2003 From: rschoon <@t> email.unc.edu (Robert Schoonhoven) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] RE: Digital Photomicroscopy References: <000601c39c9a$d48098c0$0200a8c0@ggrant> Message-ID: <3F9D3A16.60103@email.unc.edu> Gordon, I also have the M10AD Olympus film camera which I now compliment with the Olympus MicroFire CE (Model S99809). I say "compliment" as there is not (at this time), a true digital substitute for the resolution of a film camera. However.....I really like the MicroFire and its' software. Very easy to use and all the bells and whistles of a top of the line film camera. I don't want to sound like a commercial for Olympus but I am a very satisfied customer with this unit. Feel free to contact me off list if I can provide more information. regards, Bob Gordon Grant wrote: >Hello histonetters, >I have an Olympus BH-2 microscope fitted with a real nice PM10AD Olympus >film camera. I have been real happy with the set up as I take a lot of >publication photos. My problem is digital is now the way to go. I >borrowed a Coolpix MDC lens adapter with a coolpix and Olympus C-5050 >camera. I not very pleased with the results. Does anyone have a >recommendation for me. > >Best >Gordon Grant > > > From froyer <@t> bitstream.net Mon Oct 27 10:07:44 2003 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] (no subject) References: <123.26c65b9b.2cce8741@aol.com> Message-ID: <3F9D42D0.542EF3EB@bitstream.net> An Eosinophil is any structure or cell (most commonly a White Blood Cell) that is readily stainable with Eosin (red stain). The granules in the cytoplasm of an Eosinophil white blood cell will stain a bright red. Eosinophils increase greatly in number in certain allergic and/or parasitic diseases. Ford M. Royer, MT(ASCP) Analytical Instruments, LLC 1200 Mendelssohn Ave. N., STE 50 Minneapolis, MN 55427-4366 (800) 565-1895, Ext. 17 Fax: 952-929-1895 Cell: 612-804-1015 Email: JCarpenter764@aol.com wrote: > can anyone explain eosinophil to me From Wijaya.Martanto <@t> chbe.gatech.edu Mon Oct 27 10:13:37 2003 From: Wijaya.Martanto <@t> chbe.gatech.edu (Martanto, Wijaya) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] (no subject) Message-ID: <5D3DB84BB6F5E548AAC9FFEDC310FD091BB983@chicken-boo.che.gatech.edu> Unsubscribe please. Wijaya Martanto -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031027/ce7cbe9b/attachment.htm From mward <@t> wfubmc.edu Mon Oct 27 10:19:14 2003 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] microwave Message-ID: <61135F0455D33347B5AAE209B903A304032622E2@EXCHVS2.medctr.ad.wfubmc.edu> I am in search of a microwave oven for our lab. Our old one died on us last week. Vendors are welcome but I cannot spend over $999.00. Thanks in advance. Martha Ward Wake Forest University Baptist Medical Center -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031027/439ea4e7/attachment.htm From Linke_Noelle <@t> Allergan.com Mon Oct 27 10:27:07 2003 From: Linke_Noelle <@t> Allergan.com (Linke_Noelle) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] VEGF Message-ID: <805C4A052A253B4A8E4948B13F54D12704D407@IRMAIL102.irvine.allergan.com> Hi all! Does anyone have a decent protocol for VEGF, preferably on rat FFPE? Thank you!! Noelle Linke, BS, HTL(ASCP) Allergan, Inc 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5568 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031027/e0abeace/attachment.htm From DRG <@t> Stowers-Institute.org Mon Oct 27 12:13:59 2003 From: DRG <@t> Stowers-Institute.org (Grant, Debra) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] embedding L's Message-ID: Hello Histonetters, I have a researcher who would like to purchase some embedding L's or something very similar. If you know where I can find them please e-mail me! Thanks in advance Debby Grant Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, MO 64110 drg@stowers-institute.org From mlm11 <@t> cornell.edu Mon Oct 27 12:43:30 2003 From: mlm11 <@t> cornell.edu (Mary Lou) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] embedding L's Message-ID: <5.2.1.1.2.20031027132949.04316e20@postoffice9.mail.cornell.edu> Thermo-Shandon. here's a phone number: 800-547-7429 I don't find them in the web site but the L's are in my catalogue--5 different sizes. Cat. # 3341 thru 3345 Mary Lou Norman From kbowden <@t> ucsd.edu Mon Oct 27 13:45:50 2003 From: kbowden <@t> ucsd.edu (K. Bowden) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] RE: Digital Photomicroscopy In-Reply-To: <000601c39c9a$d48098c0$0200a8c0@ggrant> References: <000601c39c9a$d48098c0$0200a8c0@ggrant> Message-ID: <3F9D75EE.4070809@ucsd.edu> I don't have a suggestion for a camera but I have heard that for publication quality photos you need to have a 6 to 7 megapixel camera. Then once you have solved that issue you need to have the proper distant from the camera to the scope and that is the real problem. We are still working on that problem in our lab. Karen Bowden Staff Research Associate II UCSD La Jolla, CA Gordon Grant wrote: >Hello histonetters, >I have an Olympus BH-2 microscope fitted with a real nice PM10AD Olympus >film camera. I have been real happy with the set up as I take a lot of >publication photos. My problem is digital is now the way to go. I >borrowed a Coolpix MDC lens adapter with a coolpix and Olympus C-5050 >camera. I not very pleased with the results. Does anyone have a >recommendation for me. > >Best >Gordon Grant > >-----Original Message----- >From: histonet-admin@lists.utsouthwestern.edu >[mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of >histonet-request@lists.utsouthwestern.edu >Sent: Saturday, October 25, 2003 10:00 AM >To: histonet@lists.utsouthwestern.edu >Subject: Histonet digest, Vol 1 #101 - 12 msgs > >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-admin@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: Histonet digest, Vol 1 #100 - 15 msgs (amosbrooks) > 2. Re: CD 25 (Dana Settembre) > 3. Re: Moth Sperm. (Geoff McAuliffe) > 4. can someone please explain lyses (JCarpenter764@aol.com) > 5. (no subject) (Wright, Barbara (SPRI 2)) > 6. CD25 (Dawson, Glen) > 7. rhodamine auto-fluor. problem (Bonnie P Whitaker) > 8. (no subject) (JCarpenter764@aol.com) > 9. RE: (no subject) (Barry R Rittman) > 10. Reality Check (Joe Nocito) > 11. Re: CD23 and BCL-6 on B5 fixed tissue (Hadi Yaziji) > 12. RE: (no subject) (Smith, Allen) > >--__--__-- > >Message: 1 >Date: Fri, 24 Oct 2003 10:46:00 -0700 (PDT) >From: amosbrooks >Reply-To: amosbrooks@earthlink.net >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Re: Histonet digest, Vol 1 #100 - 15 msgs > >Ronnie, > We use Novocastra's CD25 (NCL CD25 305) at 1:100. We pretreat with >hot >citrate buffer (HIER) and detect with Envision. >Amos Brooks > > > >From: "Houston, Ronnie" >To: "Histonet (E-mail)" >Date: Thu, 23 Oct 2003 12:58:03 -0400 >Subject: [Histonet] CD 25 > > >Is anyone using CD25 on formalin-fixed, paraffin- >embedded human tissue? What clone and source are available? > >Thanks >Ronnie Houston >Regional Histology Operations Manager >Bon Secours HealthPartners Laboratories >5801 Bremo Road >Richmond, VA 23226 >(804) 287 7972 >ronnie_houston@bshsi.com > > >--__--__-- > >Message: 2 >Date: Fri, 24 Oct 2003 14:09:07 -0400 >From: Dana Settembre >To: Ronnie_Houston@bshsi.com, histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] CD 25 > >Hello Ronnie, >I use CD25 (Interleukin-2 Receptor) from Novocastra Laboratories which >is distributed by Vector Laboratories in Burlingame, CA. >I use in on FFPE human tissue. Their clone is 4C9. Their catalog >number is NCL-CD25-305. >Their spec sheet recommends a tonsil for a positive control. >I use it at 1:50 with Dako's Target Retrieval Solution. >Good Luck. >Dana Settembre >University Hospital-UMDNJ >Newark, NJ > > > > >>>>"Houston, Ronnie" 10/23/2003 9:58:03 AM >>>> >>>> >>>> > >Is anyone using CD25 on formalin-fixed, paraffin- >embedded human tissue? What clone and source are available? > >Thanks >Ronnie Houston >Regional Histology Operations Manager >Bon Secours HealthPartners Laboratories >5801 Bremo Road >Richmond, VA 23226 >(804) 287 7972 >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. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >--__--__-- > >Message: 3 >Date: Fri, 24 Oct 2003 14:14:46 -0400 >From: Geoff McAuliffe >To: Bruce Abaloz >Cc: HistoNet , > histonet-admin@lists.utsouthwestern.edu >Subject: Re: [Histonet] Moth Sperm. > >Can you see the nucleus? If so, I don't understand the problem. Perhaps >you are trying to stain the whole nucleated sperm one color and the >whole anculeate sperm another? Otherwise, just stain for something found > >in a nucleus and not in the cytoplasm. > >Geoff > >Bruce Abaloz wrote: > > > >>Hello Everyone, >>my name is Kathryn Mcnamara & I am a PhD student at the University of >> >> >Melbourne, Australia. I am examining the sperm of moths. Moths have 2 >types of sperm a nucleated (eupyrene) sperm and an anucleated (apyrene) >sperm. > > >>I was/AM hoping to find something that would only stain the nucleated >> >> >sperm so that the two can be easily differentiated with the one stain. >If you could help me it would be very much appreciated. THANKS in >advance, > > >>Kathryn McNamara >> >>Department of Zoology >>University of Melbourne >>Victoria, Australia 3010 >> >> >> >> >> >> > > > -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031027/346b09cb/attachment.htm From amosbrooks <@t> earthlink.net Mon Oct 27 14:12:05 2003 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Re: Histonet digest, Vol 1 #103 - 18 msgs In-Reply-To: <20031027180001.32114.96451.Mailman@swlx167.swmed.edu> References: <20031027180001.32114.96451.Mailman@swlx167.swmed.edu> Message-ID: <6.0.0.22.0.20031027150234.029a8b20@127.0.0.1> Noelle, We've been working on this for a long time with constant inconsistant results. We've used many vendors antibodies and none have performed properly. (BTW, we run a multitude of other antibodies and they all work fine so the problem is the antibody not the rest of the process) If you're doomed to follow the VEGF path I highly advise that you run negative controls on it. You're bound to notice similarities. If you read about VEGF in literature do so with a highly skeptical mind. Good luck, Amos Brooks At 01:00 PM 10/27/03, you wrote: >Hi all! > >Does anyone have a decent protocol for VEGF, preferably on rat FFPE? = > >Thank you!! > >Noelle Linke, BS, HTL(ASCP) >Allergan, Inc >2525 Dupont Drive RD-2A >Irvine, CA 92612 >714-246-5568 From northma <@t> ohsu.edu Mon Oct 27 14:27:27 2003 From: northma <@t> ohsu.edu (Mary North) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Histotech opening Message-ID: Oregon Health & Science University Portland, Oregon There is an immediate opening for a full-time histology technician, M - F, day shift, rotating on-call weekends and holdays. ASCP registry (HT or HTL), or eligible preferred. A two-year Associate Degree or equivalent experience/training in histology required. Salary range: $15.14 to 20.52 per hour. To be considered for this position, please go to our website www.ohsujobs.com, to fill out an on-line OHSU employment application, or submit a hard copy OHSU employment application to Oregon Health & Science University, Attn: HR, 3181 SW Sam Jackson Park Rd., Portland, OR 97201-3098. Please include position number IRC7156-36022P. Mary North, HT(ASCP), HTL OHSU Portland, OR -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031027/4cfad599/attachment.htm From GSLOAN <@t> mail.mcg.edu Mon Oct 27 15:58:02 2003 From: GSLOAN <@t> mail.mcg.edu (Gloria Sloan) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Citrisolv clearing Message-ID: I am using Citirsolv as a final clearing step in my staining procedure instead of xylene. How long should the clearing step take? I am leaving the slides in Citrisolv for 30 minutes since it is said to take at least twice as long to clear as xylene. The tissue is frozen mouse soleus muscle cut at 8 microns and embedded in OCT. results are good but I would like to reduce incubation times if possible. Also is Citrisolv compatible with Fisher's Permount mounting medium. Can I mount before the Citrisolv dries completely or do I need to allow it to dry? at the moment I am allowing the slides to dry overnight. Again I would like to speed up the process if I can to obtain results more quickly. Gloria Sloan Medical College of Georgia Augusta, GA 30912 From kalkonde <@t> uthscsa.edu Mon Oct 27 16:29:20 2003 From: kalkonde <@t> uthscsa.edu (Yogeshwar Kalkonde) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] mouse brains- sucrose cryoprotection- sectioning Message-ID: <3F9A3FBD@webmail.uthscsa.edu> Hi Histonetters- i am trying do Nissl staining on mice brain sections. Mice brains were fixed in 4%PFA and cryoprotected by overnight treatment with 30% sucrose. Brains were then embedded in OCT and frozen. while trying to section the the tissues in a cryostat (@8 microns thickness) the sections shrink, distorting the tissue architecture. any clue what might be going wrong? thanks in advance... yogesh Yogeshwar V. Kalkonde, MD. Post Doctoral Fellow, Department of Medicine (Infectious Diseases) University Of Texas Health Sciences Center at San Antonio, 7703 Floyd Curl Drive, 5.084R, Mail Code 7870, San Antonio, TX 78229-3900 Ph: 210-567-0385 (lab) From cwscouten <@t> myneurolab.com Mon Oct 27 16:55:36 2003 From: cwscouten <@t> myneurolab.com (Charles W. Scouten, Ph.D.) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] mouse brains- sucrose cryoprotection- sectioning Message-ID: This is a well known effect, mentioned in the atlas of Konig and Klipple (precursor to Paxinos). You didn't say, are you perfusing the brains, or just extracting them and dropping them in PFA. Definitely, you should perfuse for better quality, to get fixative everywhere fast. Either way, you will get shrinkage and distortion unless you perfuse with a sucrose prewash at 300 mmHg, followed by the fixative. An apparatus is available, a protocol is posted, and the rational is given at the following link. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=471001&catdesc=Histology+Equipment&CatThreeID=674&CatOneID=4&subcatdesc=Sacrifice+Equipment&idsubcategory=21 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 www.myneurolab.com -----Original Message----- From: Yogeshwar Kalkonde [mailto:kalkonde@uthscsa.edu] Sent: Monday, October 27, 2003 4:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] mouse brains- sucrose cryoprotection- sectioning Hi Histonetters- i am trying do Nissl staining on mice brain sections. Mice brains were fixed in 4%PFA and cryoprotected by overnight treatment with 30% sucrose. Brains were then embedded in OCT and frozen. while trying to section the the tissues in a cryostat (@8 microns thickness) the sections shrink, distorting the tissue architecture. any clue what might be going wrong? thanks in advance... yogesh Yogeshwar V. Kalkonde, MD. Post Doctoral Fellow, Department of Medicine (Infectious Diseases) University Of Texas Health Sciences Center at San Antonio, 7703 Floyd Curl Drive, 5.084R, Mail Code 7870, San Antonio, TX 78229-3900 Ph: 210-567-0385 (lab) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From edonato <@t> fhcrc.org Mon Oct 27 17:55:18 2003 From: edonato <@t> fhcrc.org (Donato, Elizabeth M) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] help Message-ID: <07470B0DBFBDD511BD770002B330A37DC39F2C@curly.fhcrc.org> -----Original Message----- From: HistoNet Server [mailto:histonet@pathology.swmed.edu] Sent: Wednesday, December 01, 1999 7:31 PM To: Donato, Elizabeth M Subject: re: subscribe digest Your address has been added to the addresses that comprise this Digest List. Welcome to HISTONET. This is an electronic mailing list for the exchange of information pertaining to histotechnology and related fields. PLEASE SAVE THIS MESSAGE. It contains useful information about how to use the list and what to do if you experience problems. It also includes some basic rules for email etiquette (Netiquette) which will be helpful to those who are new to this form of communication. WHAT IS A LISTSERVER? A list server is a computer that runs software which will receive incoming electronic mail (email) messages and reroute them automatically to everyone on the subscriber list. Email uses the vast expanse of the Internet to allow almost instantaneous communication between networked computers around the world. Our system uses the LISTSTAR software from Quarterdeck Corporation (California) and can currently send about 30 messages a minute. With the present number of subscribers, we are processing about 10,000 outbound messages a day. WHO SHOULD SUBSCRIBE? Anyone interested in research or clinical applications of histology, immunohistochemistry, in-situ hybridization pathology, and electron microscopy may find Histonet informative and useful. Currently, there are more than 850 subscribers from all over the world. Subscribers include hospital employees from major urban centers and obscure remote locales, university researchers, botanists and the employees of commercial laboratories, government agencies, veterinary facilities and a wide variety of commercial industrial ventures. WHO RUNS HISTONET? The list is run by Linda R. Margraf, M.D. and Herb K. Hagler, Ph.D. using hardware and software owned by the University of Texas Southwestern Medical School, Department of Pathology in Dallas, Texas. If you have any questions or problems with Histonet please contact Linda Margraf at LMargraf@childmed.dallas.tx.us. HOW DOES THE LIST WORK? This server, unlike many systems, uses ONLY ONE ADDRESS to send commands to the computer and to post messages. The server will recognize commands sent in the SUBJECT line of the message and only when they are spelled exactly as listed below. Anything not identified as a command will be circulated to EVERYONE on the list. The following is a list of commands the server recognizes: subscribe Your address will be added to the list of subscribers. You will then be able to send messages to this list that will be forwarded to all other list subscribers. You will begin to receive all messages sent to the list by other subscribers. subscribe digest Your address will be added to the list of subscribers who receive a digest instead of each forwarded message. A digest is a compilation of all the messages received in a 24 hour period. It is sent to the digest subscribers every night after midnight. Digest subscribers can post and respond to messages the same as "real-time" subscribers. digests A list of available digests will be returned to you. Histonet stores old messages as daily digests for approximately three months. To read previous messages, copy the list of available digests, mark the dates of interest and return it to the server. unsubscribe Your address will be removed from the list of subscribers. You will no longer be able to send messages to the members of the list. help A list of the commands recognized by the server will be returned to you. WHAT ARE THE RULES? You may post any questions you wish pertaining to histology, pathology, in-situ hybridization, immunohistochemistry etc. Equipment and reagent evaluations, laboratory management issues, government regulations, and job opportunities are all appropriate topics. The University asks that we restrict the use of its hardware and software to business purposes only (occasional jokes do slip through but PLEASE use restraint). Vendors and those with commercial interests in histology products are welcome contributors however, we ask that blatant advertisements be avoided at all times. It is fine to refer to product that your company produces if it is pertinent to a topic being discussed on the list. Unsolicited advertisements are poorly tolerated by the members and you will likely receive a number of negative comments if you overstep the boundaries. Please contact Linda Margraf at LMargraf@childmed.dallas.tx.us if you are not sure about the appropriateness if a message you wish to post. BASIC HISTONET "NETIQUETTE" It is most helpful to the list members if you post your responses to queries to everyone on the list and not just as a personal reply to the person asking the question. That way duplicate messages are minimized and we all learn from each other's comments. Likewise, if you post a question and get a number of responses back directly to you, it is helpful to everyone if you could send out a summary of the replies you got to Histonet. Please avoid abbreviations unless they are explained in your message. For example: immunohistochemistry (IHC). This list circulates to a wide variety of individuals and what seems obvious to you may have no meaning on the other side of the world. Please sign your letter and include your institution or affiliation and location. Not all email systems have headers which identify the sender. Do use the subject line to indicate the topic of your message. DON'T USE ONLY CAPITAL LETTERS -it is considered shouting. Please send questions and problems about the list directly to Linda Margraf at LMargraf@childmed.dallas.tx.us and don't circulate them to the >850 subscribers on the list. Be careful when sending commands to the server to put the command in the SUBJECT LINE and spell it correctly. Please do not send images as attachments with your message. We can now post images at our web site (http://pathcuri1.swmed.edu). To have an image posted send it to Herb Hagler at herb.hagler@email.swmed.edu. es From Sellis4051 <@t> aol.com Mon Oct 27 18:09:11 2003 From: Sellis4051 <@t> aol.com (Sellis4051@aol.com) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Histology's contribution to Forensic Science Message-ID: <186.2101f450.2ccf0da7@aol.com> Karen, it sounds like you are on the same path that I took to working in forensics. I loved working in the morgue as a college student; I was a Chinese major then. From there I got referrals to other morgues in Seattle and worked part time at a couple different places. I agree with you that there is always something new to learn at every autopsy, maybe more so at hospital autopsies where multiple chronic conditions can complicate the clinical picture. Unfortunately, as you mentioned, hospital autopsy numbers are way down because no one can figure out how to charge for them. Ideally, funding would come through the hospital's Total Quality Management system, rather than being seen as an expense to the pathology department. You mention time and money as the hindrance to entering the field of forensics. In my 20's without children, my biggest asset was my low financial expectations and seemingly limitless energy. Working for free or being willing to led to all my big breaks in the quest for forensic employment. I highly recommend it. Also some creative thinking can lead most anything into something forensically relevant. I volunteered at a hospice so that I could get comfortable with grieving people. The Masters that I'm getting in Forensic Science Administration should help even out my education and if I continue to be a full time student, I'll be done 12/04. But I have a lead on a job with my home town coroner, which may slow my studies down. Wish me luck! Sandra Ellis HT (ASCP) In a message dated 10/21/2003 6:03:48 AM Pacific Daylight Time, Bauer.Karen@mayo.edu writes: This sounds very interesting to me. I am a Histology Supervisor who is also the Hospital Deaner. The Pathologists trained me to be the Prosector for autopsies and I'm one of those strange people who enjoy it. I find the human body very interesting and every autopsy is a learning experience. I work at a hospital that only does autopsies on inpatients that have passed away, and that's only 3 for the year so far. I've always thought about going into Forensics, or at least, learning more about it. Time and money usually prevent me from doing so. I congratulate you on getting your Masters! Good luck in your search. Karen Bauer, HT(ASCP) -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031027/6da034c0/attachment.htm From Sellis4051 <@t> aol.com Mon Oct 27 18:20:41 2003 From: Sellis4051 <@t> aol.com (Sellis4051@aol.com) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Histology's contribution to Forensic Science Message-ID: <18a.2134698e.2ccf1059@aol.com> Thanks for your response. After reading your email, I decided that I would have a part of my presentation focus on the detriments due to the poor handling that these medical examiners give their tissues. So I'll be including some mention of the consequences of improper fixation, including your IHC concern. I'm still hoping that someone out there will have had an experience that firmly links histology to the proper resolution of a death investigation case. Sandra Ellis In a message dated 10/21/2003 7:33:50 AM Pacific Daylight Time, LBlack@carilion.com writes: Sandra, The Histology Lab where I work processes ME cases. I believe the specimens are sometimes held at the ME's office for several days/weeks before they are sent to us. We perform these cases as we have time. The specimens still have to be trimmed in to an appropriate size before placing into cassettes. My concern is when immunoperoxidase stains are requested on these cases after a prolonged stay in fixative, a false negative stain may result due to masking of antigenic determinates. For hospital surgical cases, we always have the tissues processed the same day if possible. Even with a weekend holiday, specimens are not held in fixative for longer than 72 hours. Sounds like an interesting job you have. Best wishes. Lisa Black Carilion Consolidated Laboratory -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031027/6ef7c50c/attachment.htm From katri <@t> cogeco.ca Mon Oct 27 18:29:01 2003 From: katri <@t> cogeco.ca ( Katri Tuomala) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Explain the biochemical basis of 'no counterstain with hematoxylin' References: <200310261957.h9QJv0wv008505@korotoa.bdonline.com> Message-ID: <001901c39cea$7e985a60$ce989618@hala4.on.cogeco.ca> Subratab, You are using very harsh retrieval for something that should not need any at all. Methacarn fixative is not crosslinking, therefore you are destroying your tissue by proteolytic enzymes and heat. Even tissue that has been fixed with formalin, but not long enough, produces diminished nuclear staining. Try your method without any retrieval, when fixed in methacarn. Katri Katri Tuomala St.Joseph's Healthcare Hamilton, Ontario, Canada ----- Original Message ----- From: "Subratab" To: Sent: Sunday, October 26, 2003 2:53 PM Subject: [Histonet] Explain the biochemical basis of 'no counterstain with hematoxylin' > Dear all > I am staining (IHC) rat renal tissue for ED1. When I am using > Methacarn-fixed tissue I am not getting any counterstain with hematoxylin. > Nuclear positions are showing blank space (ghost-like rounded gaps). Tissue > is showing just-yellowish appearance. > I have stained the methacarn-fixed slides with hematoxylin without going > through my IHC protocol (deparaffinization and hematoxylin). And the > staining is OK. So the methacarn-fixed tissue has no problem with > hematoxylin. It indicates that my IHC protocol is not campatible with > hematoxylin stain when I am using methacarn-fixed tissue. > > On the otherhand, when I am using paraformaldehyde fixed tissue with the > same protocol, I am facing no problem to counterstain with hematoxylin. > 1. Can you please explain the biochemical basis? 2. Can the problem be > solved by methyl green counterstaining? > I am expecting clarification and suggestion from the histonet > experts.Following is my IHC protocol: > > Deparaffinization and rehydration > > Digestion of protein cross-links by trypsin > > Microwave exposure in citrate buffer > > Blocking with 1% non-fat milk > > Primary Ab (ED1) diluted in PBS (1% BSA + 0.3% Triton x100) > > 3% H2O2 in methanol > > AP conjugated polymer (anti-mouse Ig) > > Fast red with substrate buffer and levamisol (from Dako) > > Counterstaining with hematoxylin and mounting with permafluor. > > Washing solution is PBS. (ED1 staining is OK. The problem is with > counterstaining) > > Subrata Biswas. > University of Campinas, brazil. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From doscwk <@t> nus.edu.sg Mon Oct 27 18:43:35 2003 From: doscwk <@t> nus.edu.sg (Chan Wai Kam) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] embedding L's Message-ID: Hi Debby, I bought mine from Leica. Julee Chan Orthopaedic Surgery National University of Singapore -----Original Message----- From: Grant, Debra [mailto:DRG@Stowers-Institute.org] Sent: Tue 10/28/2003 2:13 AM To: Histonet (E-mail) Cc: Subject: [Histonet] embedding L's Hello Histonetters, I have a researcher who would like to purchase some embedding L's or something very similar. If you know where I can find them please e-mail me! Thanks in advance Debby Grant Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, MO 64110 drg@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From katri <@t> cogeco.ca Mon Oct 27 18:47:42 2003 From: katri <@t> cogeco.ca ( Katri Tuomala) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Explain the biochemical basis of 'no counterstain with hematoxylin' References: <200310261957.h9QJv0wv008505@korotoa.bdonline.com> Message-ID: <001d01c39ced$18a737a0$ce989618@hala4.on.cogeco.ca> One more thing I want to add to my previous message, Subratab. In your IHC protocol, you use alkaline phosphatase conjugated secondary antibody. You don't need the 3% H2O2 in methanol step, which is to block endogenous peroxidase. That is also too harsh for alcohol fixed tissue. Katri ----- Original Message ----- From: "Subratab" To: Sent: Sunday, October 26, 2003 2:53 PM Subject: [Histonet] Explain the biochemical basis of 'no counterstain with hematoxylin' > Dear all > I am staining (IHC) rat renal tissue for ED1. When I am using > Methacarn-fixed tissue I am not getting any counterstain with hematoxylin. > Nuclear positions are showing blank space (ghost-like rounded gaps). Tissue > is showing just-yellowish appearance. > I have stained the methacarn-fixed slides with hematoxylin without going > through my IHC protocol (deparaffinization and hematoxylin). And the > staining is OK. So the methacarn-fixed tissue has no problem with > hematoxylin. It indicates that my IHC protocol is not campatible with > hematoxylin stain when I am using methacarn-fixed tissue. > > On the otherhand, when I am using paraformaldehyde fixed tissue with the > same protocol, I am facing no problem to counterstain with hematoxylin. > 1. Can you please explain the biochemical basis? 2. Can the problem be > solved by methyl green counterstaining? > I am expecting clarification and suggestion from the histonet > experts.Following is my IHC protocol: > > Deparaffinization and rehydration > > Digestion of protein cross-links by trypsin > > Microwave exposure in citrate buffer > > Blocking with 1% non-fat milk > > Primary Ab (ED1) diluted in PBS (1% BSA + 0.3% Triton x100) > > 3% H2O2 in methanol > > AP conjugated polymer (anti-mouse Ig) > > Fast red with substrate buffer and levamisol (from Dako) > > Counterstaining with hematoxylin and mounting with permafluor. > > Washing solution is PBS. (ED1 staining is OK. The problem is with > counterstaining) > > Subrata Biswas. > University of Campinas, brazil. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kutaisi13 <@t> hotmail.com Mon Oct 27 19:38:08 2003 From: kutaisi13 <@t> hotmail.com (david moses) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Help with cell culture Message-ID: Dear Histonetters I'd highly appreciate your opinion on following issue: Neural cell cultures are fixed with 4% PFA and then permeabilized with Triton.Some people use brief exposure to Triton some incubate primary Ab for quite a long time with Triton. Another approach is to fix/permeate cultures with methanol 20 min for -20. I wondered about the impact of different methods on antigen detection - tubulin,NF,GFAP etc'.Of course,excluding oligodendrocyte stains. Thanks David Moses Research Fellow _________________________________________________________________ Protect your PC - get McAfee.com VirusScan Online http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From katri <@t> cogeco.ca Mon Oct 27 19:42:40 2003 From: katri <@t> cogeco.ca ( Katri Tuomala) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Anyone have a favorite CD15 clone? Message-ID: <000a01c39cf4$c6840540$ce989618@hala4.on.cogeco.ca> ----- Original Message ----- From: " Katri Tuomala" To: "Drew Sally A." ; "Histonet" Sent: Monday, October 27, 2003 8:15 PM Subject: Re: [Histonet] Anyone have a favorite CD15 clone? > Hi Sally, > > I'm not in the lab at the moment, but off top of my head I remember reading, > that LeuM1 antibody only marks a certain percentage of the RC cells and > threfore we always also do CD30 in suspected Hodgkins cases. > > Katri > > Katri Tuomala > St Joseph's Healtcare > Hamilton, Ontario, Canada > > > ----- Original Message ----- > From: "Drew Sally A." > To: "Histonet" > Sent: Tuesday, October 21, 2003 3:39 PM > Subject: [Histonet] Anyone have a favorite CD15 clone? > > > We are just wondering if not all clones of CD15 are equal, and that maybe > someone > out there has tested more than a couple different CD15's? We have > pathologists > who periodically ask us to repeat our stain because they expect to see more > RS cells staining than they do. We currently run our antibodies on Ventana > immunostainers if anyone feels that makes a difference. > All constructive opinions are welcome. > Thank you! > > Sally Ann Drew-MT(ASCP) > Univ. of Wisconsin Hospital & Clinics > IHC/ISH Laboratory > 600 Highland Ave. VAH-DM223 > Madison, WI 53792-2472 > 608-265-6596 > sa.drew@hosp.wisc.edu > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From lpwenk <@t> covad.net Mon Oct 27 20:04:06 2003 From: lpwenk <@t> covad.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] (no subject) References: <71.369a1b95.2cce8696@aol.com> Message-ID: <005301c39cf7$c4d11dc0$8732fea9@hppav> An anhydrous chemical has NO water attached to it. A trihydrate chemical has 3 waters attached to it. This becomes important when making solutions, as the weight of the water has to be included in the weight of the compound. 100 g of sodium acetate, trihydrate is NOT the same amount of sodium acetate as 100 g of sodium acetate, anhydrous. Some of the weight of the sodium acetate trihydrate is the water. The molecular weight of sodium acetate anhydrous is about 82. (CH3COONa) = (2 Carbons x 12) + (3 Hydrogens x 1) + (2 Oxygens x 16) + (1 Sodium x 23) (You need to look at a periodic table to get the weights of the chemicals, and the side of the container to get the formula. Often, the side of the container also lists the molecular weight.) The molecular weight of sodium acetate trihydrate is about 136. (CH3COONa*3H2O) = weight of the anhydrous (82) plus the weight of 6 more hydrogens (+6) plus 3 more oxygens (+48) So, if the directions calls for 100 g of the anhydrous sodium acetate, but you only have trihydrate sodium acetate, you need to set up a ratio: x = (136/82) x 100 = 165.8 g This means that 165.8 g of sodium acetate trihydrate to equal 100 g of sodium acetate anhydrous. So to make up the solution, you would use the 165.8 g of sodium acetate trihydrate, instead of the 100 g. Hope this helps. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: JCarpenter764@aol.com To: histonet@lists.utsouthwestern.edu Sent: Monday, October 27, 2003 9:32 AM Subject: [Histonet] (no subject) what is the difference between the anhydrous compound and the trihydrate compound? -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031027/a0eeda2e/attachment.htm From mikael.niku <@t> helsinki.fi Tue Oct 28 01:57:19 2003 From: mikael.niku <@t> helsinki.fi (Mikael Niku) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] RE: Digital Photomicroscopy In-Reply-To: <000601c39c9a$d48098c0$0200a8c0@ggrant> Message-ID: <000501c39d29$1cc35850$8c0fd680@ekk1116> Gordon, we have a BH-2 and changed from film to digital a few ears ago. We first tried a few "normal" digital cameras the Nikon D1 digital SLR, for example), but it seemed o be difficult to attach the camera to the scope properly you probably need an expert adaptor company to do his, at least with the old scopes). With the cameras specifically designed for microscope work there were no major problems, and we finally got a Colorview12 camera + image analysis software by Soft Imaging System. With a resolution of 1280x1024 pixels and less than perfect color reproduction, there are better cameras available today. But even this IS enough to produce decent publication images (the minimum resolution most journals ask is 150-200 dpi, so you can produce images at least 16 cm wide, which is of course much larger than most images in scientific publications). I actually won a honorable mention in the Nikon international photomicrography competition last year using this camera, so it can't be that bad. I never used film since getting the digital camera; the lot of slide film I bought slightly before is still sitting in the freezer. It is true that the resolution of even the best digital cameras is theoretically less than that of film. But for most scientific publishing, there is no practical difference (see above). If I understand correctly, the resolution of modern digital cameras is actually so high that at least with moderate to high magnification the limiting factor is the resolution of the microscope objective, not the camera. So you don't really get any extra information using film. My view is that with digital equipment one can produce at least as good quality images as with film, but digital is a LOT easier and quicker to use. You must note, though, that to get really optimal results, you need to spend some time learning digital color reproduction and image processing. But this is no different with film. +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ + Mikael Niku + University of Helsinki, Dept. Basic Veterinary Sciences + URL: www.helsinki.fi/~mniku/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! (Gandhi) > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of > Gordon Grant > Sent: 27. lokakuuta 2003 16:59 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] RE: Digital Photomicroscopy > > > Hello histonetters, > I have an Olympus BH-2 microscope fitted with a real nice > PM10AD Olympus film camera. I have been real happy with the > set up as I take a lot of publication photos. My problem is > digital is now the way to go. I borrowed a Coolpix MDC lens > adapter with a coolpix and Olympus C-5050 camera. I not very > pleased with the results. Does anyone have a recommendation for me. > > Best > Gordon Grant From louise_renton <@t> hotmail.com Tue Oct 28 02:43:22 2003 From: louise_renton <@t> hotmail.com (louise renton) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] (no subject) Message-ID: Firstly, I would recommend that you invest in a good, preferably a scientific dictionary. This will stand you in good stead throughout your career, even when there is a power failure and the internet is down. Secondly, I will voice my suspicion that you are "yanking" our collective chain. You make no mention of the institution to whom you are affiliated, or indeed what it is that you are doing. Whereas the the knowledge we have is to be freely shared, some of us neither have the time nor the inclination to sift the genuine queries from the false. Louise Renton Bone Research Unit MRC Johannesburg South Africa Tel & fax +27 11 717 2298 "Time flies like an arrow, fruit flies like a banana" ----Original Message Follows---- From: JCarpenter764@aol.com To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Date: Mon, 27 Oct 2003 09:32:54 EST what is the difference between the anhydrous compound and the trihydrate compound? _________________________________________________________________ Get your own, free Web space - on MSN Groups! http://groups.msn.com/people?pgmarket=en-za From ree3 <@t> leicester.ac.uk Tue Oct 28 04:14:44 2003 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] GST omega 1 Message-ID: Looking for a supplier for an antibody to the above..............many thanks.. Richard Edwards MRC TOXICOLOGY UNIT... LEICESTER....U.K....... From mike.kirby <@t> nhls.ac.za Tue Oct 28 04:28:40 2003 From: mike.kirby <@t> nhls.ac.za (Mike Kirby) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Neutralisation of Formaldehyde using Monoethanolamine solution Message-ID: <0FA1B7472216894D9B3381F87C71DE491DA43B@nhlsmail.nhls.ac.za> Dear Netters. Recently, I read an article which described the neutralisation of formaldehyde in cadavers by infusing them with a 4% solution of Monoethanolamine, and I was wondering if the same system could be used for disposing of used formalin in the Anatomical Path Lab. Has anyone got anything to offer on this subject? Mr.M.Kirby. Chief Safety Officer National Health Lab Service Johannesburg South Africa. From deirdre_brophy <@t> hotmail.com Tue Oct 28 06:42:45 2003 From: deirdre_brophy <@t> hotmail.com (Deirdre Brophy) Date: Fri Sep 16 15:22:05 2005 Subject: [Histonet] Help with frozen sections Message-ID: An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/81730d66/attachment.htm From abright <@t> brightinstruments.com Tue Oct 28 08:29:06 2003 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Help with frozen sections Message-ID: Dear Deirdre, You do not state the temperature of the tissue you are trying to section in the cryostat. I think that raising the temperature would eliminate the cracking. Best Regards Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: Deirdre Brophy [mailto:deirdre_brophy@hotmail.com] Sent: 28 October 2003 12:43 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Help with frozen sections Hello, Does anyone have any advice on getting good frozen sections from tissue that has been stored in formaldehyde for prolonged periods? I am currently cutting fish gonads on a cryostat. This could potentially save a lot of time and expense compared to wax histology. My samples have been stored in 4% buffered formaldehyde for several months. Due to the nature of my sampling, this is difficult to avoid. I have been cryoprotecing the tissue with a mixture of DMSO and sucrose, and then freezing in a bath of hexane cooled with dry ice. I am taking 6-10um sections which are then stained with H&E and viewed with a light microscope. For the female gonads, this is working quite well, the morphology is good and cryprotection seems to keep freeze damage to a minimum. However, for the testes the results are not so good. Sections are cracked and morphology is really poor. When the same tissue is cut using wax histology the sections are fine. Rinsing the tissue well in PBS to remove excess formaldehyde before sectionning on the cryostat helps, as does cryprotecting for longer periods (overnight) but cracks are s till visible. Could prolonged storage in formaldehyde render the tissue less permeable to the cryoprotectant perhaps? Any suggestions/remedies would be much appreciated as I am running out of things to try! Best wishes, Deirdre _______________________ Deirdre Brophy Dept of Life Sciences GMIT Dublin rd Galway Ireland _____ MSN 8 helps ELIMINATE E-MAIL VIRUSES. Get 2 months FREE*. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From aep14 <@t> leicester.ac.uk Tue Oct 28 09:31:13 2003 From: aep14 <@t> leicester.ac.uk (Prior, A.E.) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Chitin Softening- Results Message-ID: <6728E9270FDF8D44A41AF135DBB553E846D4A9@sumac.cfs.le.ac.uk> A few months ago I wrote asking for help regarding sectioning the eyes from Edible Crabs. Many thanks for all the suggestions. In between my proper work I managed to test a few of these out, and these are the results in brief: All eyes were collected fresh and fixed for at least 48 hours in formal saline (10%).Dehydrations were done using propan-2-ol rather than ethanol to reduce hardening effects. Embedded in Paramat wax via Histoclear. The shell of the crab is a mix of calcium and chitin. Eyes were placed in 10% formic acid (18-24 hours with 2 changes)under vacuum, until soft and all calium removed. Eyes were then treated with either sodium hydoxide, phenol/chloral hydrate mix or Nair ( a commercial hair removal cream available from all good chemists (that's a drug store to all you Americans)) NaOH (24 h in 2% soln.)treatment softened the chitin enough so sections could be cut with extreme care. The phenol treatment was less successful. Even after a week in the mix the chitin was still too hard to section properly. Tissue preservation was poor in these samples, but unsure whether this was solely due to the phenol. Coating the eyes in Nair for 24 hours produced eyes that section well. Slower speeds than normal were needed to avoid thickness variation in sections, but was quicker than the NaOH treated eyes. Tissue preservation was excellent as the Nair did not penetrate the eyes. Thanks again for all the help on this. Hope these results are useful to people - I have the protocols if people need further details, contact me directly. Andrew Prior Biology Dept. Leicester University UK aep14@le.ac.uk From srrx2 <@t> netzero.net Tue Oct 28 09:40:09 2003 From: srrx2 <@t> netzero.net (srrx2@netzero.net) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] florida techs Message-ID: <20031028.074103.19995.121705@webmail09.nyc.untd.com> what is the criteria for working as a histotech in Florida...also can someone tell me how to go about licensure in the state of Florida? thanks in advance! renee From JCarpenter764 <@t> aol.com Tue Oct 28 09:45:14 2003 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] (no subject) Message-ID: <77.1b7bcccf.2ccfe90a@aol.com> while studying for my exam on the different fixatives and there ingredients....i have noticed that some call for distilled water and some use the term deionized water. Is there a difference? -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/26bcc3ad/attachment.htm From kgrobert <@t> rci.rutgers.edu Tue Oct 28 09:51:07 2003 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Grocott's stain for fungi Message-ID: <3F9E906B.7060009@rci.rutgers.edu> Hi all- I'm making up the various stock & working solutions for Grocott's stain, and I am having a heck of a time getting the 3% methenamine to go into solution. (Not methenamine silver, just plain old 3% methenamine in distilled water.) I know that you have to acid-clean the glassware for the methenamine silver & silver nitrate solutions, but do I have to do it for 3% methenamine? I tried heating it, but that did not make a bit of difference. AFIP manual has no other instructions other than "3 g methenamine in 100 ml distilled water". Is there a shelf life for methenamine? The bottle we have is pretty old, but I am not sure if that would be a factor. I intend to try acid-cleaning a bottle for the 3% methenamine this afternoon, but I wanted to see if anybody had any other suggestions to try as well. Thanks in advance- Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Dept of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers University From pmarcum <@t> polysciences.com Tue Oct 28 10:04:38 2003 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] (no subject) In-Reply-To: <77.1b7bcccf.2ccfe90a@aol.com> Message-ID: <001d01c39d6d$30468570$4800a8c0@PMARCUM2K> Please use a subject line in e-mails and also where are you working from. Many of your questions are extremely basic and should have been answered in test or training. I am sorry we have had such a flurry of e-mails with no subject line that are garbage or junk e-mail list so at this point a subject line is becoming necessary if you want an answer. Pam Marcum -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of JCarpenter764@aol.com Sent: Tuesday, October 28, 2003 10:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) while studying for my exam on the different fixatives and there ingredients....i have noticed that some call for distilled water and some use the term deionized water. Is there a difference? -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/131bb468/attachment.htm From JCarpenter764 <@t> aol.com Tue Oct 28 10:26:34 2003 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] RE: PAM MARCUM Message-ID: OK.....i understand about the subject lines. But i have been studying and working very hard at preparing myself for the exam this summer....I do appreciate every little bit of help that i have received and don't take any of it for granted. There are some basic questions that confuse me and i thought it was ok to ask for help. I don't consider this garbage, nor do i consider this junk email. Thank you for your time -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/517bb1a4/attachment.htm From thallada <@t> noch.org Tue Oct 28 10:42:44 2003 From: thallada <@t> noch.org (Hallada, Teri) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] RE: Microwave Tissue Processors Message-ID: I was wondering if anyone out there is using a microwave tissue processor for routine hospital tissues. Are there any regulations applicable to instituting one, ie FDA approval? Teri Hallada BS MT CT (ASCP) thallada@noch.org From Tony.Fearn <@t> cd.burnleyhc-tr.nwest.nhs.uk Tue Oct 28 10:44:32 2003 From: Tony.Fearn <@t> cd.burnleyhc-tr.nwest.nhs.uk (Fearn Tony) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Genomics, Proteomics and Transcriptomics Message-ID: <8C3DE9069A06D611B8950002A550C15943BCCE@BHC_MAIL02> Hello everyone, I have a Trainee Biomedical Scientist who has to complete an assignment on assessing the relative importance and significance of genomics, proteomics and transcriptomics for her stage 3 BSc in Biomedical Science. The books we have in the lab do not cover these -omics and we are struggling to find information on the internet. I wonder if there is anyone out there that could point us to books or websites that will introduce us to the BASIC outlines. Catherine has asked that it shouldn't be too high-tech. Any help would be appreciated. Best wishes Tony Fearn Anthony J Fearn Chief Biomedical Scientist Histopathology Burnley General Hospital Burnley BB10 2PQ Lancs, UK From Sven.Terclavers <@t> med.kuleuven.ac.be Tue Oct 28 10:54:40 2003 From: Sven.Terclavers <@t> med.kuleuven.ac.be (Sven Terclavers) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Looking for old article Message-ID: <002f01c39d74$2dddbea0$85f53a86@microscopy1> Hi all, I'm looking for a pdf or word-document of the following qrticle. Can anyone help me out? Thanks! Histochemistry. 1985;82(3):205-8. Cowen T, Haven AJ, Burnstock G. Pontamine sky blue: a counterstain for background autofluorescence in fluorescence and immunofluorescence histochemistry. Sven Terclavers From pmarcum <@t> polysciences.com Tue Oct 28 10:44:59 2003 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] RE: PAM MARCUM In-Reply-To: Message-ID: <002901c39d72$d34b8400$4800a8c0@PMARCUM2K> I think you misunderstood we get them (junk e-mails ans spam) from many sources and they are primarily from e-mails with no subject line that look official. I have no problem answering your questions or helping. The reference to garbage and junk mail was not about your questions just repeated failure to use a subject line heading. It is a simple request and after deleting a pile of e-mails this morning from you and several others I made a request that is only etiquette. As to your questions they have been very simple and should be easy to find in a library or good histology test. If you do not have these available and require help anyone on Histonet will attempt to help you. We can also suggest texts that may assist with your studies if you wish. Pam Marcum -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of JCarpenter764@aol.com Sent: Tuesday, October 28, 2003 11:27 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: PAM MARCUM OK.....i understand about the subject lines. But i have been studying and working very hard at preparing myself for the exam this summer....I do appreciate every little bit of help that i have received and don't take any of it for granted. There are some basic questions that confuse me and i thought it was ok to ask for help. I don't consider this garbage, nor do i consider this junk email. Thank you for your time -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/3733c8ad/attachment.htm From Jackie.O'Connor <@t> abbott.com Tue Oct 28 10:53:30 2003 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] RE: PAM MARCUM Message-ID: I agree. Just because the questions "should have been" answered somewhere in training, apparently, they weren't. Where else can people turn to for answers when they are trying to learn. I don't consider this junk or garbage mail - there are no stupid questions except the ones that are not asked. Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories GPRD JCarpenter764@aol.com Sent by: histonet-admin@lists.utsouthwestern.edu 10/28/2003 10:26 AM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] RE: PAM MARCUM OK.....i understand about the subject lines. But i have been studying and working very hard at preparing myself for the exam this summer....I do appreciate every little bit of help that i have received and don't take any of it for granted. There are some basic questions that confuse me and i thought it was ok to ask for help. I don't consider this garbage, nor do i consider this junk email. Thank you for your time -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/2e8b3fdd/attachment.htm From MGomez <@t> ameripath.com Tue Oct 28 11:01:42 2003 From: MGomez <@t> ameripath.com (MGomez@ameripath.com) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Microwave Tissue Processors Message-ID: There is an interesting article in the October issue of Cap Today for those of you who are interested in microwave processing. You may also contact Hacker Instruments at Hackerlab@aol.com or 973-226-8450 for info on their products. Regards, Milton (801) 256-0040 x254 From Jackie.O'Connor <@t> abbott.com Tue Oct 28 11:01:45 2003 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Grocott's stain for fungi Message-ID: Make sure you are using the correct chemical, it should be Hexamethylenetetramine (methenamine) (C6H12N4). It should dissolve readily in H20. Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics Kathleen Roberts Sent by: histonet-admin@lists.utsouthwestern.edu 10/28/2003 09:51 AM To: Histonet cc: Subject: [Histonet] Grocott's stain for fungi Hi all- I'm making up the various stock & working solutions for Grocott's stain, and I am having a heck of a time getting the 3% methenamine to go into solution. (Not methenamine silver, just plain old 3% methenamine in distilled water.) I know that you have to acid-clean the glassware for the methenamine silver & silver nitrate solutions, but do I have to do it for 3% methenamine? I tried heating it, but that did not make a bit of difference. AFIP manual has no other instructions other than "3 g methenamine in 100 ml distilled water". Is there a shelf life for methenamine? The bottle we have is pretty old, but I am not sure if that would be a factor. I intend to try acid-cleaning a bottle for the 3% methenamine this afternoon, but I wanted to see if anybody had any other suggestions to try as well. Thanks in advance- Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Dept of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/2257312b/attachment.htm From Salvacion.DeLovino <@t> cshs.org Tue Oct 28 11:06:35 2003 From: Salvacion.DeLovino <@t> cshs.org (DeLovino, Salvacion S.) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Grocott's stain for fungi Message-ID: Preparing 3% methenamine is straightforward. No heat needed. It melts right away in DI water. Salvacion S. Delovino -----Original Message----- From: Kathleen Roberts [mailto:kgrobert@rci.rutgers.edu] Sent: Tuesday, October 28, 2003 7:51 AM To: Histonet Subject: [Histonet] Grocott's stain for fungi Hi all- I'm making up the various stock & working solutions for Grocott's stain, and I am having a heck of a time getting the 3% methenamine to go into solution. (Not methenamine silver, just plain old 3% methenamine in distilled water.) I know that you have to acid-clean the glassware for the methenamine silver & silver nitrate solutions, but do I have to do it for 3% methenamine? I tried heating it, but that did not make a bit of difference. AFIP manual has no other instructions other than "3 g methenamine in 100 ml distilled water". Is there a shelf life for methenamine? The bottle we have is pretty old, but I am not sure if that would be a factor. I intend to try acid-cleaning a bottle for the 3% methenamine this afternoon, but I wanted to see if anybody had any other suggestions to try as well. Thanks in advance- Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Dept of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Tue Oct 28 11:08:02 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] RE: Digital Photomicroscopy Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CC68@usca0082k03.rallansci.apogent.com> Mikael writes: << If I understand correctly, the resolution of modern digital cameras is actually so high that at least with moderate to high magnification the limiting factor is the resolution of the microscope objective, not the camera. So you don't really get any extra information using film.>> That is true. It may seem backwards to some, but the higher the objective magnification, the less camera resolution needed. That is because you are imaging a very small field of view. Even a 1 or 2 megapixel camera can get good images with a 100x oil objective. You would only need very high megapixels if you were trying to get maximum resolution at very low magnification - say 4x. Since that is the case, the limiting factor becomes the quality of the optics. We saw that here when we demoed a high-end Nikon 40X oil lens and compared it against our standard dry 40X. The difference was amazing. But you'll spend $4000 for that lens, so you have to decide on the trade offs. I would say for 20x objective and up a 3 to 5 megapixel camera is fine. And if all you print is 2" to 4" images, then 3 megapixel is just fine. You only need more pixels if you plan to make poster-size enlargements. In any case, get a camera designed to fit on a camera mount on the microscope. A jury-rigged camera setup will never give you the same quality - even if the camera and microscope are good quality. Tim Morken Lab Vision / NeoMarkers www.labvision.com -----Original Message----- From: Mikael Niku [mailto:mikael.niku@helsinki.fi] Sent: Monday, October 27, 2003 11:57 PM To: grantgd@comcast.net; histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Digital Photomicroscopy Gordon, we have a BH-2 and changed from film to digital a few ears ago. We first tried a few "normal" digital cameras the Nikon D1 digital SLR, for example), but it seemed o be difficult to attach the camera to the scope properly you probably need an expert adaptor company to do his, at least with the old scopes). With the cameras specifically designed for microscope work there were no major problems, and we finally got a Colorview12 camera + image analysis software by Soft Imaging System. With a resolution of 1280x1024 pixels and less than perfect color reproduction, there are better cameras available today. But even this IS enough to produce decent publication images (the minimum resolution most journals ask is 150-200 dpi, so you can produce images at least 16 cm wide, which is of course much larger than most images in scientific publications). I actually won a honorable mention in the Nikon international photomicrography competition last year using this camera, so it can't be that bad. I never used film since getting the digital camera; the lot of slide film I bought slightly before is still sitting in the freezer. It is true that the resolution of even the best digital cameras is theoretically less than that of film. But for most scientific publishing, there is no practical difference (see above). If I understand correctly, the resolution of modern digital cameras is actually so high that at least with moderate to high magnification the limiting factor is the resolution of the microscope objective, not the camera. So you don't really get any extra information using film. My view is that with digital equipment one can produce at least as good quality images as with film, but digital is a LOT easier and quicker to use. You must note, though, that to get really optimal results, you need to spend some time learning digital color reproduction and image processing. But this is no different with film. +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ + Mikael Niku + University of Helsinki, Dept. Basic Veterinary Sciences + URL: www.helsinki.fi/~mniku/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! (Gandhi) > -----Original Message----- > From: histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of > Gordon Grant > Sent: 27. lokakuuta 2003 16:59 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] RE: Digital Photomicroscopy > > > Hello histonetters, > I have an Olympus BH-2 microscope fitted with a real nice > PM10AD Olympus film camera. I have been real happy with the > set up as I take a lot of publication photos. My problem is > digital is now the way to go. I borrowed a Coolpix MDC lens > adapter with a coolpix and Olympus C-5050 camera. I not very > pleased with the results. Does anyone have a recommendation for me. > > Best > Gordon Grant _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DRG <@t> Stowers-Institute.org Tue Oct 28 11:13:13 2003 From: DRG <@t> Stowers-Institute.org (Grant, Debra) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] embedding L's Message-ID: Thank you to all who responded to my question, I have ordered the L's from Thermo- Shandon. Thanks again, I know I can always count on you guys! Debby Grant Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, MO 64110 drg@stowers-institute.org From jqb7 <@t> cdc.gov Tue Oct 28 11:23:56 2003 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Grocott's stain for fungi Message-ID: And I do chemically clean the glassware used for the methenamine since it is a component of the methenamine-silver. Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: Jackie.O'Connor@abbott.com [mailto:Jackie.O'Connor@abbott.com] Sent: Tuesday, October 28, 2003 12:02 PM To: Kathleen Roberts Cc: Histonet Subject: Re: [Histonet] Grocott's stain for fungi Make sure you are using the correct chemical, it should be Hexamethylenetetramine (methenamine) (C6H12N4). It should dissolve readily in H20. Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics Kathleen Roberts Sent by: histonet-admin@lists.utsouthwestern.edu 10/28/2003 09:51 AM To: Histonet cc: Subject: [Histonet] Grocott's stain for fungi Hi all- I'm making up the various stock & working solutions for Grocott's stain, and I am having a heck of a time getting the 3% methenamine to go into solution. (Not methenamine silver, just plain old 3% methenamine in distilled water.) I know that you have to acid-clean the glassware for the methenamine silver & silver nitrate solutions, but do I have to do it for 3% methenamine? I tried heating it, but that did not make a bit of difference. AFIP manual has no other instructions other than "3 g methenamine in 100 ml distilled water". Is there a shelf life for methenamine? The bottle we have is pretty old, but I am not sure if that would be a factor. I intend to try acid-cleaning a bottle for the 3% methenamine this afternoon, but I wanted to see if anybody had any other suggestions to try as well. Thanks in advance- Kathleen Roberts Principal Lab Technician Neurotoxicology Labs Dept of Pharmacology & Toxicology Ernest Mario School of Pharmacy Rutgers University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/71429ef4/attachment.htm From jqb7 <@t> cdc.gov Tue Oct 28 11:26:35 2003 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] RE: PAM MARCUM Message-ID: I agree with Pam. "Junk" refers to email that you cannot be readily identified as legitimate without opening. It makes everyone's life easier if the subject line is used so your questions can be easily responded to and not deleted as unread. Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: Pamela Marcum [mailto:pmarcum@polysciences.com] Sent: Tuesday, October 28, 2003 11:45 AM To: JCarpenter764@aol.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: PAM MARCUM I think you misunderstood we get them (junk e-mails ans spam) from many sources and they are primarily from e-mails with no subject line that look official. I have no problem answering your questions or helping. The reference to garbage and junk mail was not about your questions just repeated failure to use a subject line heading. It is a simple request and after deleting a pile of e-mails this morning from you and several others I made a request that is only etiquette. As to your questions they have been very simple and should be easy to find in a library or good histology test. If you do not have these available and require help anyone on Histonet will attempt to help you. We can also suggest texts that may assist with your studies if you wish. Pam Marcum -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of JCarpenter764@aol.com Sent: Tuesday, October 28, 2003 11:27 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: PAM MARCUM OK.....i understand about the subject lines. But i have been studying and working very hard at preparing myself for the exam this summer....I do appreciate every little bit of help that i have received and don't take any of it for granted. There are some basic questions that confuse me and i thought it was ok to ask for help. I don't consider this garbage, nor do i consider this junk email. Thank you for your time -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/3dcc2d28/attachment.htm From mprice26 <@t> juno.com Tue Oct 28 11:33:24 2003 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] RE: Microwave Tissue Processors Message-ID: <20031028.093433.4667.295397@webmail21.nyc.untd.com> Hi histonetters, I am trying to rech Gwen Goss @ Cleveland Clinic in Cleveland, Ohio. A phone # or e-mail address would be great. 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 mprice26 <@t> juno.com Tue Oct 28 11:33:55 2003 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] RE: Gwen Goss Message-ID: <20031028.093524.4667.295424@webmail21.nyc.untd.com> Hi histonetters, I am trying to rech Gwen Goss @ Cleveland Clinic in Cleveland, Ohio. A phone # or e-mail address would be great. 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 tpmorken <@t> labvision.com Tue Oct 28 11:52:33 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Dionized vs distilled water Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CC6B@usca0082k03.rallansci.apogent.com> Distilled water is classically produced by heating water to evaporation and subsequent condensing on a cold surface. In the process most impurities are either evaporated off ahead of the water (in the case of most organics), or left behind (in the case of minerals). The water is also effectively deionized because the salts are left behind. It is fairly pure water. To get very pure water it needs to be re-distilled several times. Deionized water is classically passed through a salt bed or ionized resin bed that captures the mineral ions (ie, a "water softener"). The water is not necessarily pure, however, especially in regards to organic chemicals. Reverse osmosis is also used now days to deionize water. High quality water systems these days are some combination of filters, distillation, deionizing resins and reverse osmosis. Tim Morken -----Original Message----- From: JCarpenter764@aol.com [mailto:JCarpenter764@aol.com] Sent: Tuesday, October 28, 2003 7:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) while studying for my exam on the different fixatives and there ingredients....i have noticed that some call for distilled water and some use the term deionized water. Is there a difference? -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/be60c6a0/attachment.htm From juan.gutierrez <@t> christushealth.org Tue Oct 28 11:52:36 2003 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] (no subject) Message-ID: Three water molecules and a heavier molecular weight. -----Original Message----- From: JCarpenter764@aol.com [mailto:JCarpenter764@aol.com] Sent: Mon 10/27/2003 8:32 AM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] (no subject) what is the difference between the anhydrous compound and the trihydrate compound? From juan.gutierrez <@t> christushealth.org Tue Oct 28 11:53:34 2003 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] (no subject) Message-ID: Look under white blood cells, or leucocytes. -----Original Message----- From: JCarpenter764@aol.com [mailto:JCarpenter764@aol.com] Sent: Mon 10/27/2003 8:35 AM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] (no subject) can anyone explain eosinophil to me From garygill <@t> dcla.com Tue Oct 28 12:03:29 2003 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Dionized vs distilled water Message-ID: Last I read, no one has ever seen 100% pure water. Gary Gill -----Original Message----- From: Morken, Tim - Labvision [mailto:tpmorken@labvision.com] Sent: Tuesday, October 28, 2003 12:53 PM To: 'JCarpenter764@aol.com'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Dionized vs distilled water Distilled water is classically produced by heating water to evaporation and subsequent condensing on a cold surface. In the process most impurities are either evaporated off ahead of the water (in the case of most organics), or left behind (in the case of minerals). The water is also effectively deionized because the salts are left behind. It is fairly pure water. To get very pure water it needs to be re-distilled several times. Deionized water is classically passed through a salt bed or ionized resin bed that captures the mineral ions (ie, a "water softener"). The water is not necessarily pure, however, especially in regards to organic chemicals. Reverse osmosis is also used now days to deionize water. High quality water systems these days are some combination of filters, distillation, deionizing resins and reverse osmosis. Tim Morken -----Original Message----- From: JCarpenter764@aol.com [mailto:JCarpenter764@aol.com] Sent: Tuesday, October 28, 2003 7:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) while studying for my exam on the different fixatives and there ingredients....i have noticed that some call for distilled water and some use the term deionized water. Is there a difference? -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/ff4860b9/attachment.htm From Ronnie_Houston <@t> bshsi.com Tue Oct 28 12:05:45 2003 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Dionized vs distilled water Message-ID: <530361BF03351B4CAE5270A05D3037B5FE04D0@bsrexms01.BSHSIR.COM> What quality of water is recommended/regulated for anatomic and clinical pathology labs? Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 ronnie_houston@bshsi.com -----Original Message----- From: Morken, Tim - Labvision [mailto:tpmorken@labvision.com] Sent: Tuesday, October 28, 2003 12:53 PM To: 'JCarpenter764@aol.com'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Dionized vs distilled water Distilled water is classically produced by heating water to evaporation and subsequent condensing on a cold surface. In the process most impurities are either evaporated off ahead of the water (in the case of most organics), or left behind (in the case of minerals). The water is also effectively deionized because the salts are left behind. It is fairly pure water. To get very pure water it needs to be re-distilled several times. Deionized water is classically passed through a salt bed or ionized resin bed that captures the mineral ions (ie, a "water softener"). The water is not necessarily pure, however, especially in regards to organic chemicals. Reverse osmosis is also used now days to deionize water. High quality water systems these days are some combination of filters, distillation, deionizing resins and reverse osmosis. Tim Morken -----Original Message----- From: JCarpenter764@aol.com [mailto:JCarpenter764@aol.com] Sent: Tuesday, October 28, 2003 7:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) while studying for my exam on the different fixatives and there ingredients....i have noticed that some call for distilled water and some use the term deionized water. Is there a difference? ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ 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. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/06d79afb/attachment.htm From kgrobert <@t> rci.rutgers.edu Tue Oct 28 12:19:19 2003 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Grocott's stain for fungi In-Reply-To: References: Message-ID: <3F9EB327.7020502@rci.rutgers.edu> Yes, that is what I am trying to dissolve-I just wrote "methenamine" because it's an alternative name for hexamethylenetetramine and it's easier to type. :o) Plus, that's what the AFIP manual calls it. Thanks! Kathleen Rutgers University Jackie.O'Connor@abbott.com wrote: > > Make sure you are using the correct chemical, it should be > Hexamethylenetetramine (methenamine) (C6H12N4). It should dissolve > readily in H20. > > Jacqueline M. O'Connor HT(ASCP) > Abbott Laboratories > Global Pharmaceutical Research and Development > Discovery Chemotheraputics > > > > > Kathleen Roberts > Sent by: histonet-admin@lists.utsouthwestern.edu > > 10/28/2003 09:51 AM > > > To: Histonet > cc: > Subject: [Histonet] Grocott's stain for fungi > > > > > Hi all- > > I'm making up the various stock & working solutions for Grocott's stain, > and I am having a heck of a time getting the 3% methenamine to go into > solution. (Not methenamine silver, just plain old 3% methenamine in > distilled water.) I know that you have to acid-clean the glassware for > the methenamine silver & silver nitrate solutions, but do I have to do > it for 3% methenamine? I tried heating it, but that did not make a bit > of difference. AFIP manual has no other instructions other than "3 g > methenamine in 100 ml distilled water". > > Is there a shelf life for methenamine? The bottle we have is pretty > old, but I am not sure if that would be a factor. > > I intend to try acid-cleaning a bottle for the 3% methenamine this > afternoon, but I wanted to see if anybody had any other suggestions to > try as well. > > Thanks in advance- > Kathleen Roberts > Principal Lab Technician > Neurotoxicology Labs > Dept of Pharmacology & Toxicology > Ernest Mario School of Pharmacy > Rutgers University > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/3825e685/attachment.htm From ryaskovich <@t> dir.nidcr.nih.gov Tue Oct 28 12:15:30 2003 From: ryaskovich <@t> dir.nidcr.nih.gov (Yaskovich, Ruth A (NIH/NIDCR)) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] (no subject) Message-ID: <8F3AB322628548428A992EFB0E80F5D3A2EE12@nihexchange8.nih.gov> Pam, The Histonet is for anyone especially someone new to this field. I agree there are no stupid questions. I don't even have the time to check subject lines. The matter of subject lines could have been answered. There was no reason to speak down to someone. You work for a company and are lucky to be a part of Histonet, there are many Technicians on here that don't want sales reps.period. Ruth Yaskovich N.I.H. N.I.D.C.R 301-496-1392 -----Original Message----- From: Pamela Marcum [mailto:pmarcum@polysciences.com] Sent: Tuesday, October 28, 2003 11:05 AM To: JCarpenter764@aol.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] (no subject) Please use a subject line in e-mails and also where are you working from. Many of your questions are extremely basic and should have been answered in test or training. I am sorry we have had such a flurry of e-mails with no subject line that are garbage or junk e-mail list so at this point a subject line is becoming necessary if you want an answer. Pam Marcum -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of JCarpenter764@aol.com Sent: Tuesday, October 28, 2003 10:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) while studying for my exam on the different fixatives and there ingredients....i have noticed that some call for distilled water and some use the term deionized water. Is there a difference? -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/4aa763f6/attachment.htm From kgrobert <@t> rci.rutgers.edu Tue Oct 28 12:22:49 2003 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Grocott's stain for fungi In-Reply-To: References: Message-ID: <3F9EB3F9.4060203@rci.rutgers.edu> Which is what I thought as well, but still no go. :oP My boss & I have decided to order fresh hexamethylenetetramine & see if that makes any difference, as well as acid-clean the glassware. Thanks for your input! Not my day- Kathleen Rutgers University DeLovino, Salvacion S. wrote: >Preparing 3% methenamine is straightforward. No heat needed. It melts right >away in DI water. > >Salvacion S. Delovino > >-----Original Message----- >From: Kathleen Roberts [mailto:kgrobert@rci.rutgers.edu] >Sent: Tuesday, October 28, 2003 7:51 AM >To: Histonet >Subject: [Histonet] Grocott's stain for fungi > >Hi all- > >I'm making up the various stock & working solutions for Grocott's stain, >and I am having a heck of a time getting the 3% methenamine to go into >solution. (Not methenamine silver, just plain old 3% methenamine in >distilled water.) I know that you have to acid-clean the glassware for >the methenamine silver & silver nitrate solutions, but do I have to do >it for 3% methenamine? I tried heating it, but that did not make a bit >of difference. AFIP manual has no other instructions other than "3 g >methenamine in 100 ml distilled water". > > Is there a shelf life for methenamine? The bottle we have is pretty >old, but I am not sure if that would be a factor. > >I intend to try acid-cleaning a bottle for the 3% methenamine this >afternoon, but I wanted to see if anybody had any other suggestions to >try as well. > >Thanks in advance- >Kathleen Roberts >Principal Lab Technician >Neurotoxicology Labs >Dept of Pharmacology & Toxicology >Ernest Mario School of Pharmacy >Rutgers University > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From ameeker <@t> mail.jhmi.edu Tue Oct 28 12:25:03 2003 From: ameeker <@t> mail.jhmi.edu (ALAN MEEKER) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] staining mouse tissues using mouse monoclonals Message-ID: <36e703e36e44ca.36e44ca36e703e@jhmimail.jhmi.edu> Hi, I am trying immunohistochem on mouse tissues using a mouse IgG monoclonal. The target is nuclear, and I can see the signals fairly well. The problem is in background staining from the endogenous mouse antibody, especially in things like plasma cells in the lamina propria of the large intestine. I am wondering what is the best way to attenuate or eliminate this background signal. I tried something simple, namely pre-treating the tissue with an excess of goat anti-mouse IgG, hoping to bind/cover up most of the resident IgG before applying my primary and then HRP-anti mouse secondary. This seems not to have worked at all, perhaps because there were unused binding sites on the goat anti-mouse that bound my monoclonal primary (?). At any rate, the simple, quick fix didn't do the trick. I have heard that there are "mouse on mouse" kits available to deal with this problem. Has anyone had any experience using these? -Alan Meeker Alan Meeker, PhD Department of Pathology Division of Genitourinary Pathology Bunting-Blaustein Cancer Research Building Room 153 1650 Orleans Street Baltimore, MD 21231-1000 e-mail: ameeker@mail.jhmi.edu From REEVEML <@t> shands.ufl.edu Tue Oct 28 12:37:48 2003 From: REEVEML <@t> shands.ufl.edu (Mary Reeves) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] RE: Microwave Tissue Processors Message-ID: We microwave process all of our biopsy specimens and our fatty tissue (except lipoma). Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 >>> "Hallada, Teri" 10/28/03 11:42AM >>> I was wondering if anyone out there is using a microwave tissue processor for routine hospital tissues. Are there any regulations applicable to instituting one, ie FDA approval? Teri Hallada BS MT CT (ASCP) thallada@noch.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Tue Oct 28 13:16:20 2003 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] staining mouse tissues using mouse monoclonals Message-ID: I routinely use mouse primaries on Mouse tissue using either the ARK kit (K3954) from DAKO. I've tried the M.O.M. from Vector (they have a couple of 'flavors') and the MM Biotinylation Kit from Biocare (MMBK-G). All are equally superb for mouse on mouse, so take your pick. DAKO has a handy little Excel spreadsheet you can download for making the dilutions for the ARK -it comes in really handy, and you can print out the dilution sheet for data retention. Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics ALAN MEEKER Sent by: histonet-admin@lists.utsouthwestern.edu 10/28/2003 12:25 PM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] staining mouse tissues using mouse monoclonals Hi, I am trying immunohistochem on mouse tissues using a mouse IgG monoclonal. The target is nuclear, and I can see the signals fairly well. The problem is in background staining from the endogenous mouse antibody, especially in things like plasma cells in the lamina propria of the large intestine. I am wondering what is the best way to attenuate or eliminate this background signal. I tried something simple, namely pre-treating the tissue with an excess of goat anti-mouse IgG, hoping to bind/cover up most of the resident IgG before applying my primary and then HRP-anti mouse secondary. This seems not to have worked at all, perhaps because there were unused binding sites on the goat anti-mouse that bound my monoclonal primary (?). At any rate, the simple, quick fix didn't do the trick. I have heard that there are "mouse on mouse" kits available to deal with this problem. Has anyone had any experience using these? -Alan Meeker Alan Meeker, PhD Department of Pathology Division of Genitourinary Pathology Bunting-Blaustein Cancer Research Building Room 153 1650 Orleans Street Baltimore, MD 21231-1000 e-mail: ameeker@mail.jhmi.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/63033d22/attachment.htm From garygill <@t> dcla.com Tue Oct 28 13:26:46 2003 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] RE: [Microscopy] Looking for old article Message-ID: Cowen T, Haven AJ, Burnstock G. Pontamine sky blue: a counterstain for background autofluorescence in fluorescence and immunofluorescence histochemistry. Histochemistry. 1985;82(3):205-8. The stain pontamine sky blue (PSB) has been shown to reduce background autofluorescence in catecholamine fluorescence and immunofluorescence histochemical preparations. Using PSB as a counterstain on whole-mount stretch preparations of human mesenteric blood vessels, a medium dense noradrenergic nerve plexus is clearly revealed, which previously had been only partially visible because of background autofluorescence. Image analysis of nerve densities in whole-mount stretch preparations of guinea-pig arteries containing noradrenergic, substance P-, and vasoactive intestinal polypeptide (VIP)-positive nerve plexuses shows that PSB staining does not alter the specific neuronal fluorescence and that it improves image definition. If you subscribe to Loansome Doc, you can order a pdf copy for a price. Gary Gill -----Original Message----- From: Sven Terclavers [mailto:Sven.Terclavers@med.kuleuven.ac.be] Sent: Tuesday, October 28, 2003 11:55 AM To: Microscopy@msa.microscopy.com; histonet@lists.utsouthwestern.edu Subject: [Microscopy] Looking for old article ---------------------------------------------------------------------------- -- The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html ---------------------------------------------------------------------------- --- Hi all, I'm looking for a pdf or word-document of the following qrticle. Can anyone help me out? Thanks! Histochemistry. 1985;82(3):205-8. Cowen T, Haven AJ, Burnstock G. Pontamine sky blue: a counterstain for background autofluorescence in fluorescence and immunofluorescence histochemistry. Sven Terclavers From notpessimistic <@t> msn.com Tue Oct 28 14:13:01 2003 From: notpessimistic <@t> msn.com (Eric Boyd) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] subscribe digest Message-ID: -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/682dbd84/attachment.htm From ameeker <@t> mail.jhmi.edu Tue Oct 28 13:59:52 2003 From: ameeker <@t> mail.jhmi.edu (ALAN MEEKER) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] eliminating tissue autofluorescence Message-ID: <37081b8370abe4.370abe437081b8@jhmimail.jhmi.edu> Hi all. I am a newcomer to histonet and first would like to express my thanks for the many helpful responses I received to a question I posted earlier. Here's another question - we are doing a lot of immunofluorescence on formalin-fixed human tissue sections lately. We get quite a bit of background autofluorescence in several of the channels, on multiple tissue types, and we'd obviously like to avoid this. I came across a paper by a German group (sorry, don't have the reference handy at the moment) in which they claimed that by pre-bleaching the unstained slides using a medium-wattage light source (basically an aquarium grow-light)prior to staining they could eliminate much of this background fluorescence in the tissues. This seemed to make sense, so I tried several times with several different lamps but saw no real improvement, even with very long exposure times (days!). I even went so far as to remove our high-intensity light source from the fluorescence microscope and used this by exposing slides directly to the mercury lamp for times up to 30 minutes. Still no diminution of background signals. Perhaps re-hydrating the t issue first will help, as photobleaching is, I believe, supposed to be mediated by the generation of free radical species and I'll try this next. Still, I am suprised that a close-up 30 minute exposure to the unfiltered mercury arc lamp didn't do anything... Anyone else have any thoughts?? -Alan Meeker Alan Meeker, PhD Department of Pathology Division of Genitourinary Pathology Bunting-Blaustein Cancer Research Building Room 153 1650 Orleans Street Baltimore, MD 21231-1000 e-mail: ameeker@mail.jhmi.edu From lao_ji <@t> yahoo.com Tue Oct 28 16:26:48 2003 From: lao_ji <@t> yahoo.com (Jimmy Lao) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Embryo sticking in InsituPro Run Message-ID: <20031028222648.43703.qmail@web12505.mail.yahoo.com> Hi Any one can help preventing embryo sticking to the Column during InsituPro run? InsituPro is automated machine for whole mount embryo in situ hybridization. It did not happen when I used fresh column. But it did in the 2nd time using column. I guess there are something staying in the column after first use, even I treated them with H2O2 overnight and 4%PFA cleaning program treatment. How can I clean used column? Thanks! Jimmy __________________________________ Do you Yahoo!? Exclusive Video Premiere - Britney Spears http://launch.yahoo.com/promos/britneyspears/ From Kristen.Rothammer <@t> stjude.org Tue Oct 28 16:53:03 2003 From: Kristen.Rothammer <@t> stjude.org (Rothammer, Kristen) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] GFP antibodies Message-ID: <1E0CC447E59C974CA5C7160D2A2854EC459F40@SJMEMXMB04.stjude.sjcrh.local> Hello Everyone- I am desperately, and rather unsuccessfully, trying to find a better way to look at eGFP in frozen kidney sections from mice. I have read a wealth of info on histonet regarding use of GFP antibodies, and I have tried a number of methods, but I am still not having any luck. I have tried the following: (1) looking directly at eGFP in frozen sections (sections are just a few weeks old) right after 15 min fixation with 4% paraformaldehyde (fresh 16% paraformaldehyde diluted in PBS), followed by washes in PBS--> no GFP is observed (2) looking directly at eGFP in same frozen sections without fixation, coverslipping in PBS --> no GFP is observed (3) fixation in paraformaldehyde as described above followed by a GFP antibody conjugated to a fluorochrome --> very little signal is observed (4) same fixation, followed by incubation with GFP antibody, follwed by biotinylated anti-rabbit, and finally by streptavidin-Alexa 568 * This method resulted in EXTREMELY high background. I tried to block endogenous biotin by using both the egg/milk protocol and biotin/avidin drops from Vector Labs, both with little success. I mount my slides with Flouromount, do NOT seal with nail polish, and I do not do antigen retrieval (this is not necessary for frozen sections, right?). Any input would be greatly appreciated! Thank you! Kristen Rothammer St. Jude Children's Research Hospital Department of Biochemistry Kristen.rothammer@stjude.org -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/9033d042/attachment.htm From d.catmull <@t> latrobe.edu.au Tue Oct 28 16:56:04 2003 From: d.catmull <@t> latrobe.edu.au (Deanne Catmull) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] LMD ... IHC sections falling off In-Reply-To: <26588589.1066946917921.JavaMail.SYSTEM@onestop9> Message-ID: <3.0.6.32.20031029095604.008ea540@pop.latrobe.edu.au> Dear Victoria, We always use Vectabond treated slides for immunohistochemistry. Sections don't tend to float off. It's a simple procedure and good to use for pretty much any stain. Deanne at La Trobe Bundoora At 05:08 PM 10/23/03 -0500, mrl0627@mail.ecu.edu wrote: > > I sometimes lose sections (4 um thick, mounted on superfrost-plus) during the counterstaining-clearing process although they survive immuno and development in DAB. Any thoughts of what I could be doing wrong? >It's a heart breaker to lose them at the very end of the process. I am in a university lab and do everything by hand. Thanks, Maureen at ECU >-----Original Message----- >From: Amos Brooks <amosbrooks@earthlink.net> >To: histonet@lists.utsouthwestern.edu >Sent: Mon, 20 Oct 2003 16:20:39 -0400 >Subject: [Histonet] LMD ... IHC sections falling off > >Victoria, > Firstly try thinner sections 10 um is a bit thick. Next you should >use some type of charged, silanized or APES slides. You can either silanize >your own slides or buy them from a vendor. Adhesives aren't necessarily >good for IHC as they can interfere with the reaction. >Amos Brooks > > > >>Message: 5 >>From: victoria.hahn-stromberg@orebroll.se >>To: histonet@lists.utsouthwestern.edu >>Date: Mon, 20 Oct 2003 13:20:39 +0200 >>Subject: [Histonet] LMD >> >>Hi! >>I=B4m having problems with my LMD slides. I use 10um paraffin embedded >>histological sections which I would like to stain immunohistochemically = >>with >>CAM, but the sections keep falling off, can anyone plesse help = >>me??/Victoria >> >>Victoria Hahn-Str=F6mberg >>MSc, PhD student >>Kliniken f=F6r Patologi >>Universitetssjukhuset =D6rebro >>701 85 =D6rebro >>tel: 019-6023644 >>faxnr:019-6021035 > > >_______________________________________________ >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 > > Deanne Catmull Neuroimmunology Laboratory Dept of Biochemistry La Trobe University Bundoora Vic 3086 Ph:9479-1155 From DRG <@t> Stowers-Institute.org Tue Oct 28 17:03:02 2003 From: DRG <@t> Stowers-Institute.org (Grant, Debra) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Embryo sticking in InsituPro Run Message-ID: Hi Jimmy, We use 1% NaOH (Sodium Hydroxide) 6 hours to overnight, then we rinse 3x with distilled water, then soak them in distilled H20 overnight, then rinse 2x with distilled H20, then dry them in an oven @ 60 degrees Celsius, and rinse in 100% EtOH then dry again in the oven @ 60 degrees Celsius. Usually we can use the columns until the filters pop out from the columns, approximately 3-4 times. Also, if you use tween 20 at 0.1% in all hybridization solutions you shouldn't have this problem with the embryos sticking to the columns. Debby Grant Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, MO 64110 drg@stowers-institute.org -----Original Message----- From: Jimmy Lao [mailto:lao_ji@yahoo.com] Sent: Tuesday, October 28, 2003 4:27 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Embryo sticking in InsituPro Run Hi Any one can help preventing embryo sticking to the Column during InsituPro run? InsituPro is automated machine for whole mount embryo in situ hybridization. It did not happen when I used fresh column. But it did in the 2nd time using column. I guess there are something staying in the column after first use, even I treated them with H2O2 overnight and 4%PFA cleaning program treatment. How can I clean used column? Thanks! Jimmy __________________________________ Do you Yahoo!? Exclusive Video Premiere - Britney Spears http://launch.yahoo.com/promos/britneyspears/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From d.catmull <@t> latrobe.edu.au Tue Oct 28 17:08:54 2003 From: d.catmull <@t> latrobe.edu.au (Deanne Catmull) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] (no subject) In-Reply-To: <123.26c65b9b.2cce8741@aol.com> Message-ID: <3.0.6.32.20031029100854.008f1540@pop.latrobe.edu.au> Hi! Eosinophils are cells of the innate immune system and are mainly assosciated with allergies. They are found in the least percentage of all other innate cells in a healthy human. Any good immunology text book will give you more detail. Deanne at La Trobe Bundoora. At 09:35 AM 10/27/03 EST, JCarpenter764@aol.com wrote: > can anyone explain eosinophil to me Deanne Catmull Neuroimmunology Laboratory Dept of Biochemistry La Trobe University Bundoora Vic 3086 Ph:9479-1155 From wstover <@t> vet.uga.edu Tue Oct 28 17:25:53 2003 From: wstover <@t> vet.uga.edu (Wanda Stover) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Agnor Stain Message-ID: <3F9EFB01.4080707@vet.uga.edu> Are there anyone out there that doest he Agnor? If so..we are getting a precipitate after stain completion. Anyone know what it may be? Thanks Wanda University of Georgia College Veterinary Medicine From Wijaya.Martanto <@t> chbe.gatech.edu Tue Oct 28 17:46:44 2003 From: Wijaya.Martanto <@t> chbe.gatech.edu (Martanto, Wijaya) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] (no subject) Message-ID: <5D3DB84BB6F5E548AAC9FFEDC310FD091BB9A1@chicken-boo.che.gatech.edu> PLEASE UNSUBSCRIBE ME -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/a1f62e62/attachment.htm From john.mcginley <@t> colostate.edu Tue Oct 28 18:08:28 2003 From: john.mcginley <@t> colostate.edu (John McGinley) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Cryostat purchase Message-ID: <001201c39db0$c7f12360$29255281@MCGINLEY> Hi, Our research laboratory is looking at purchasing a cryostat in the near future and we need a unit that will cut sections suitable for laser capture microdisection (LCM). Most of our work revolves around rat mammary gland and past attempts at cutting this very fatty tissue on an old IEC unit proved frustrating to say the least. Our lab has been without a cryostat for a few years, so we don't know what latest and greatest units are available. We're looking for good unit that doesn't cost an arm and leg. Does anyone have any recommendations? Also, is anyone using the Instrumedics CyroJane system for LCM? Does the transfer tape interfere with RNA recovery? Any information would be greatly appreciated. Thanks, John ---------------------------- John N. McGinley, HTL(ASCP) Cancer Prevention Lab Colorado State University Email: john.mcginley@colostate.edu From mward <@t> wfubmc.edu Tue Oct 28 13:33:34 2003 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] search for antibodies Message-ID: <61135F0455D33347B5AAE209B903A304032622E4@EXCHVS2.medctr.ad.wfubmc.edu> I have a request from a Pathologist to try out the following antibodies: Dystrophin, paired helical filaments (PHF), and tubule associated unit (tau) I am looking for vendor and also anyone who has gotten them to work. We use the NexEs and also have a Dako stainer. Thanks in advance for the help. Martha Ward Wake Forest University Baptist Medical Center From mward <@t> wfubmc.edu Tue Oct 28 13:30:11 2003 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] search for antibodies Message-ID: <61135F0455D33347B5AAE209B903A304032622E3@EXCHVS2.medctr.ad.wfubmc.edu> I have a request from our Pathologist to work up the following antibodies: Dystrophin, paired helical filaments (phf), and tubule associated unit (tau) Can anyone recommend a vendor for any or all? Also if you are currently offering these could you give me some pointers? We have the NexEs as well as the Dako stainer. Thanks in advance. Martha Ward Wake Forest University Baptist Medical Center -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031028/b4576c0d/attachment.htm From gsmith <@t> confocal.com Tue Oct 28 23:11:32 2003 From: gsmith <@t> confocal.com (Glenn Smith) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] rhodamine auto-fluor. problem Message-ID: <000701c39ddb$21c3bae0$b1d8d8ce@laptop1> In a message dated 10/24/2003 3:01:17 PM US Mountain Standard Time, Bonnie.P.Whitaker@uth.tmc.edu writes: > I have a question for you guys: A researcher here in OB/GYN is doing rhodamine-labeled fluorescent work (he didn't say what antibody) on mouse fallopian tube... he thought he was having a staining problem, but has determined that his tissue is auto-fluorescing. The tissue is frozen in OCT and fixed in acetone/methanol. What can he do to quench this? < A couple more thoughts Bonnie... Rhodamine is excited with 488nm (blue light). I am assuming the system used in this case employs such a light source. Most organic material fluoresce under 488nm excitation. The best option is to use a narrow bandpass filter (20-40nm bandwidth centered about the peak emission wavelength of the dye). This will not completely eliminate autofluorescence but it will greatly reduce it. Another possibility is to image the tissue before and after staining to see the difference. Then could subtract one image from the other... Glenn Smith, P.Eng. 519.886.9013 x38 gsmith@confocal.com Biomedical Photometrics Inc. A12-550 Parkside Dr. Waterloo, ON N2L 5V4 Widefield High Resolution Imaging Instruments & Software Please visit www.confocal.com for more information. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031029/e208d262/attachment.htm From louise_renton <@t> hotmail.com Wed Oct 29 03:41:01 2003 From: louise_renton <@t> hotmail.com (louise renton) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Re: speaking down Message-ID: Suggesting that an individual goes to a text book to answer questions is in the interest of the individual. not necesarily "talking down" While the histonet and other electronic sources provide a quick fix answer (albeit accurate), it does not IMHO, offer the same validation as referring to standard texts, which JCarpenter ought to have if he is studying for an exam ----Original Message Follows---- From: "Yaskovich, Ruth A (NIH/NIDCR)" To: 'Pamela Marcum' , JCarpenter764@aol.com, histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] (no subject) Date: Tue, 28 Oct 2003 13:15:30 -0500 Pam, The Histonet is for anyone especially someone new to this field. I agree there are no stupid questions. I don't even have the time to check subject lines. The matter of subject lines could have been answered. There was no reason to speak down to someone. You work for a company and are lucky to be a part of Histonet, there are many Technicians on here that don't want sales reps.period. Ruth Yaskovich N.I.H. N.I.D.C.R 301-496-1392 -----Original Message----- From: Pamela Marcum [mailto:pmarcum@polysciences.com] Sent: Tuesday, October 28, 2003 11:05 AM To: JCarpenter764@aol.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] (no subject) Please use a subject line in e-mails and also where are you working from. Many of your questions are extremely basic and should have been answered in test or training. I am sorry we have had such a flurry of e-mails with no subject line that are garbage or junk e-mail list so at this point a subject line is becoming necessary if you want an answer. Pam Marcum -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of JCarpenter764@aol.com Sent: Tuesday, October 28, 2003 10:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) while studying for my exam on the different fixatives and there ingredients....i have noticed that some call for distilled water and some use the term deionized water. Is there a difference? _________________________________________________________________ Chat to anyone, anywhere with MSN Messenger! http://messenger.msn.co.za/ From ian.montgomery <@t> bio.gla.ac.uk Wed Oct 29 04:16:32 2003 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:22:06 2005 Subject: Fwd: [Histonet] Cryostat purchase Message-ID: <6.0.0.22.2.20031029101057.02cf5060@udcf.gla.ac.uk> John, Contact Alan Bright, ABright@brightinstruments.co.uk He'll supply you with all the information you need. Ian. >From: "John McGinley" >To: >X-Mailer: Microsoft Outlook, Build 10.0.4510 >Importance: Normal >X-Scan-Signature: 690e9200c080ca55fd7a8d0fb0193eeb >Sender: histonet-admin@lists.utsouthwestern.edu >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.0.13 >List-Help: >List-Post: >List-Subscribe: , > >List-Id: For the exchange of information pertaining to histotechnology and >related fields >List-Unsubscribe: , > > >List-Archive: >Date: Tue, 28 Oct 2003 17:08:28 -0700 >X-Scan-Signature: 18b8f45f5f122038853a7119e027f2bd >X-SA-Exim-Mail-From: histonet-admin@lists.utsouthwestern.edu >Subject: [Histonet] Cryostat purchase >X-Spam-Checker-Version: SpamAssassin 2.60 (1.212-2003-09-23-exp) on > swlx167.swmed.edu >X-Spam-Status: No, hits=0.0 required=6.5 tests=none autolearn=no version=2.60 >X-Spam-Level: >X-SA-Exim-Version: 3.1 (built Tue Sep 9 12:31:04 CDT 2003) >X-SA-Exim-Scanned: Yes > >Hi, > >Our research laboratory is looking at purchasing a cryostat in the near >future and we need a unit that will cut sections suitable for laser capture >microdisection (LCM). Most of our work revolves around rat mammary gland and >past attempts at cutting this very fatty tissue on an old IEC unit proved >frustrating to say the least. > >Our lab has been without a cryostat for a few years, so we don't know what >latest and greatest units are available. We're looking for good unit that >doesn't cost an arm and leg. Does anyone have any recommendations? > >Also, is anyone using the Instrumedics CyroJane system for LCM? Does the >transfer tape interfere with RNA recovery? Any information would be greatly >appreciated. > >Thanks, > >John > >---------------------------- >John N. McGinley, HTL(ASCP) >Cancer Prevention Lab >Colorado State University >Email: john.mcginley@colostate.edu > > > > > >_______________________________________________ >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 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031029/6de4e64e/attachment.htm From ian.montgomery <@t> bio.gla.ac.uk Wed Oct 29 04:46:25 2003 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Sirius Red Technique. Message-ID: <6.0.0.22.2.20031029104203.02ccf1d0@udcf.gla.ac.uk> Quick question and the man himself may be able to answer. Sirius Red Technique (Llewellyn, 1970), what is the shelf life of the prepared stain? 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 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031029/b5b159da/attachment.htm From ASelf <@t> gmhsc.com Wed Oct 29 05:23:36 2003 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Prefer formalin-free fixative Message-ID: Dear Histonetters, Has anyone tried the Prefer formalin-free fixative distributed by Anatech LTD.? If so, what were you likes and dislikes about it. Thanks in advance for your responses. Amy Self Georgetown Hospital Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or 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 burch007 <@t> mc.duke.edu Wed Oct 29 07:06:17 2003 From: burch007 <@t> mc.duke.edu (James L Burchette) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Special Stain for Keratin, not IHC Message-ID: Hello All: I was asked if there was a special stain for keratin. Seems that I recall seeing it once, but can't remember where. This is not an immuno stain. A reference or procedure would be appreciated. Thanks, Jim Burchette -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031029/1dda91c7/attachment.htm From Marjorie.Lehman <@t> unilever.com Wed Oct 29 06:41:27 2003 From: Marjorie.Lehman <@t> unilever.com (marjorie lehman) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Special Stain for Keratin, not IHC Message-ID: Hi Jim, In the third edition of the AFIP manual there are 2 stains for keratin: Ayoub-Shklar method for keratin and prekeratin and Dane's method for prekeratin, keratin and mucin. I have used the A-S one and I have also used a modified Papanicaou. Both are nice -take a bit of tweaking though. Marge -----Original Message----- From: James L Burchette [SMTP:burch007@mc.duke.edu] Sent: Wednesday, October 29, 2003 8:06 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Special Stain for Keratin, not IHC Hello All: I was asked if there was a special stain for keratin. Seems that I recall seeing it once, but can't remember where. This is not an immuno stain. A reference or procedure would be appreciated. Thanks, Jim Burchette From Loralee_Gehan <@t> URMC.Rochester.edu Wed Oct 29 06:41:50 2003 From: Loralee_Gehan <@t> URMC.Rochester.edu (Gehan, Loralee) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Prefer formalin-free fixative Message-ID: <95774A6A6036D411AFEA00D0B73C864308880466@exmc3.urmc.rochester.edu> Hi Amy, We just ordered some of this stuff and we haven't worked out all of the conditions to make sure that the fixation is complete. BUT, we have fixed mouse hind limbs and tail. Some for 6 hours and some for overnight. I think that due to the size of the tissue that the over night is better. The best part of it all, it doesn't smell like formalin. I haven't had a chance to try any IHC on these samples but the company claims that you need to do little if any antigen retrieval. Our special staining has been beautiful. Nuclear detail is great. Hope that helps. Loralee Gehan Orthopaedics Research Lab University of Rochester > ---------- > From: Amy Self > Sent: Wednesday, October 29, 2003 6:23 AM > To: 'histonet@lists.utsouthwestern.edu' > Subject: [Histonet] Prefer formalin-free fixative > > > > Dear Histonetters, > > > Has anyone tried the Prefer formalin-free fixative distributed by > Anatech LTD.? If so, what were you likes and dislikes about it. > Thanks in advance for your responses. > > > Amy Self > Georgetown Hospital > > > > Note: The information contained in this message may be privileged and > confidential and protected from disclosure. If the reader of this message > is > not the intended recipient, or an employee or 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 FreidaC <@t> aol.com Wed Oct 29 07:11:52 2003 From: FreidaC <@t> aol.com (FreidaC@aol.com) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Address Message-ID: <159.26b3e533.2cd11698@aol.com> Robert Lott, please send me your email address. I do not have my regular computer hooked up yet and I wiped out a lot of addresses on my lap-top. I will then send you the instructions for the elastic critique. Freida From hadi83 <@t> comcast.net Wed Oct 29 07:47:40 2003 From: hadi83 <@t> comcast.net (Hadi Yaziji) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Prefer formalin-free fixative In-Reply-To: Message-ID: <7676D219-0A16-11D8-84F5-00039378E76A@comcast.net> Dear Amy, I don't recommend it for the following reasons: 1. It has EXTREMELY harmful effects on many antibodies and on the DNA, so if the Prefer-fixed specimen will require any IHC or FISH, the results will be severely compromised because of the fixative. 2. Contrary to what was marketed by the company that sells it, it is not proven to be safe, and not requiring the strict regulations (in terms of proper ventilation and disposal). It simply hasn't made it to the radar screen of OSHA. I work in a reference lab, and we receive specimens that are fixed in a variety of fixatives. Prefer fixative is one of the worst in terms of destroying the IHC and FISH signal. Best wishes, Hadi Yaziji, M.D. PhenoPath Laboratories On Wednesday, October 29, 2003, at 03:23 AM, Amy Self wrote: > > > Dear Histonetters, > > > Has anyone tried the Prefer formalin-free fixative distributed by > Anatech LTD.? If so, what were you likes and dislikes about it. > Thanks in advance for your responses. > > > Amy Self > Georgetown Hospital > > > > Note: The information contained in this message may be privileged and > confidential and protected from disclosure. If the reader of this > message is > not the intended recipient, or an employee or 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 pam <@t> ategra.com Wed Oct 29 08:17:42 2003 From: pam <@t> ategra.com (Pam Barker (extension 234)) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Histology openings Latest Update 10/29/03 Message-ID: Hi histonetters, I am presently on a search for 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. These 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 supervisory, team lead and bench positions. These positions require HTL or HT certification or registry eligibility. Here are my NEWEST openings: 1. Las Vegas, NV - Histo Tech 2. Minneapolis, MN - Histo Tech 3. Washington DC - Histo Tech 4. Dayton, OH - Histo Tech 5. Los Angeles - CA Histo Tech Immunohistochemistry (2 positions) 6. Hartford, CT - Histo Tech 7. Orlando, FL - Histo Tech 8. Lancaster, Pa - Histology Supervisor 9. Miami, FL - Histo Tech Here are some of my hottest Histology Supevisory positions: 1. Maine - Histology Supervisor 2. Syracuse, NY - Histology Supervisor 3. Northern NJ - Histology Supervisor or Team Lead 4. Atlanta, GA - Team Lead grossing required 5. New Hampshire - Histo Supervisor 6. Northern NJ - Histo Supervisor Here are some of my hottest Histo Tech bench positions: 1. Southwest FL - opportunity to obtain QIHC some grossing required 2. Pittsburgh, PA - Histo Tech 3. Richmond, Va - Lead Histo Tech 4. Salt Lake City, UT - Histo Tech 5. Illinois - Team Lead 6. Boulder, CO - Histo Tech 7. Oregon - Histo Tech 8. Upstate NY - Histo Tech 9. NYC, NY - Histo Tech 10. New Orleans, LA - MOHS/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 do not have an email address, if you could pass my query & name on to them I'd be very grateful. I don't want to be a bother - I was told that you were a hands-on Histo Tech or a Lab Supervisor. If you are no longer working in a lab please send me an email and I will remove you from my list of people to contact. 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: Ategra Systems Inc Specialists in Permanent & Contract Staffing VOICE: 407-671-5800 ext 234 TOLLFREE: 800-466-9919 ext 234 EMAIL: pam@ategra.com WEBSITE: --------------------------------------------------------- From pam <@t> ategra.com Wed Oct 29 08:20:03 2003 From: pam <@t> ategra.com (Pam Barker (extension 234)) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Histology openings Latest Update 10/29/03 Message-ID: Hi histonetters, I am presently on a search for 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. These 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 supervisory, team lead and bench positions. These positions require HTL or HT certification or registry eligibility. Here are my NEWEST openings: 1. Las Vegas, NV - Histo Tech 2. Minneapolis, MN - Histo Tech 3. Dayton, OH - Histo Tech 4. Los Angeles - CA Histo Tech Immunohistochemistry (2 positions) 5. Hartford, CT - Histo Tech 6. Orlando, FL - Histo Tech 7. Lancaster, Pa - Histology Supervisor 8. Miami, FL - Histo Tech Here are some of my hottest Histology Supevisory positions: 1. Maine - Histology Supervisor 2. Syracuse, NY - Histology Supervisor 3. Northern NJ - Histology Supervisor or Team Lead 4. Atlanta, GA - Team Lead grossing required 5. New Hampshire - Histo Supervisor 6. Northern NJ - Histo Supervisor Here are some of my hottest Histo Tech bench positions: 1. Southwest FL - opportunity to obtain QIHC some grossing required 2. Pittsburgh, PA - Histo Tech 3. Richmond, Va - Lead Histo Tech 4. Salt Lake City, UT - Histo Tech 5. Illinois - Team Lead 6. Boulder, CO - Histo Tech 7. Oregon - Histo Tech 8. Upstate NY - Histo Tech 9. NYC, NY - Histo Tech 10. New Orleans, LA - MOHS/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. Thank You !! Pam - 800 466 9919 ext 234 --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing Learn More About Ategra: Ategra Systems Inc Specialists in Permanent & Contract Staffing VOICE: 407-671-5800 ext 234 TOLLFREE: 800-466-9919 ext 234 EMAIL: pam@ategra.com WEBSITE: --------------------------------------------------------- From mbryhan <@t> NORTHERNHEALTH.ORG Wed Oct 29 08:18:38 2003 From: mbryhan <@t> NORTHERNHEALTH.ORG (Mary Bryhan) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Prefer formalin-free fixative Message-ID: All, I would like to go on record to dispute the claims of Hadi regarding Prefer fixative. It is much safer than formalin based fixatives for the health of the workers as well as the environment. The main reason people have trouble with working up their antibodies using Prefer fixed tissues are, in most cases, they haven't tried to work through the protocols to determine the corrective action to get their signal to light up! We have had about 8 years or so, experience with using Prefer fixative and have EXCELLENT results! If anyone out there needs to network with other Prefer users, feel free to contact myself or my coworker Ellen Clausen-Simon at (231)487-4169. Mary Bryhan HT (ASCP) Team Leader Anatomic Pathology Department Northern Michigan Hospital Petoskey, MI 49770 >>> Hadi Yaziji 10/29/03 08:47AM >>> Dear Amy, I don't recommend it for the following reasons: 1. It has EXTREMELY harmful effects on many antibodies and on the DNA, so if the Prefer-fixed specimen will require any IHC or FISH, the results will be severely compromised because of the fixative. 2. Contrary to what was marketed by the company that sells it, it is not proven to be safe, and not requiring the strict regulations (in terms of proper ventilation and disposal). It simply hasn't made it to the radar screen of OSHA. I work in a reference lab, and we receive specimens that are fixed in a variety of fixatives. Prefer fixative is one of the worst in terms of destroying the IHC and FISH signal. Best wishes, Hadi Yaziji, M.D. PhenoPath Laboratories On Wednesday, October 29, 2003, at 03:23 AM, Amy Self wrote: > > > Dear Histonetters, > > > Has anyone tried the Prefer formalin-free fixative distributed by > Anatech LTD.? If so, what were you likes and dislikes about it. > Thanks in advance for your responses. > > > Amy Self > Georgetown Hospital > > > > Note: The information contained in this message may be privileged and > confidential and protected from disclosure. If the reader of this > message is > not the intended recipient, or an employee or 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 > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cfavara <@t> niaid.nih.gov Wed Oct 29 08:30:31 2003 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Cryostat purchase Message-ID: John, I would demo every cryostat that I could as I find them somewhat user specific. I have not looked into things in a few years but do remember a cryostat I think Bright that has a separate temperature control for the specimen stage that is more finely tuned than most cryostats. I cutting fatty tissue I think that would be very desirable. I have done some guinea pig and mink skin in the past and think I would have loved to have that capacity. Good Luck! Hope you get what you need and want! c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 -----Original Message----- From: John McGinley [mailto:john.mcginley@colostate.edu] Sent: Tuesday, October 28, 2003 5:08 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cryostat purchase Hi, Our research laboratory is looking at purchasing a cryostat in the near future and we need a unit that will cut sections suitable for laser capture microdisection (LCM). Most of our work revolves around rat mammary gland and past attempts at cutting this very fatty tissue on an old IEC unit proved frustrating to say the least. Our lab has been without a cryostat for a few years, so we don't know what latest and greatest units are available. We're looking for good unit that doesn't cost an arm and leg. Does anyone have any recommendations? Also, is anyone using the Instrumedics CyroJane system for LCM? Does the transfer tape interfere with RNA recovery? Any information would be greatly appreciated. Thanks, John ---------------------------- John N. McGinley, HTL(ASCP) Cancer Prevention Lab Colorado State University Email: john.mcginley@colostate.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Ngilman <@t> nbhd.org Wed Oct 29 08:48:03 2003 From: Ngilman <@t> nbhd.org (Noreen Gilman) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Transcription/Dictation equipment Message-ID: Good morning everyone. We are looking to update our ancient dictation equipment. The ones we have now are no longer being made and are on their last legs. We are using the regular sized cassette tapes, but would like to know what other people are using. Thanks for your input. Noreen Noreen Gilman, BS, HT (ASCP) QIHC Histopathology Supervisor Broward General Medical Center Ft. Lauderdale, FL 33316 954.355.4358 Phone 954.355.4139 Fax 954.387.0213 Pager *************************************************************************************************************************************** CONFIDENTIALITY NOTICE: This message and any included attachments are from the North Broward Hospital District (NBHD) and are intended for the sole use of the individual or entity to which it is addressed. This message may contain information that is confidential and protected by federal and state law. If you are not the intended recipient, you are hereby notified that any disclosure, copying, or distribution of this message is strictly prohibited. If you received this message in error, please immediately notify the sender by reply e-mail and then delete the original message and its attachments without reading or saving the attachments in any manner. Thank you. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031029/2feb5299/attachment.htm From pdelvent <@t> wyoming.com Wed Oct 29 08:51:33 2003 From: pdelvent <@t> wyoming.com (Priscilla Delventhal) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] basic questions about purified water Message-ID: Not being a chemist, I hesitate to reply to this persons query, however, I don't feel any of the current posts really try to answer the question about distilled vs deionized water. I don't really feel that it is a basic question, as I have asked that question over the years to other technologists and have had many explainations without finding anyone who really could explain or seemed to understand. Having said that, I offer the following and hope to have my misunderstandings cleared up. It is my understanding that distilled water removes all impurities, including the salts of metals that might interfere with the metals you are trying to stain or might combine with the metals in the stains you are using. Deionization of water helps to remove all positive and negative ions from the different components in the water. I'm not sure that it is possible to remove all ions as the Hydrogen and Oxygen in the water need ions to stay combined (I can hear some of you chemists now - probably laughing at this). The basic rules of ionization are that like ions repel each other and unlike ions attract each other. A good example of this useage is in our positively charged slides that are so popular now. Having said all of that, depending upon what chemical you are using and what you are staining for, water that has all of these elements (distilled and deionized) removed would be better to use as it would prevent the unwanted combination of elements in your staining and produce a cleaner slide with the focus of your stain very clear and without precipitate. I think that distilled and deionized water are used interchangably by many people without thinking that they are two separate procedures, many times combined in water purification systems. OK Histonetters - have at it. I know that some of this is probably incorrect. I am now semi-retired and have broad shoulders. Hopefully, I have helped the person who is studying for his exam. Priscilla in Central Wyoming From msandova <@t> nd.edu Wed Oct 29 09:03:33 2003 From: msandova <@t> nd.edu (msandova@nd.edu) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Special Stain for Keratin, not IHC In-Reply-To: References: Message-ID: <1067439813.3f9fd6c5556d0@webmail.nd.edu> Ayoub-Shklar's "Keratin and prekeratin stain" - Lee Luna's "Manual of Histologic Staining Methods of the AFIP" - 3rd edition - pp. 82 Mayra Quoting James L Burchette : > Hello All: > > I was asked if there was a special stain for keratin. Seems that I recall > seeing it once, but can't remember where. > This is not an immuno stain. A reference or procedure would be > appreciated. > > Thanks, > > Jim Burchette From mrl0627 <@t> mail.ecu.edu Wed Oct 29 09:03:59 2003 From: mrl0627 <@t> mail.ecu.edu (mrl0627@mail.ecu.edu) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] When to use tap water? Message-ID: Hello, all: The prof for my graduate course in histology insisted that tap water be used for rinsing slides during certain staining procedures (eg. H&E)although he did not give a specific reason why. When no "flavor" of water is specified in a procedure, I generally use MilliQ or distilled deionized water. Is this the safest route to take or should one use tap if nothing but "water" is listed? Thanks for the attention. Maureen, MS candidate at ECU. -----Original Message----- From: "Houston, Ronnie" <Ronnie_Houston@bshsi.com> To: "'Morken, Tim - Labvision'" <tpmorken@labvision.com>, histonet@lists.utsouthwestern.edu Sent: Tue, 28 Oct 2003 13:05:45 -0500 Subject: RE: [Histonet] Dionized vs distilled water What quality of water is recommended/regulated for anatomic and clinical pathology labs? Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 ronnie_houston@bshsi.com -----Original Message----- From: Morken, Tim - Labvision [mailto:tpmorken@labvision.com] Sent: Tuesday, October 28, 2003 12:53 PM To: 'JCarpenter764@aol.com'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Dionized vs distilled water Distilled water is classically produced by heating water to evaporation and subsequent condensing on a cold surface. In the process most impurities are either evaporated off ahead of the water (in the case of most organics), or left behind (in the case of minerals). The water is also effectively deionized because the salts are left behind. It is fairly pure water. To get very pure water it needs to be re-distilled several times. Deionized water is classically passed through a salt bed or ionized resin bed that captures the mineral ions (ie, a "water softener"). The water is not necessarily pure, however, especially in regards to organic chemicals. Reverse osmosis is also used now days to deionize water. High quality water systems these days are some combination of filters, distillation, deionizing resins and reverse osmosis. Tim Morken -----Original Message----- From: JCarpenter764@aol.com [mailto:JCarpenter764@aol.com] Sent: Tuesday, October 28, 2003 7:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) while studying for my exam on the different fixatives and there ingredients....i have noticed that some call for distilled water and some use the term deionized water. Is there a difference? ________________________________________________________________________ ________________________________________________________ ________________________________________________________________________ ________________________________________________________ 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 jluis.palazon <@t> icman.csic.es Wed Oct 29 09:04:44 2003 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] line subject Message-ID: <20031029150444.1FEA76C3F9@perceval.uca.es> Dear histo-fellows Greetings. I am in complete agreement that all of us must try not to forget to put the subject line when sending messages to the list. Personally I delete all messages without subject. On the other hand I consider that the histonet is a specialized and excelent forum not only to help fellow histoworkers to resolve dificult problems but to help people who is begining working in this area and of course students and people who is "in training". I mean a very good educational and training tool. Personaly I have learned very much reading the questions and answers. In this respect I want to say that any question, even the easiest ones are important, because the list is integrated by people with much experience and other that are begining or trying to learn histology. Have a very good day José Luis El dia 28/10/2003 17:44 usted envio el siguiente mensaje: >Date: 28 de Octubre de 2003 17:44:59 >From: "Pamela Marcum" >Subject: RE: [Histonet] RE: PAM MARCUM >To: JCarpenter764@aol.com, histonet@lists.utsouthwestern.edu > > I think you > misunderstood we get them (junk e-mails ans spam) from many sources and they are > primarily from e-mails with no subject line that look official.  I have no > problem answering your questions or helping.  The reference to garbage and > junk mail was not about your questions just repeated failure to use a subject > line heading.  It is a simple request and after deleting a pile of e-mails > this morning from you and several others I made a request that is only > etiquette.  As to your questions they have been very simple and should be > easy to find in a library or good histology test.  If you do not have these > available and require help anyone on Histonet will attempt to help you.  We > can also suggest texts that may assist with your studies if you > wish.   Pam Marcum > -----Original Message----- From: > histonet-admin@lists.utsouthwestern.edu > [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of > JCarpenter764@aol.com Sent: Tuesday, October 28, 2003 11:27 > AM To: histonet@lists.utsouthwestern.edu Subject: > [Histonet] RE: PAM MARCUM OK.....i understand about the > subject lines.  But i have been studying and working very hard at > preparing myself for the exam this summer....I do appreciate every little bit > of help that i have received and don't take any of it for granted.  There > are some basic questions that confuse me and i thought it was ok to ask for > help. I don't consider this garbage, nor do i consider this junk > email.    Thank you for your time From tieman <@t> smtp.albany.edu Wed Oct 29 04:19:27 2003 From: tieman <@t> smtp.albany.edu (Suzannah B. Tieman) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] re: purified water Message-ID: <200310291519.h9TFJRg3010012@smtp.albany.edu> Hi all In my experience, distilled water is more likely to be free of bacteria. Su Tieman Suzannah B. Tieman tieman@albany.edu Biological Sciences, SUNY-Albany 1400 Washington Ave Albany, NY 12222 ph: (518) 442-4317 fax: (518) 442-4767 From garygill <@t> dcla.com Wed Oct 29 09:26:10 2003 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] When to use tap water? Message-ID: The pH of tap water is often higher than that for non-tap water, hence bluing of hematoxylin occurs slightly more quickly. Using "designer" water is overkill for staining applications, IMHO and experience. Gary Gill -----Original Message----- From: mrl0627@mail.ecu.edu [mailto:mrl0627@mail.ecu.edu] Sent: Wednesday, October 29, 2003 10:04 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] When to use tap water? Hello, all: The prof for my graduate course in histology insisted that tap water be used for rinsing slides during certain staining procedures (eg. H&E)although he did not give a specific reason why. When no "flavor" of water is specified in a procedure, I generally use MilliQ or distilled deionized water. Is this the safest route to take or should one use tap if nothing but "water" is listed? Thanks for the attention. Maureen, MS candidate at ECU. -----Original Message----- From: "Houston, Ronnie" <Ronnie_Houston@bshsi.com> To: "'Morken, Tim - Labvision'" <tpmorken@labvision.com>, histonet@lists.utsouthwestern.edu Sent: Tue, 28 Oct 2003 13:05:45 -0500 Subject: RE: [Histonet] Dionized vs distilled water What quality of water is recommended/regulated for anatomic and clinical pathology labs? Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 ronnie_houston@bshsi.com -----Original Message----- From: Morken, Tim - Labvision [mailto:tpmorken@labvision.com] Sent: Tuesday, October 28, 2003 12:53 PM To: 'JCarpenter764@aol.com'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Dionized vs distilled water Distilled water is classically produced by heating water to evaporation and subsequent condensing on a cold surface. In the process most impurities are either evaporated off ahead of the water (in the case of most organics), or left behind (in the case of minerals). The water is also effectively deionized because the salts are left behind. It is fairly pure water. To get very pure water it needs to be re-distilled several times. Deionized water is classically passed through a salt bed or ionized resin bed that captures the mineral ions (ie, a "water softener"). The water is not necessarily pure, however, especially in regards to organic chemicals. Reverse osmosis is also used now days to deionize water. High quality water systems these days are some combination of filters, distillation, deionizing resins and reverse osmosis. Tim Morken -----Original Message----- From: JCarpenter764@aol.com [mailto:JCarpenter764@aol.com] Sent: Tuesday, October 28, 2003 7:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) while studying for my exam on the different fixatives and there ingredients....i have noticed that some call for distilled water and some use the term deionized water. Is there a difference? ________________________________________________________________________ ________________________________________________________ ________________________________________________________________________ ________________________________________________________ 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From REEVEML <@t> shands.ufl.edu Wed Oct 29 09:37:52 2003 From: REEVEML <@t> shands.ufl.edu (Mary Reeves) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] RE: Microwave Tissue Processors Message-ID: Amy, Do you currently have a microwave tissue processor? We use the Hacker/Milestone RHS-1 Microwave to process our fatty tissue. The times that you see only represent the time at temperature, they do not include the "ramp" up time (the time needed to bring the solution to the appropriate processing temperature). Fixation - We fix the tissue cassettes overnight (6pm - 3am) in Alcoholic Zinc Formalin Ethanol - 10 minutes at 65 C JFC - Produced by Hacker. Isopropanol can be used instead, however, you may need to extend the time at temperature. - 1 hour and 37 minutes at 68 C Vaporization (Vacuum Drying) -Pressure at 500 mbar Paraffin - 48 minutes at 65 C and pressure at 100 mbar This program takes approximately 4 hours to run, it replaces our 16 hour overnight program that we previously used to process fatty tissue. The only fatty tissue we do not process in the microwave is lipoma. Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 >>> Amy Self 10/28/03 01:58PM >>> Mary, Could you share with me your procedure for processing fatty tissue in the microwave. Thanks, Amy Self -----Original Message----- From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] Sent: Tuesday, October 28, 2003 1:38 PM To: histonet@lists.utsouthwestern.edu; thallada@noch.org Subject: RE: [Histonet] RE: Microwave Tissue Processors We microwave process all of our biopsy specimens and our fatty tissue (except lipoma). Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 >>> "Hallada, Teri" 10/28/03 11:42AM >>> I was wondering if anyone out there is using a microwave tissue processor for routine hospital tissues. Are there any regulations applicable to instituting one, ie FDA approval? Teri Hallada BS MT CT (ASCP) thallada@noch.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or 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 hadi83 <@t> comcast.net Wed Oct 29 09:45:33 2003 From: hadi83 <@t> comcast.net (Hadi Yaziji) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Prefer formalin-free fixative In-Reply-To: Message-ID: I measure my words! You said you had "EXCELLENT results!" with Prefer. If you have been using FISH on Prefer-fixed specimens, so please enlighten the CAP, Vysis, myself and others (to mention another pathologist, Dr. Ray Tubbs at the Cleveland Clinic) as to how you got FISH to work on these specimen. Prefer is listed by Vysis/Abbott as one of the fixatives incompatible with FISH. Also, Prefer DOES significantly reduce the signal of MANY essential antibodies including ER, Ki-67 to mention a few. You also said "It is much safer than formalin". In order to judge that something is safe, please provide clinical follow-up studies of the LONG-TERM effects of Prefer on a sizable population of histology lab workers. I look forward to your reply. Hadi Yaziji, M.D. PhenoPath Laboratories On Wednesday, October 29, 2003, at 06:18 AM, Mary Bryhan wrote: > All, > > I would like to go on record to dispute the claims of Hadi regarding > Prefer fixative. It is much safer than formalin based fixatives for > the health of the workers as well as the environment. The main reason > people have trouble with working up their antibodies using Prefer > fixed tissues are, in most cases, they haven't tried to work through > the protocols to determine the corrective action to get their signal > to light up! We have had about 8 years or so, experience with using > Prefer fixative and have EXCELLENT results! If anyone out there needs > to network with other Prefer users, feel free to contact myself or my > coworker Ellen Clausen-Simon at (231)487-4169. > > Mary Bryhan HT (ASCP) > Team Leader Anatomic Pathology Department > Northern Michigan Hospital > Petoskey, MI 49770 > >>>> Hadi Yaziji 10/29/03 08:47AM >>> > Dear Amy, > > I don't recommend it for the following reasons: > 1. It has EXTREMELY harmful effects on many antibodies and on the DNA, > so if the Prefer-fixed specimen will require any IHC or FISH, the > results will be severely compromised because of the fixative. > 2. Contrary to what was marketed by the company that sells it, it is > not proven to be safe, and not requiring the strict regulations (in > terms of proper ventilation and disposal). It simply hasn't made it to > the radar screen of OSHA. > > I work in a reference lab, and we receive specimens that are fixed in a > variety of fixatives. Prefer fixative is one of the worst in terms of > destroying the IHC and FISH signal. > > Best wishes, > > Hadi Yaziji, M.D. > PhenoPath Laboratories > > On Wednesday, October 29, 2003, at 03:23 AM, Amy Self wrote: > >> >> >> Dear Histonetters, >> >> >> Has anyone tried the Prefer formalin-free fixative distributed by >> Anatech LTD.? If so, what were you likes and dislikes about it. >> Thanks in advance for your responses. >> >> >> Amy Self >> Georgetown Hospital >> >> >> >> Note: The information contained in this message may be privileged and >> confidential and protected from disclosure. If the reader of this >> message is >> not the intended recipient, or an employee or 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 >> > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From REEVEML <@t> shands.ufl.edu Wed Oct 29 09:48:09 2003 From: REEVEML <@t> shands.ufl.edu (Mary Reeves) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] RE: Microwave Tissue Processors Message-ID: We use the RHS-1 (Hacker/Milestone) with vacuum to process fatty tissue and the RHS-2 (Hacker/Milestone) to process biopsies, . We ran duplicate blocks (one for the microwave and one for the VIP processor) for the majority of tissue types. The medical director was given the H&Es for a side by side comparison. We also tested our most common IHC stains and DNA ploidy on the microwave processed tissue. After the medical director reviewed all of the slides we were given the green light. Many of the pathologist did not even know about the switch and could not see a difference. Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 >>> "Hallada, Teri" 10/28/03 02:01PM >>> Mary, What instrument are using? Did you have to perform correlation studies? And finally, how did the pathologists do with the change? teri > -----Original Message----- > From: Mary Reeves [SMTP:REEVEML@shands.ufl.edu] > Sent: Tuesday, October 28, 2003 13:38 > To: histonet@lists.utsouthwestern.edu; Hallada, Teri > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > We microwave process all of our biopsy specimens and our fatty tissue (except lipoma). > > Mary Reeves > Technical Specialist > Histology > 1-352-265-0680 ext 7-2113 > > >>> "Hallada, Teri" 10/28/03 11:42AM >>> > I was wondering if anyone out there is using a microwave tissue processor for routine hospital tissues. Are there any regulations applicable to instituting one, ie FDA approval? > Teri Hallada BS MT CT (ASCP) > thallada@noch.org > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From DonnaWillis <@t> texashealth.org Wed Oct 29 10:00:27 2003 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] RE: Microwave Tissue Processors Message-ID: <5C6CBCCEB04B894C8BD15B312487F7B2205A7A@ftwex01.txhealth.org> Teri, Here at Harris we have programs for Biopsy size samples and a program for 5mm thick samples that are used daily. We also still use routine tissue processors. The main thing to consider is that you should be using a laboratory microwave not a household unit. Household units are not made for laboratory use. There are several laboratory vendors available and procedures must be optimized taking into consideration instrumentation, specimen, container, reagents and even geography (altitude). I will decrease turn-around-time and reagent cost. Contact the vendors and ask for a demonstration. Milestone, Energy Beam, Sakura, Richard Allan, EMS, Pella are a few to consider. If there are other vendors that are on the Histonet, I wish they would contact me so that I can include them in my workshops. Good luck and have fun with the technology, Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx -----Original Message----- From: Hallada, Teri [mailto:thallada@noch.org] Sent: Tuesday, October 28, 2003 10:43 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Microwave Tissue Processors I was wondering if anyone out there is using a microwave tissue processor for routine hospital tissues. Are there any regulations applicable to instituting one, ie FDA approval? Teri Hallada BS MT CT (ASCP) thallada@noch.org _______________________________________________ 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 have received this message in error, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From csawrenk <@t> bccancer.bc.ca Wed Oct 29 10:22:28 2003 From: csawrenk <@t> bccancer.bc.ca (Christina Sawrenko) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] Fli-1 Message-ID: <6BAF4D075F07D411B30900508B94CBA09C6D23@SERVER20> Good morning Histonetters, I am reposting a query regarding Fli-1 as my previous post did not produce any responses. We have now found a potential source for antibody: BC Pharmingen, cat#554267 and #554266. Does anyone have experience with these antibodies? We would want to use them on FFPE tissues for cases of Ewings's PNET. Thanks in advance for your help! Chris Sawrenko Histopatholology BC Cancer Agency Vancouver, BC, Canada From GAshton <@t> PICR.man.ac.uk Wed Oct 29 10:25:55 2003 From: GAshton <@t> PICR.man.ac.uk (Garry Ashton) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] freezing bones Message-ID: Dear all, After fixing and decalcifying some mouse bones, does anybody have experience at the best way to freeze the tissue down. Is simply a sucrose gradient, followed by snap freezing in liquid N2 adequate? Any advice and tips greatly appreciated. Garry Ashton Histology Dept 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. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031029/58b32907/attachment.htm From Sue.Kapoor <@t> uhsi.org Wed Oct 29 10:38:58 2003 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] alician yellow Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F54F4@khmcexch.uhsi.org> Hi all, I order alician yellow from NewComer Supply but am told that it's on backorder, can anyone suggest another vendor? Thanks in advance, Sue Kapoor, HT (ASCP) Kenosha Medical Center Kenosha, WI From Loralee_Gehan <@t> URMC.Rochester.edu Wed Oct 29 10:47:06 2003 From: Loralee_Gehan <@t> URMC.Rochester.edu (Gehan, Loralee) Date: Fri Sep 16 15:22:06 2005 Subject: [Histonet] re: purified water Message-ID: <95774A6A6036D411AFEA00D0B73C86430888046C@exmc3.urmc.rochester.edu> I would be careful with that. Our pathology department (special stains, histology and immunohistology) has been testing our distilled water through the microbiology lab for about one year now. Our distilled water has come back several times with bacteria in it. For any staining that we need to be careful of bacterial contaimination we use deionized. > ---------- > From: Suzannah B. Tieman > Reply To: tieman@albany.edu > Sent: Wednesday, October 29, 2003 5:19 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] re: purified water > > Hi all > In my experience, distilled water is more likely to be free of > bacteria. > Su Tieman > Suzannah B. Tieman > tieman@albany.edu > Biological Sciences, SUNY-Albany > 1400 Washington Ave > Albany, NY 12222 > ph: (518) 442-4317 > fax: (518) 442-4767 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From gudrun.lang <@t> aon.at Wed Oct 29 11:05:43 2003 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] When to use tap water? References: Message-ID: <008901c39e3e$e58ab6a0$0d12a8c0@SERVER> Hi I think distilled water was formerly a rather expensive reagens in laboratory. And so in each procedure, where Aqua dest. is not really necessary, the technicians prefer to use tap water. In my opinion tap water is necessary for blueing after hematoxylin and after the Schiff reagens , and tap water is forbidden with all silverstains before the fixation step. Otherwise there is no difference to rinse the slides with tap or dest. water. Gudrun Lang general hospital, Linz, Austria ----- Original Message ----- From: To: Sent: Wednesday, October 29, 2003 4:03 PM Subject: [Histonet] When to use tap water? > > Hello, all: > The prof for my graduate course in histology insisted that tap water be used for rinsing slides during certain staining procedures (eg. H&E)although he did not give a specific reason why. > When no "flavor" of water is specified in a procedure, I generally use MilliQ or distilled deionized water. Is this the safest route to take or should one use tap if nothing but "water" is listed? > Thanks for the attention. Maureen, MS candidate at ECU. > > -----Original Message----- > From: "Houston, Ronnie" <Ronnie_Houston@bshsi.com> > To: "'Morken, Tim - Labvision'" <tpmorken@labvision.com>, histonet@lists.utsouthwestern.edu > Sent: Tue, 28 Oct 2003 13:05:45 -0500 > Subject: RE: [Histonet] Dionized vs distilled water > > What quality of water is recommended/regulated for anatomic and clinical > pathology labs? > Ronnie Houston > Regional Histology Operations Manager > Bon Secours HealthPartners Laboratories > 5801 Bremo Road > Richmond, VA 23226 > (804) 287 7972 > ronnie_houston@bshsi.com > -----Original Message----- > From: Morken, Tim - Labvision [mailto:tpmorken@labvision.com] > Sent: Tuesday, October 28, 2003 12:53 PM > To: 'JCarpenter764@aol.com'; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Dionized vs distilled water > > > Distilled water is classically produced by heating water to evaporation > and subsequent condensing on a cold surface. In the process most > impurities are either evaporated off ahead of the water (in the case of > most organics), or left behind (in the case of minerals). The water is > also effectively deionized because the salts are left behind. It is > fairly pure water. To get very pure water it needs to be re-distilled > several times. > > Deionized water is classically passed through a salt bed or ionized > resin bed that captures the mineral ions (ie, a "water softener"). The > water is not necessarily pure, however, especially in regards to organic > chemicals. Reverse osmosis is also used now days to deionize water. > > > High quality water systems these days are some combination of filters, > distillation, deionizing resins and reverse osmosis. > > Tim Morken > > -----Original Message----- > From: JCarpenter764@aol.com [mailto:JCarpenter764@aol.com] > Sent: Tuesday, October 28, 2003 7:45 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] (no subject) > > while studying for my exam on the different fixatives and there > ingredients....i have noticed that some call for distilled water and > some use the term deionized water. Is there a difference? > > ________________________________________________________________________ > ________________________________________________________ > > ________________________________________________________________________ > ________________________________________________________ > > > > 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. > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From lao_ji <@t> yahoo.com Wed Oct 29 12:08:14 2003 From: lao_ji <@t> yahoo.com (Jimmy Lao) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] GFP antibodies In-Reply-To: <1E0CC447E59C974CA5C7160D2A2854EC459F40@SJMEMXMB04.stjude.sjcrh.local> Message-ID: <20031029180814.17670.qmail@web12508.mail.yahoo.com> Kristen, I think you are very close to success since you could see week signal in the Immunofluorescet staining. As my experience, you have to strickly keep all afterward process in Dark after the step you put fluorochrome conjugated Ab. My experiment is to use 1st Ab rabbit anti-GFP IgG fraction from Molecular Probes, the 2ndary Ab is Oregon Green goat anti-rabbit IgG(H+L). It works. even double staining with BrdU. Good Luck! Jimmy --- "Rothammer, Kristen" wrote: > Hello Everyone- > > > > I am desperately, and rather unsuccessfully, trying > to find a better way > to look at eGFP in frozen kidney sections from mice. > > I have read a wealth of info on histonet regarding > use of GFP > antibodies, and I have tried a number of methods, > but I am still not > having any luck. > > I have tried the following: > > > > (1) looking directly at eGFP in frozen sections > (sections are just a > few weeks old) right after 15 min fixation with 4% > paraformaldehyde > (fresh 16% paraformaldehyde diluted in PBS), > followed by washes in > PBS--> no GFP is observed > > (2) looking directly at eGFP in same frozen > sections without > fixation, coverslipping in PBS --> no GFP is > observed > > (3) fixation in paraformaldehyde as described > above followed by a > GFP antibody conjugated to a fluorochrome --> very > little signal is > observed > > (4) same fixation, followed by incubation with > GFP antibody, follwed > by biotinylated anti-rabbit, and finally by > streptavidin-Alexa 568 > > * This method resulted in EXTREMELY high > background. I tried to > block endogenous biotin by using both the egg/milk > protocol and > biotin/avidin drops from Vector Labs, both with > little success. > > > > I mount my slides with Flouromount, do NOT seal with > nail polish, and I > do not do antigen retrieval (this is not necessary > for frozen sections, > right?). > > > > > > Any input would be greatly appreciated! > > > > Thank you! > > > > Kristen Rothammer > > St. Jude Children's Research Hospital > > Department of Biochemistry > > Kristen.rothammer@stjude.org > > > > __________________________________ Do you Yahoo!? Exclusive Video Premiere - Britney Spears http://launch.yahoo.com/promos/britneyspears/ From Bauer.Karen <@t> mayo.edu Wed Oct 29 12:24:39 2003 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Not all tissue staining... Message-ID: <8C6E05FA69571948B461F1327CBB893E16C2C5@LMMAIL2> Hello to all, There are times when a stained IP slide will have only half of the tissue staining positively and not other parts of the tissue, but has the same cellularity of the part that is staining. (Did that make sense?) Same tissue cells, but some stain and some do not. When the Pathologist asks why this happens, I can only come up with possible vortex mixing problems or that the paraffin did not come out completely. We've recently switched to the BenchMark and this has only happened a few times. I've checked the vortex mixers, and all of the heating pads are working properly. Does anyone have any suggestions? Thanks in advance. Karen L. Bauer HT(ASCP) Histology Department Luther Hospital 715-838-3205 ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. From lchung <@t> ppmh.org Wed Oct 29 12:29:53 2003 From: lchung <@t> ppmh.org (Chung, Luong) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] anti-static/anti fatigue mats from Market Lab Message-ID: All, Has anyone use a anti-static mats from Market lab for Histology cutting area? Does it prevent static? Thank you for any feedback. Bruce Chung, MSM, CT(ASCP) Phoebe Putney Memorial Hospital Anatomic Pathology Manager HIPAA Privacy Rule requires covered entities to safeguard certain Protected Health Information (PHI) related to a person's healthcare. Information being sent to you may include PHI, after appropriate consent, acknowledgement, or authorization from the patient or under circumstances that do not require patient authorization. You, the recipient, are obligated to maintain PHI in a safe and secure manner. You may not re-disclose this patient information without additional patient consent or as required by law. Unauthorized re-disclosure or failure to safeguard PHI could subject us, or you, to penalties described in federal (HIPAA) and state law. If you, the reader of this message, are not the intended recipient, or the employee or agent responsible to deliver it to the intended recipient, please notify us immediately and destroy the related message. Thanks for your help. From laurie.colbert <@t> huntingtonhospital.com Wed Oct 29 12:47:26 2003 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Not all tissue staining... Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5BC60@EXCHANGE1.huntingtonhospital.com> Karen, We have four BenchMarks and and a NexES special stainer and whenever we have a problem we call Ventana's customer service. If they can't troubleshoot over the phone, they are very willing to send a service tech out. -----Original Message----- From: Bauer, Karen [mailto:Bauer.Karen@mayo.edu] Sent: Wednesday, October 29, 2003 10:25 AM To: Histonet (E-mail) Subject: [Histonet] Not all tissue staining... Hello to all, There are times when a stained IP slide will have only half of the tissue staining positively and not other parts of the tissue, but has the same cellularity of the part that is staining. (Did that make sense?) Same tissue cells, but some stain and some do not. When the Pathologist asks why this happens, I can only come up with possible vortex mixing problems or that the paraffin did not come out completely. We've recently switched to the BenchMark and this has only happened a few times. I've checked the vortex mixers, and all of the heating pads are working properly. Does anyone have any suggestions? Thanks in advance. Karen L. Bauer HT(ASCP) Histology Department Luther Hospital 715-838-3205 ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From david.kinsley <@t> spcorp.com Wed Oct 29 13:35:28 2003 From: david.kinsley <@t> spcorp.com (Kinsley, David) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] anti-static/anti fatigue mats from Market Lab Message-ID: I just spray the area with a little static guard and it work well. Dave -----Original Message----- From: Chung, Luong [mailto:lchung@ppmh.org] Sent: Wednesday, October 29, 2003 1:30 PM To: Histonet (E-mail) Subject: [Histonet] anti-static/anti fatigue mats from Market Lab All, Has anyone use a anti-static mats from Market lab for Histology cutting area? Does it prevent static? Thank you for any feedback. Bruce Chung, MSM, CT(ASCP) Phoebe Putney Memorial Hospital Anatomic Pathology Manager HIPAA Privacy Rule requires covered entities to safeguard certain Protected Health Information (PHI) related to a person's healthcare. Information being sent to you may include PHI, after appropriate consent, acknowledgement, or authorization from the patient or under circumstances that do not require patient authorization. You, the recipient, are obligated to maintain PHI in a safe and secure manner. You may not re-disclose this patient information without additional patient consent or as required by law. Unauthorized re-disclosure or failure to safeguard PHI could subject us, or you, to penalties described in federal (HIPAA) and state law. If you, the reader of this message, are not the intended recipient, or the employee or agent responsible to deliver it to the intended recipient, please notify us immediately and destroy the related message. Thanks for your help. _______________________________________________ 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 Patty.Lott <@t> ORTHO.UAB.EDU Wed Oct 29 14:09:25 2003 From: Patty.Lott <@t> ORTHO.UAB.EDU (Patty Lott) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] alkaline phosphatase stain on MMA sections Message-ID: <85F6C7A1330E794DB8540AFD001CC77E026F34@ROSCO> Has anyone tried alkaline phosphatase staining on MMA plastic sections of bone? I need a procedure, please! Patty Lott, Laboratory Supervisor Orthopaedic Research Laboratory Center for Metabolic Bone Disease Laboratory University of Alabama at Birmingham LHRB B37 0007 1919 7th Ave. South Birmingham, AL 35294 (205) 934-2007 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031029/9637f3e8/attachment.htm From mbryhan <@t> NORTHERNHEALTH.ORG Wed Oct 29 14:32:53 2003 From: mbryhan <@t> NORTHERNHEALTH.ORG (Mary Bryhan) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Prefer fixative Message-ID: Histonetters, I would like to set the record straight since I seemed to ruffle Hadi's feathers. Our institution does not perform FISH, but we do send them to Mayo Clinic in Rochester. We do our own immunohistochemistry and so far the only antibody that we have not been able to work up on Prefer fixed tissues is the TTF1. We have great results using Prefer fixative and I believe in this product. I believe that it does take an open mind, and a willingness on both staff and pathologists to work through the changes. Mary Bryhan HT (ASCP) Team Leader Anatomic Pathology Northern Michigan Hospital Petoskey, MI From emry <@t> u.washington.edu Wed Oct 29 14:34:49 2003 From: emry <@t> u.washington.edu (P. Emry) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] PBS Message-ID: Hi, I looked up Phospho Buffered Saline and got a few formulas I had not seen before. Is there a basic formula? Would you pass it on to me, please? Thanks for your help. Trisha From huffpw <@t> uleth.ca Wed Oct 29 15:23:33 2003 From: huffpw <@t> uleth.ca (Phillip Huff) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] PBS In-Reply-To: References: Message-ID: <1537.142.66.42.15.1067462613.squirrel@webmail.uleth.ca> The recipe that I have always used contains the following: 500 ?M KH2PO4, 5 mM Na2HPO4, 68.5 mM NaCl, 1350 ?M KCl, pH 7.4. Solutions are usually made up at 10x these concentrations and diluted to 1x prior to use. Don't forget to pH the solution at the temperature that you are going to use the solution as pH is temperature dependent. People often forget this minor fact and can affect your reaction, especially in cases such as ATPase reactions. Hope this helps, Phil Hi, > I looked up Phospho Buffered Saline and got a few formulas I had not seen > before. Is there a basic formula? Would you pass it on to me, please? > Thanks for your help. > > Trisha > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Kathy.Johnston <@t> CLS.ab.ca Wed Oct 29 15:30:08 2003 From: Kathy.Johnston <@t> CLS.ab.ca (Kathy.Johnston@CLS.ab.ca) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Black pigment on Bronch Lavages Message-ID: <30C050525B881C4AAFF41E6D16543E68015D02E1@mail3.cls.ab.ca> One of our pathologists and myself have been trying to identify some black intracellular pigmentation in a bronch lavage. We have ruled out carbon, and bleaching the section did not work, therefore is not melanin. It is a very fine dark black pigment and appears quite uniform in shape and size. Our pathologist is thinking that it is lead (the patient is a long time professional painter), but lead stains are negative. My other thought is aluminum deposits but have not yet stained for this. I am hoping someone on the "Net" may have some idea of what this may be, and if there is a method for demonstrating it. Thanks very much in advance! Kathy Johnston Tech II - Special Stains Anatomic Pathology - FMC Calgary Laboratory Services 1403-29 Street NW Calgary AB, Canada T2N 2T9 403-944-4760 403-290-4093 fax kathy.johnston@cls.ab.ca -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031029/7885b0c0/attachment.htm From nick.kirk3 <@t> btopenworld.com Wed Oct 29 16:08:40 2003 From: nick.kirk3 <@t> btopenworld.com (Nick Kirk) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] PBS In-Reply-To: Message-ID: There are numerous PBS recipes, most of which are determined by the pH you wish the buffer to be at. For Sorensen's Phosphate buffer you need two solutions:- A: 0.2M Sodium dihydrogen orthophosphate and B: 0.2M disodium hydrogen orthophosphate to make up the required pH solution you mix different volumes of each for the desired end pH. For example, pH 6.4 buffer requires 36.7 mls of A for every 13.3mls of B and pH 7.6 buffer requires 6.5mls of A for every 43.5 mls of B Most good Histology textbooks will have the recipes in them. It depends on which technique you are doing as to which buffer is required. Alternatively you can buy ready weighed "buffer tablets" that you just dissolve in distilled water. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of P. Emry Sent: 29 October 2003 20:35 To: HistoNet@Pathology.swmed.edu Subject: [Histonet] PBS Hi, I looked up Phospho Buffered Saline and got a few formulas I had not seen before. Is there a basic formula? Would you pass it on to me, please? Thanks for your help. Trisha _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mark.hayes <@t> mailbox.uq.edu.au Wed Oct 29 17:26:40 2003 From: mark.hayes <@t> mailbox.uq.edu.au (Mark Hayes) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Black pigment on Bronch Lavages In-Reply-To: <30C050525B881C4AAFF41E6D16543E68015D02E1@mail3.cls.ab.ca> Message-ID: HI Do you have access to an electron microscope? If so does it have an EDAX (if my memory serves me correctly) attachment or can you send it to one. I used one of these many years ago (circa 1982) at Sydney University to identify and semiquantitate some unique gold chloride crystals. EDAX or EDX stands for energy dispersive X-ray microanalysis (EDX). If you can isolate your specs onto an EM target you should at least be able to get an elemental analysis. Titanium oxide is also a component of paint and is utterly inert ? I think it needs HF to dissolve it. Hope this helps. Cheers MarkH -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Kathy.Johnston@CLS.ab.ca Sent: Thursday, 30 October 2003 7:30 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Black pigment on Bronch Lavages One of our pathologists and myself have been trying to identify some black intracellular pigmentation in a bronch lavage. We have ruled out carbon, and bleaching the section did not work, therefore is not melanin. It is a very fine dark black pigment and appears quite uniform in shape and size. Our pathologist is thinking that it is lead (the patient is a long time professional painter), but lead stains are negative. My other thought is aluminum deposits but have not yet stained for this. I am hoping someone on the "Net" may have some idea of what this may be, and if there is a method for demonstrating it. Thanks very much in advance! Kathy Johnston Tech II - Special Stains Anatomic Pathology - FMC Calgary Laboratory Services 1403-29 Street NW Calgary AB, Canada T2N 2T9 403-944-4760 403-290-4093 fax kathy.johnston@cls.ab.ca -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031030/594f92e9/attachment.htm From mark.hayes <@t> mailbox.uq.edu.au Wed Oct 29 17:34:25 2003 From: mark.hayes <@t> mailbox.uq.edu.au (Mark Hayes) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] antibodies for sheep - primers for common genes would also be good. Message-ID: HI all We have a number of sheep models in use in our research centre at the moment and are finding it difficult to get antibodies for immunohistochemistry. We would like to source antibodies which are specific for ovine proteins or are at lease crossreactive. We are also having difficulty getting gene sequences for common genes to generate primers. Can anyone help with sourcing either of these? Cheers MarkH Dr Mark Hayes BSc, MSc, Dip Ed (Tert), PhD Senior Lecturer, Department of Paediatrics and Child Health, University of Queensland Laboratory and Scientific Manager Royal Children's Hospital Foundation Research Centre L3 Foundation Building Royal Children's Hospital Herston Road, Herston Brisbane, Queensland. 4029 Ph 07 33655019 Fax 07 33655455 Mob 0402472024 email mark.hayes@uq.edu.au -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031030/3b4afc8c/attachment.htm From amarusk1 <@t> FAIRVIEW.ORG Wed Oct 29 18:07:47 2003 From: amarusk1 <@t> FAIRVIEW.ORG (ANN MARUSKA) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Uroplakin, WT1 Message-ID: Skipped content of type multipart/alternative-------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:ANN MARUSKA EMAIL;WORK;PREF;NGW:AMARUSK1.EAST.NORTH ORG:;LAB N:MARUSKA;ANN TEL;WORK:612-273-9119 END:VCARD From GREYTRUNK <@t> aol.com Wed Oct 29 18:15:17 2003 From: GREYTRUNK <@t> aol.com (GREYTRUNK@aol.com) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] histoplasma Message-ID: <1ce.136a9022.2cd1b215@aol.com> One of the techs that I work with got a weird request for an antibody to Histoplasma. Neither, she nor I have ever heard of one, byt she told me we would check it out. We have looked through the books we had available at work and came across nothing. Has anyone out there heard of an antibody for Histoplasma? If so, please let me know. Thanks Roxanne -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031029/15ecbdf7/attachment.htm From bryand <@t> netbistro.com Wed Oct 29 18:56:06 2003 From: bryand <@t> netbistro.com (Bryan Llewellyn) Date: Fri Sep 16 15:22:07 2005 Subject: Fw: [Histonet] Sirius Red Technique. Message-ID: <006701c39e81$00cfe420$8f70c2cf@bryand> ----- Original Message ----- From: Bryan Llewellyn To: Ian Montgomery Sent: Wednesday, October 29, 2003 9:48 AM Subject: Re: [Histonet] Sirius Red Technique. It very slowly deteriorates over time, but it can be compensated for by extending staining time. I have used solutions a year old with 2 hours staining, but that was stretching it a bit. Six months would be OK. So one hour fresh, 2 hours at six months. Incidentally, it is better if no more than 2 mL salt solution is added. The absolute minimum salt is the critical step. Bryan Llewellyn ----- Original Message ----- From: Ian Montgomery To: histonet@lists.utsouthwestern.edu Sent: Wednesday, October 29, 2003 2:46 AM Subject: [Histonet] Sirius Red Technique. Quick question and the man himself may be able to answer. Sirius Red Technique (Llewellyn, 1970), what is the shelf life of the prepared stain? 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 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031029/0100d605/attachment.htm From doscwk <@t> nus.edu.sg Wed Oct 29 21:02:00 2003 From: doscwk <@t> nus.edu.sg (Chan Wai Kam) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] RE: Microwave Tissue Processors Message-ID: Hi Mary, I was about to post a question to Histonet about the use of microwave for processing of tissues when I came across your message below. I hope to get some advice. I'm from Orthopaedics (research) and I process bone and cartilage specimens the usual way through fixation in formalin, then decalcification and so on. I'm just wondering whether I can use the microwave to speed up the processing without affecting the quality of the specimens. Our usual processing for bone takes around 3 weeks until embedding so it would be great if we could have something that can speed up the process without sacrifing quality. Would appreciate any advice out there. Thanks Julee Chan Orthopaedic Surgery National University of Singapore -----Original Message----- From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] Sent: Wednesday, October 29, 2003 11:38 PM To: ASelf@gmhsc.com Cc: histonet@lists.utsouthwestern.edu; Katherine Raker Subject: RE: [Histonet] RE: Microwave Tissue Processors Amy, Do you currently have a microwave tissue processor? We use the Hacker/Milestone RHS-1 Microwave to process our fatty tissue. The times that you see only represent the time at temperature, they do not include the "ramp" up time (the time needed to bring the solution to the appropriate processing temperature). Fixation - We fix the tissue cassettes overnight (6pm - 3am) in Alcoholic Zinc Formalin Ethanol - 10 minutes at 65 C JFC - Produced by Hacker. Isopropanol can be used instead, however, you may need to extend the time at temperature. - 1 hour and 37 minutes at 68 C Vaporization (Vacuum Drying) -Pressure at 500 mbar Paraffin - 48 minutes at 65 C and pressure at 100 mbar This program takes approximately 4 hours to run, it replaces our 16 hour overnight program that we previously used to process fatty tissue. The only fatty tissue we do not process in the microwave is lipoma. Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 >>> Amy Self 10/28/03 01:58PM >>> Mary, Could you share with me your procedure for processing fatty tissue in the microwave. Thanks, Amy Self -----Original Message----- From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] Sent: Tuesday, October 28, 2003 1:38 PM To: histonet@lists.utsouthwestern.edu; thallada@noch.org Subject: RE: [Histonet] RE: Microwave Tissue Processors We microwave process all of our biopsy specimens and our fatty tissue (except lipoma). Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 >>> "Hallada, Teri" 10/28/03 11:42AM >>> I was wondering if anyone out there is using a microwave tissue processor for routine hospital tissues. Are there any regulations applicable to instituting one, ie FDA approval? Teri Hallada BS MT CT (ASCP) thallada@noch.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or 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 hadi83 <@t> comcast.net Wed Oct 29 22:59:44 2003 From: hadi83 <@t> comcast.net (Hadi Yaziji) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Prefer fixative In-Reply-To: Message-ID: More on the Prefer issue: In a reference lab environment (like our lab), we have no choice in what our client-colleagues use to fix their specimens, so working with an open mind is not an option, but an obligation. I listed a few real problems that our techs (who have decades of experience with generating and validating antibodies) encounter with specimens fixed with Prefer. I also hear similar complaints from experienced colleagues during national advisory committee meetings. If Prefer is used purely to produce adequate H&E results, it is a good fixative. In terms of safety, I still look forward to long-term follow-up results about its safety. I have no romance with formalin, but any alternative fixative must be at least adequate for IHC and (the increasingly used) FISH. The Mayo works closely with Vysis, and Vysis/Abbott gives a long list of non-formalin fixatives (including Prefer) that makes it difficult to get good results using FISH. Finally, I am a pathologist, and I've learned a great deal from the wonderful experience of histotechs and Medtechs through this forum on technical issues, the same way I learn from techs in my workplace. On the other hand, I occasionally offer advice to the Histonet only when I feel it's useful to the group. Best, Hadi Yaziji, M.D. PhenoPath Laboratories On Wednesday, October 29, 2003, at 12:32 PM, Mary Bryhan wrote: > Histonetters, > > I would like to set the record straight since I seemed to ruffle > Hadi's feathers. > > Our institution does not perform FISH, but we do send them to Mayo > Clinic in Rochester. We do our own immunohistochemistry and so far > the only antibody that we have not been able to work up on Prefer > fixed tissues is the TTF1. > > We have great results using Prefer fixative and I believe in this > product. I believe that it does take an open mind, and a willingness > on both staff and pathologists to work through the changes. > > Mary Bryhan HT (ASCP) > Team Leader Anatomic Pathology > Northern Michigan Hospital > Petoskey, MI > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Kevin.Randall <@t> astrazeneca.com Thu Oct 30 02:58:34 2003 From: Kevin.Randall <@t> astrazeneca.com (Randall, Kevin J) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Cardboard slide trays uk Message-ID: <2429E934EAAAD511B6650001FA7E1A7D06D8C5@ukapphdevmsx04.ukapd.astrazeneca.net> Hello Histonetters, Can anybody out there give me a UK source for cardboard slide trays. We use the 24-slide, no flap variety. Our supplies are very nearly exhausted, I think our pathologists are using them to insulate their lofts! Thanks in advance Kevin Kevin Randall AstraZeneca Alderley Park Macclesfield Cheshire UK From c.m.vanderloos <@t> amc.uva.nl Thu Oct 30 04:19:10 2003 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] RE: When to use tap water? Message-ID: <680c79684a55.684a55680c79@amc.uva.nl> Hi all, There is an other very good reason to use tap water at some instances. After finishing your chromogen step at the end of an IHC staining, performed on an acetone-fixed cryostat section you better use tap water rather than distilled or MilliQ water. I found out that the nuclear morphology is very much destroyed doing so. Just 5 minutes in distilled water and your section looks like somebody just walked over it! Even a post-fixation step with buffered formaling prior to the distilled water rinse is not going to help you. Who is going to try this and confirm this funny result? Happy staining! Chris van der Loos Dept. of Cardiovascular Pathology Academical Medical Center Amsterdam - The Netherlands Original Message ----- >From Gary Gill Date Wed, 29 Oct 2003 10:26:10 -0500 To "'mrl0627@mail.ecu.edu'" , histonet@lists.utsouthwestern.edu Subject RE: [Histonet] When to use tap water? ------------------------------------------------------------------------ The pH of tap water is often higher than that for non-tap water, hence bluing of hematoxylin occurs slightly more quickly. Using "designer" water is overkill for staining applications, IMHO and experience. Gary Gill -----Original Message----- From: mrl0627@mail.ecu.edu [mailto:mrl0627@mail.ecu.edu] Sent: Wednesday, October 29, 2003 10:04 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] When to use tap water? Hello, all: The prof for my graduate course in histology insisted that tap water be used for rinsing slides during certain staining procedures (eg. H&E)although he did not give a specific reason why. When no "flavor" of water is specified in a procedure, I generally use MilliQ or distilled deionized water. Is this the safest route to take or should one use tap if nothing but "water" is listed? Thanks for the attention. Maureen, MS candidate at ECU. -----Original Message----- From: "Houston, Ronnie" To: "'Morken, Tim - Labvision'" , histonet@lists.utsouthwestern.edu Sent: Tue, 28 Oct 2003 13:05:45 -0500 Subject: RE: [Histonet] Dionized vs distilled water What quality of water is recommended/regulated for anatomic and clinical pathology labs? Ronnie Houston Regional Histology Operations Manager Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 ronnie_houston@bshsi.com -----Original Message----- From: Morken, Tim - Labvision [mailto:tpmorken@labvision.com] Sent: Tuesday, October 28, 2003 12:53 PM To: 'JCarpenter764@aol.com'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Dionized vs distilled water Distilled water is classically produced by heating water to evaporation and subsequent condensing on a cold surface. In the process most impurities are either evaporated off ahead of the water (in the case of most organics), or left behind (in the case of minerals). The water is also effectively deionized because the salts are left behind. It is fairly pure water. To get very pure water it needs to be re-distilled several times. Deionized water is classically passed through a salt bed or ionized resin bed that captures the mineral ions (ie, a "water softener"). The water is not necessarily pure, however, especially in regards to organic chemicals. Reverse osmosis is also used now days to deionize water. High quality water systems these days are some combination of filters, distillation, deionizing resins and reverse osmosis. Tim Morken -----Original Message----- From: JCarpenter764@aol.com [mailto:JCarpenter764@aol.com] Sent: Tuesday, October 28, 2003 7:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) while studying for my exam on the different fixatives and there ingredients....i have noticed that some call for distilled water and some use the term deionized water. Is there a difference? From lpwenk <@t> covad.net Thu Oct 30 04:24:12 2003 From: lpwenk <@t> covad.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Black pigment on Bronch Lavages References: <30C050525B881C4AAFF41E6D16543E68015D02E1@mail3.cls.ab.ca> Message-ID: <00e201c39ed0$0aeb8820$8732fea9@hppav> Just curious - how did you manage to "rule out carbon"? Since there isn't a histology special stain or a IHC antibody for carbon (that I'm aware of) - how was the possibility that it might be carbon "ruled out"? Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: Kathy.Johnston@CLS.ab.ca To: histonet@pathology.swmed.edu Sent: Wednesday, October 29, 2003 4:30 PM Subject: [Histonet] Black pigment on Bronch Lavages One of our pathologists and myself have been trying to identify some black intracellular pigmentation in a bronch lavage. We have ruled out carbon, and bleaching the section did not work, therefore is not melanin. It is a very fine dark black pigment and appears quite uniform in shape and size. Our pathologist is thinking that it is lead (the patient is a long time professional painter), but lead stains are negative. My other thought is aluminum deposits but have not yet stained for this. I am hoping someone on the "Net" may have some idea of what this may be, and if there is a method for demonstrating it. Thanks very much in advance! Kathy Johnston Tech II - Special Stains Anatomic Pathology - FMC Calgary Laboratory Services 1403-29 Street NW Calgary AB, Canada T2N 2T9 403-944-4760 403-290-4093 fax kathy.johnston@cls.ab.ca -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031030/3b605901/attachment.htm From ctsblack <@t> capeheart.uct.ac.za Thu Oct 30 05:07:23 2003 From: ctsblack <@t> capeheart.uct.ac.za (Melanie Black) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] T4000 or T3040 Message-ID: Hi There Does anybody in South Africa (or even better, in Cape Town) have any Technovit 4000 or Technovit 3040 adhesive, that you no longer need?? We would like to purchase it from you! Many Thanks Melanie Black University of Cape Town SA. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031030/ab075713/attachment.htm From ree3 <@t> leicester.ac.uk Thu Oct 30 05:26:09 2003 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] GST omega 1 Message-ID: Looking for a supplier for an antibody to the above..............many thanks.. Richard Edwards MRC TOXICOLOGY UNIT... LEICESTER....U.K....... From nick.kirk3 <@t> btopenworld.com Thu Oct 30 05:36:46 2003 From: nick.kirk3 <@t> btopenworld.com (nick.kirk3@btopenworld.com) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] GST omega 1 Message-ID: <3009665.1067513806457.JavaMail.root@127.0.0.1> Have you tried contacting Serotec? They offer a free antibody location service whereby they will find out suppliers (even if it's not themselves) for unusual or not so easily obtained antibodies. Try www.serotec.com for more info. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England > from: "Edwards, R.E." > date: Thu, 30 Oct 2003 11:26:09 > cc: histonet@lists.utsouthwestern.edu > subject: RE: [Histonet] GST omega 1 > > > > > > Looking for a supplier for an antibody to the above..............many thanks.. > Richard Edwards > MRC TOXICOLOGY UNIT... > LEICESTER....U.K....... > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jqb7 <@t> cdc.gov Thu Oct 30 06:02:32 2003 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] histoplasma Message-ID: The MSRS catalog of antibodies lists 4 companies with this antibody. Go to www.antibodies-probes.com and you can sign up and have all the info. at your fingertips. In the meantine, here are the 4 comapnies they listed: Meridian Diagnostics: (800) 543-1980 CSM, Inc.: (800) 241-0998 ImmunoMycologics, Inc.: (800) 654-3639 Universal Reagents: (317) 926-0006 Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: GREYTRUNK@aol.com [mailto:GREYTRUNK@aol.com] Sent: Wednesday, October 29, 2003 7:15 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] histoplasma One of the techs that I work with got a weird request for an antibody to Histoplasma. Neither, she nor I have ever heard of one, byt she told me we would check it out. We have looked through the books we had available at work and came across nothing. Has anyone out there heard of an antibody for Histoplasma? If so, please let me know. Thanks Roxanne -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031030/5c05934a/attachment.htm From Loralee_Gehan <@t> URMC.Rochester.edu Thu Oct 30 07:08:53 2003 From: Loralee_Gehan <@t> URMC.Rochester.edu (Gehan, Loralee) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] RE: Microwave Tissue Processors Message-ID: <95774A6A6036D411AFEA00D0B73C86430888046F@exmc3.urmc.rochester.edu> I work in an orthopaedics research lab as well and process many many many mouse hind limbs, calvaria, etc. We use a microwave processor from Hacker (milestone medical). It has decreased our time to process. We had some trouble at the start with some of our enzyme histochemistry. The trick to the machine is the heat. It was heating the samples up so much that one of our stains wasn't working. After much testing we figured out that all you have to do is decrease the temperature of the processing and increase the time. It was basically trial and error because the company markets these for rapid processing for surgical biopsies. We decal on the machine and we also do antigen retrieval and found that most of our antibodies worked well with it. We are still testing out the capabilities of the machine. But it has helped this lab become more efficient. Hope that helps. Loralee Gehan Orthopaedics Research Lab University of Rochester > ---------- > From: Chan Wai Kam > Sent: Wednesday, October 29, 2003 10:02 PM > To: Mary Reeves > Cc: HistoNet Server > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > Hi Mary, > > I was about to post a question to Histonet about the use of microwave > for processing of tissues when I came across your message below. I hope > to get some advice. > > I'm from Orthopaedics (research) and I process bone and cartilage > specimens the usual way through fixation in formalin, then > decalcification and so on. I'm just wondering whether I can use the > microwave to speed up the processing without affecting the quality of > the specimens. Our usual processing for bone takes around 3 weeks until > embedding so it would be great if we could have something that can speed > up the process without sacrifing quality. > > Would appreciate any advice out there. > > Thanks > Julee Chan > Orthopaedic Surgery > National University of Singapore > > > > -----Original Message----- > From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] > Sent: Wednesday, October 29, 2003 11:38 PM > To: ASelf@gmhsc.com > Cc: histonet@lists.utsouthwestern.edu; Katherine Raker > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > Amy, > Do you currently have a microwave tissue processor? We use the > Hacker/Milestone RHS-1 Microwave to process our fatty tissue. The times > that you see only represent the time at temperature, they do not include > the "ramp" up time (the time needed to bring the solution to the > appropriate processing temperature). > > Fixation - We fix the tissue cassettes overnight (6pm - 3am) in > Alcoholic Zinc Formalin Ethanol - 10 minutes at 65 C JFC - Produced by > Hacker. Isopropanol can be used instead, however, you may need to extend > the time at temperature. - 1 hour and 37 minutes at 68 C > Vaporization (Vacuum Drying) -Pressure at 500 mbar > Paraffin - 48 minutes at 65 C and pressure at 100 mbar > > This program takes approximately 4 hours to run, it replaces our 16 hour > overnight program that we previously used to process fatty tissue. The > only fatty tissue we do not process in the microwave is lipoma. > > Mary Reeves > Technical Specialist > Histology > 1-352-265-0680 ext 7-2113 > > >>> Amy Self 10/28/03 01:58PM >>> > > Mary, > > Could you share with me your procedure for processing fatty > tissue in the microwave. > > Thanks, > Amy Self > > -----Original Message----- > From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] > Sent: Tuesday, October 28, 2003 1:38 PM > To: histonet@lists.utsouthwestern.edu; thallada@noch.org > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > We microwave process all of our biopsy specimens and our fatty tissue > (except lipoma). > > Mary Reeves > Technical Specialist > Histology > 1-352-265-0680 ext 7-2113 > > >>> "Hallada, Teri" 10/28/03 11:42AM >>> > I was wondering if anyone out there is using a microwave tissue > processor for routine hospital tissues. Are there any regulations > applicable to instituting one, ie FDA approval? Teri Hallada BS MT CT > (ASCP) thallada@noch.org > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > Note: The information contained in this message may be privileged and > confidential and protected from disclosure. If the reader of this > message is not the intended recipient, or an employee or 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 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From mwhitesi <@t> adventisthealthcare.com Thu Oct 30 07:29:44 2003 From: mwhitesi <@t> adventisthealthcare.com (Marnie Whiteside) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Position in Maryland Message-ID: Hi, we have a position available for a supervisor in our histology lab. We're located in Takoma Park, Maryland. Any interested applicants can e-mail or call for further information. Marnie Whiteside Washington Adventist Hospital 301-891-5155/6102 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031030/c171f7f4/attachment.htm From Inga.Hansson <@t> neuro.uu.se Thu Oct 30 08:09:49 2003 From: Inga.Hansson <@t> neuro.uu.se (Inga Hansson) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] pressure cooking Message-ID: Hi all! I have an antibody that requires pressure cooking for 1 minute (after having reached temperature) but I have no pressure cooker. Is it possible to heat in microwave-owen instead and for how long? Inga -- Inga Hansson Dept. neuroscience, div. neurobiology PO Box 587, BMC Husargatan 3 S-751 23 Uppsala SWEDEN Phone: +46(18) 471 4384 Fax: +46(18)559017 From Rcartun <@t> harthosp.org Thu Oct 30 08:49:45 2003 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] histoplasma Message-ID: We do IHC for histoplasma. Richard Cartun >>> 10/29/03 07:15PM >>> One of the techs that I work with got a weird request for an antibody to Histoplasma. Neither, she nor I have ever heard of one, byt she told me we would check it out. We have looked through the books we had available at work and came across nothing. Has anyone out there heard of an antibody for Histoplasma? If so, please let me know. Thanks Roxanne From lesley <@t> vancouverbc.net Thu Oct 30 09:33:12 2003 From: lesley <@t> vancouverbc.net (Lesley Weston) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] re: purified water In-Reply-To: <95774A6A6036D411AFEA00D0B73C86430888046C@exmc3.urmc.rochester.edu> Message-ID: It might be a matter of how long the purified water is stored after purification and before use, rather than the method of purification. Lesley Weston. on 29/10/2003 8:47 AM, Gehan, Loralee at Loralee_Gehan@URMC.Rochester.edu wrote: > I would be careful with that. Our pathology department (special stains, > histology and immunohistology) has been testing our distilled water through > the microbiology lab for about one year now. Our distilled water has come > back several times with bacteria in it. For any staining that we need to be > careful of bacterial contaimination we use deionized. > >> ---------- >> From: Suzannah B. Tieman >> Reply To: tieman@albany.edu >> Sent: Wednesday, October 29, 2003 5:19 AM >> To: Histonet@lists.utsouthwestern.edu >> Subject: [Histonet] re: purified water >> >> Hi all >> In my experience, distilled water is more likely to be free of >> bacteria. >> Su Tieman >> Suzannah B. Tieman >> tieman@albany.edu >> Biological Sciences, SUNY-Albany >> 1400 Washington Ave >> Albany, NY 12222 >> ph: (518) 442-4317 >> fax: (518) 442-4767 >> >> _______________________________________________ >> 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 Thu Oct 30 09:36:23 2003 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Jennifer Hofecker Message-ID: I am trying to locate Jennifer Hofecker, formerly of the Univ. of Rochester Medical Center. If anyone knows of a way to contact her please reply. Thanks! Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031030/f79f0202/attachment.htm From Karen.Dresser <@t> umassmed.edu Thu Oct 30 10:02:33 2003 From: Karen.Dresser <@t> umassmed.edu (Dresser, Karen) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] histoplasma Message-ID: <5B238BE7511C7D4FB736CF0451643D530F80D3@edunivmail03.ad.umassmed.edu> Please note that "signing up" requires paying a subscription fee..... $98.00/year at least. Just FYI. Karen Dresser Research Histotechnician UMass Medical School -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: Thursday, October 30, 2003 7:03 AM To: 'GREYTRUNK@aol.com'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] histoplasma The MSRS catalog of antibodies lists 4 companies with this antibody. Go to www.antibodies-probes.com and you can sign up and have all the info. at your fingertips. In the meantine, here are the 4 comapnies they listed: Meridian Diagnostics: (800) 543-1980 CSM, Inc.: (800) 241-0998 ImmunoMycologics, Inc.: (800) 654-3639 Universal Reagents: (317) 926-0006 Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: GREYTRUNK@aol.com [mailto:GREYTRUNK@aol.com] Sent: Wednesday, October 29, 2003 7:15 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] histoplasma One of the techs that I work with got a weird request for an antibody to Histoplasma. Neither, she nor I have ever heard of one, byt she told me we would check it out. We have looked through the books we had available at work and came across nothing. Has anyone out there heard of an antibody for Histoplasma? If so, please let me know. Thanks Roxanne From jqb7 <@t> cdc.gov Thu Oct 30 10:03:41 2003 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] histoplasma Message-ID: You can also get a printed copy of the book if you don't want to subscribe to the on-line service. -----Original Message----- From: Dresser, Karen [mailto:Karen.Dresser@umassmed.edu] Sent: Thursday, October 30, 2003 11:03 AM To: Bartlett, Jeanine; GREYTRUNK@aol.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] histoplasma Please note that "signing up" requires paying a subscription fee..... $98.00/year at least. Just FYI. Karen Dresser Research Histotechnician UMass Medical School -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: Thursday, October 30, 2003 7:03 AM To: 'GREYTRUNK@aol.com'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] histoplasma The MSRS catalog of antibodies lists 4 companies with this antibody. Go to www.antibodies-probes.com and you can sign up and have all the info. at your fingertips. In the meantine, here are the 4 comapnies they listed: Meridian Diagnostics: (800) 543-1980 CSM, Inc.: (800) 241-0998 ImmunoMycologics, Inc.: (800) 654-3639 Universal Reagents: (317) 926-0006 Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: GREYTRUNK@aol.com [mailto:GREYTRUNK@aol.com] Sent: Wednesday, October 29, 2003 7:15 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] histoplasma One of the techs that I work with got a weird request for an antibody to Histoplasma. Neither, she nor I have ever heard of one, byt she told me we would check it out. We have looked through the books we had available at work and came across nothing. Has anyone out there heard of an antibody for Histoplasma? If so, please let me know. Thanks Roxanne From Karen.Dresser <@t> umassmed.edu Thu Oct 30 10:07:14 2003 From: Karen.Dresser <@t> umassmed.edu (Dresser, Karen) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] histoplasma Message-ID: <5B238BE7511C7D4FB736CF0451643D530F80D4@edunivmail03.ad.umassmed.edu> Actually, I just tried to order it, and the website says it is no longer available, apparently only the online subscription is available. -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: Thursday, October 30, 2003 11:04 AM To: Dresser, Karen; GREYTRUNK@aol.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] histoplasma You can also get a printed copy of the book if you don't want to subscribe to the on-line service. -----Original Message----- From: Dresser, Karen [mailto:Karen.Dresser@umassmed.edu] Sent: Thursday, October 30, 2003 11:03 AM To: Bartlett, Jeanine; GREYTRUNK@aol.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] histoplasma Please note that "signing up" requires paying a subscription fee..... $98.00/year at least. Just FYI. Karen Dresser Research Histotechnician UMass Medical School -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: Thursday, October 30, 2003 7:03 AM To: 'GREYTRUNK@aol.com'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] histoplasma The MSRS catalog of antibodies lists 4 companies with this antibody. Go to www.antibodies-probes.com and you can sign up and have all the info. at your fingertips. In the meantine, here are the 4 comapnies they listed: Meridian Diagnostics: (800) 543-1980 CSM, Inc.: (800) 241-0998 ImmunoMycologics, Inc.: (800) 654-3639 Universal Reagents: (317) 926-0006 Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: GREYTRUNK@aol.com [mailto:GREYTRUNK@aol.com] Sent: Wednesday, October 29, 2003 7:15 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] histoplasma One of the techs that I work with got a weird request for an antibody to Histoplasma. Neither, she nor I have ever heard of one, byt she told me we would check it out. We have looked through the books we had available at work and came across nothing. Has anyone out there heard of an antibody for Histoplasma? If so, please let me know. Thanks Roxanne From REEVEML <@t> shands.ufl.edu Thu Oct 30 10:13:00 2003 From: REEVEML <@t> shands.ufl.edu (Mary Reeves) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] RE: Microwave Tissue Processors Message-ID: Loralee, Which Hacker microwave do you have? We use the RHS-1 to process our decals. We are processing with vacuum, this allows us to process at lower temperatures and still have decreased processing times. We plan to start decalcification in the microwave soon. Any suggestions? Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 >>> "Gehan, Loralee" 10/30/03 08:08AM >>> I work in an orthopaedics research lab as well and process many many many mouse hind limbs, calvaria, etc. We use a microwave processor from Hacker (milestone medical). It has decreased our time to process. We had some trouble at the start with some of our enzyme histochemistry. The trick to the machine is the heat. It was heating the samples up so much that one of our stains wasn't working. After much testing we figured out that all you have to do is decrease the temperature of the processing and increase the time. It was basically trial and error because the company markets these for rapid processing for surgical biopsies. We decal on the machine and we also do antigen retrieval and found that most of our antibodies worked well with it. We are still testing out the capabilities of the machine. But it has helped this lab become more efficient. Hope that helps. Loralee Gehan Orthopaedics Research Lab University of Rochester > ---------- > From: Chan Wai Kam > Sent: Wednesday, October 29, 2003 10:02 PM > To: Mary Reeves > Cc: HistoNet Server > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > Hi Mary, > > I was about to post a question to Histonet about the use of microwave > for processing of tissues when I came across your message below. I hope > to get some advice. > > I'm from Orthopaedics (research) and I process bone and cartilage > specimens the usual way through fixation in formalin, then > decalcification and so on. I'm just wondering whether I can use the > microwave to speed up the processing without affecting the quality of > the specimens. Our usual processing for bone takes around 3 weeks until > embedding so it would be great if we could have something that can speed > up the process without sacrifing quality. > > Would appreciate any advice out there. > > Thanks > Julee Chan > Orthopaedic Surgery > National University of Singapore > > > > -----Original Message----- > From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] > Sent: Wednesday, October 29, 2003 11:38 PM > To: ASelf@gmhsc.com > Cc: histonet@lists.utsouthwestern.edu; Katherine Raker > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > Amy, > Do you currently have a microwave tissue processor? We use the > Hacker/Milestone RHS-1 Microwave to process our fatty tissue. The times > that you see only represent the time at temperature, they do not include > the "ramp" up time (the time needed to bring the solution to the > appropriate processing temperature). > > Fixation - We fix the tissue cassettes overnight (6pm - 3am) in > Alcoholic Zinc Formalin Ethanol - 10 minutes at 65 C JFC - Produced by > Hacker. Isopropanol can be used instead, however, you may need to extend > the time at temperature. - 1 hour and 37 minutes at 68 C > Vaporization (Vacuum Drying) -Pressure at 500 mbar > Paraffin - 48 minutes at 65 C and pressure at 100 mbar > > This program takes approximately 4 hours to run, it replaces our 16 hour > overnight program that we previously used to process fatty tissue. The > only fatty tissue we do not process in the microwave is lipoma. > > Mary Reeves > Technical Specialist > Histology > 1-352-265-0680 ext 7-2113 > > >>> Amy Self 10/28/03 01:58PM >>> > > Mary, > > Could you share with me your procedure for processing fatty > tissue in the microwave. > > Thanks, > Amy Self > > -----Original Message----- > From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] > Sent: Tuesday, October 28, 2003 1:38 PM > To: histonet@lists.utsouthwestern.edu; thallada@noch.org > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > We microwave process all of our biopsy specimens and our fatty tissue > (except lipoma). > > Mary Reeves > Technical Specialist > Histology > 1-352-265-0680 ext 7-2113 > > >>> "Hallada, Teri" 10/28/03 11:42AM >>> > I was wondering if anyone out there is using a microwave tissue > processor for routine hospital tissues. Are there any regulations > applicable to instituting one, ie FDA approval? Teri Hallada BS MT CT > (ASCP) thallada@noch.org > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > Note: The information contained in this message may be privileged and > confidential and protected from disclosure. If the reader of this > message is not the intended recipient, or an employee or 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 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From rcharles <@t> state.pa.us Thu Oct 30 10:14:33 2003 From: rcharles <@t> state.pa.us (Charles, Roger) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] ST5050 Message-ID: I am getting a variance of staining intensity on my 2 ST5050 autostainers. Has anyone else had this problem and can it be corrected? Roger Charles Microbiologist PA Department of Agriculture Veterinary Laboratory 2305 Cameron Street Harrisburg, PA 17110 717-787-8808 rcharles@state.pa.us From sharon.willman <@t> bms.com Thu Oct 30 10:30:30 2003 From: sharon.willman <@t> bms.com (Sharon E Willman) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] (Histonet) Tunel Kit and Fixative Message-ID: <3FA13CA6.40591426@bms.com> Hi, I was wondering if anyone has information on Tunel. What kit and fixative do you use? Has anyone used Bouin's? Any help you can give me would be most appreciated. Thanks, Sharon Willman From pmarcum <@t> polysciences.com Thu Oct 30 10:28:22 2003 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Jennifer Hofecker In-Reply-To: Message-ID: <000c01c39f02$d5af91f0$4800a8c0@PMARCUM2K> MessageI will tag on to Jeanine as I need Rhonda the Tennessee State President to contact me or tell me how to reach her. Please!!! Pam Marcum -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Bartlett, Jeanine Sent: Thursday, October 30, 2003 10:36 AM To: Histonet (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Jennifer Hofecker I am trying to locate Jennifer Hofecker, formerly of the Univ. of Rochester Medical Center. If anyone knows of a way to contact her please reply. Thanks! Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031030/f74442b3/attachment.htm From jqb7 <@t> cdc.gov Thu Oct 30 10:41:19 2003 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Jennifer Hofecker Message-ID: Thanks to everyone who gave me Jennifer's email address. Jeanine -----Original Message----- From: Pamela Marcum [mailto:pmarcum@polysciences.com] Sent: Thursday, October 30, 2003 11:28 AM To: Bartlett, Jeanine; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Jennifer Hofecker I will tag on to Jeanine as I need Rhonda the Tennessee State President to contact me or tell me how to reach her. Please!!! Pam Marcum -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Bartlett, Jeanine Sent: Thursday, October 30, 2003 10:36 AM To: Histonet (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Jennifer Hofecker I am trying to locate Jennifer Hofecker, formerly of the Univ. of Rochester Medical Center. If anyone knows of a way to contact her please reply. Thanks! Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031030/2a98c1ea/attachment.htm From Terry.Marshall <@t> rothgen.nhs.uk Thu Oct 30 10:31:07 2003 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Artifact quiz Message-ID: This was a new one on me. Go to http://pathcuric1.swmed.edu/histonet/index.htm and see if you can figure what this artifact was due to. 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 From DonnaWillis <@t> texashealth.org Thu Oct 30 11:11:36 2003 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] RE: Microwave Tissue Processors Message-ID: <5C6CBCCEB04B894C8BD15B312487F7B2205A88@ftwex01.txhealth.org> There is no true set time for decalfication in a microwave because bone type and section thickness and decal solutions vary. We have used Milestone, Energy Beam and TBS units with the same results. I prefer using a gentle formic acid solution because of IHC. The tissue must be fixed properly first then decal at 40C to 50C for 10 min. then check the sample. Bone Marrow Biopsies that are 1mm thick often don't need any longer than this. We have found that using a gentle formaic acid solution at 40C "usually" takes about 10 minutes for each mm of section thickness. Test it an see. The solution may need to be changed during the process if you are doing several cassettes or large tissues not in a cassette. Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx -----Original Message----- From: Chan Wai Kam [mailto:doscwk@nus.edu.sg] Sent: Wednesday, October 29, 2003 9:02 PM To: Mary Reeves Cc: HistoNet Server Subject: RE: [Histonet] RE: Microwave Tissue Processors Hi Mary, I was about to post a question to Histonet about the use of microwave for processing of tissues when I came across your message below. I hope to get some advice. I'm from Orthopaedics (research) and I process bone and cartilage specimens the usual way through fixation in formalin, then decalcification and so on. I'm just wondering whether I can use the microwave to speed up the processing without affecting the quality of the specimens. Our usual processing for bone takes around 3 weeks until embedding so it would be great if we could have something that can speed up the process without sacrifing quality. Would appreciate any advice out there. Thanks Julee Chan Orthopaedic Surgery National University of Singapore -----Original Message----- From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] Sent: Wednesday, October 29, 2003 11:38 PM To: ASelf@gmhsc.com Cc: histonet@lists.utsouthwestern.edu; Katherine Raker Subject: RE: [Histonet] RE: Microwave Tissue Processors Amy, Do you currently have a microwave tissue processor? We use the Hacker/Milestone RHS-1 Microwave to process our fatty tissue. The times that you see only represent the time at temperature, they do not include the "ramp" up time (the time needed to bring the solution to the appropriate processing temperature). Fixation - We fix the tissue cassettes overnight (6pm - 3am) in Alcoholic Zinc Formalin Ethanol - 10 minutes at 65 C JFC - Produced by Hacker. Isopropanol can be used instead, however, you may need to extend the time at temperature. - 1 hour and 37 minutes at 68 C Vaporization (Vacuum Drying) -Pressure at 500 mbar Paraffin - 48 minutes at 65 C and pressure at 100 mbar This program takes approximately 4 hours to run, it replaces our 16 hour overnight program that we previously used to process fatty tissue. The only fatty tissue we do not process in the microwave is lipoma. Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 >>> Amy Self 10/28/03 01:58PM >>> Mary, Could you share with me your procedure for processing fatty tissue in the microwave. Thanks, Amy Self -----Original Message----- From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] Sent: Tuesday, October 28, 2003 1:38 PM To: histonet@lists.utsouthwestern.edu; thallada@noch.org Subject: RE: [Histonet] RE: Microwave Tissue Processors We microwave process all of our biopsy specimens and our fatty tissue (except lipoma). Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 >>> "Hallada, Teri" 10/28/03 11:42AM >>> I was wondering if anyone out there is using a microwave tissue processor for routine hospital tissues. Are there any regulations applicable to instituting one, ie FDA approval? Teri Hallada BS MT CT (ASCP) thallada@noch.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or 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 _______________________________________________ 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 have received this message in error, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From Denise.Griffiths <@t> genzyme.com Thu Oct 30 10:44:55 2003 From: Denise.Griffiths <@t> genzyme.com (Griffiths, Denise) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] unsubscribe Message-ID: <20309DC1E4E8D6119619000255DB08BB3CCD61@MMAIL1> -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031030/45372ffe/attachment.htm From tvedilago <@t> system1.net Thu Oct 30 11:52:10 2003 From: tvedilago <@t> system1.net (Tommy Vedilago) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Histology Jobs Message-ID: Hello Histonetters, We are currently recruiting for many positions around the country. We have Supervisor/Management positions in New York City, Philadelphia, New Jersey, and Tampa FL. We also are looking for techs for Ft Lauderdale, Nashville, Michigan, Atlanta, Dallas, and Northern New Jersey. I look forward to hearing from all of you. I am also looking for techs and supervisors for traveling or temporary positions as well. We expect to begin those placements in the next 30 days or so. Please feel free to contact me at any time at 866-SYSTEM1 or 866-797-8361. Thanks, Tommy Vedilago System 1 Search (864) 627-0012 (864) 627-0013 Fax From Rcartun <@t> harthosp.org Thu Oct 30 12:31:36 2003 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] CA125 in Breast CA Message-ID: Has anyone seen diffuse, strong immunoreactivity for CA125 in a high grade ductal breast CA? Thanks! Richard Cartun From lao_ji <@t> yahoo.com Thu Oct 30 13:27:14 2003 From: lao_ji <@t> yahoo.com (Jimmy Lao) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] (Histonet) Tunel Kit and Fixative In-Reply-To: <3FA13CA6.40591426@bms.com> Message-ID: <20031030192714.34451.qmail@web12505.mail.yahoo.com> Sharon, I use the "ApopTag Fluorescein I situ Apoptosis Detection Kit" from Serologicals Corporation Catalog#:S7110 Maybe it has other company name: Intergen Company. The one I used is anti-Dig conjugated rhodamine. the disadvantage is that you need to distinguish the deed cells from the autofluoresein cells like some blood cell. maybe selecting fluoresein as the conjugating fluorochrome is better. Jimmy --- Sharon E Willman wrote: > Hi, > I was wondering if anyone has information on Tunel. > What kit > and fixative do you use? Has anyone used Bouin's? > Any help you > can give me would be most appreciated. > > Thanks, > > Sharon Willman > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________ Do you Yahoo!? Exclusive Video Premiere - Britney Spears http://launch.yahoo.com/promos/britneyspears/ From burch007 <@t> mc.duke.edu Thu Oct 30 14:20:18 2003 From: burch007 <@t> mc.duke.edu (James L Burchette) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Histochem reagent Ethopropazine Message-ID: Hello All: Does anyone have a source for Ethopropazine used in the acetylcholinesterase histochemistry. The product has discontinued by Sigma. Fisher does not offer it either. Thanks, Jim B -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031030/b16290e0/attachment.htm From Loralee_Gehan <@t> URMC.Rochester.edu Thu Oct 30 14:30:08 2003 From: Loralee_Gehan <@t> URMC.Rochester.edu (Gehan, Loralee) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] RE: Microwave Tissue Processors Message-ID: <95774A6A6036D411AFEA00D0B73C864308880471@exmc3.urmc.rochester.edu> We have the RHS-1 also. What we normally do for our decals is to let them sit in EDTA for about a week sometimes longer depending on the amount of things that we have to cut/process, etc. Then we run it on the decal cycle in fresh EDTA at 37C for about 5 hours then process them the next day. We have been getting beautiful results. Do you do your entire processing schedule under vacuum? Or just the paraffin? Loralee > ---------- > From: Mary Reeves > Sent: Thursday, October 30, 2003 11:13 AM > To: doscwk@nus.edu.sg; Loralee_Gehan@urmc.rochester.edu > Cc: histonet@lists.utsouthwestern.edu; Katherine Raker > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > Loralee, > Which Hacker microwave do you have? We use the RHS-1 to process our > decals. We are processing with vacuum, this allows us to process at lower > temperatures and still have decreased processing times. We plan to start > decalcification in the microwave soon. Any suggestions? > > Mary Reeves > Technical Specialist > Histology > 1-352-265-0680 ext 7-2113 > > >>> "Gehan, Loralee" 10/30/03 08:08AM > >>> > I work in an orthopaedics research lab as well and process many many many > mouse hind limbs, calvaria, etc. We use a microwave processor from Hacker > (milestone medical). It has decreased our time to process. We had some > trouble at the start with some of our enzyme histochemistry. > The trick to the machine is the heat. It was heating the samples up so > much > that one of our stains wasn't working. After much testing we figured out > that all you have to do is decrease the temperature of the processing and > increase the time. It was basically trial and error because the company > markets these for rapid processing for surgical biopsies. > We decal on the machine and we also do antigen retrieval and found that > most > of our antibodies worked well with it. We are still testing out the > capabilities of the machine. But it has helped this lab become more > efficient. > Hope that helps. > > Loralee Gehan > Orthopaedics Research Lab > University of Rochester > > ---------- > > From: Chan Wai Kam > > Sent: Wednesday, October 29, 2003 10:02 PM > > To: Mary Reeves > > Cc: HistoNet Server > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > Hi Mary, > > > > I was about to post a question to Histonet about the use of microwave > > for processing of tissues when I came across your message below. I hope > > to get some advice. > > > > I'm from Orthopaedics (research) and I process bone and cartilage > > specimens the usual way through fixation in formalin, then > > decalcification and so on. I'm just wondering whether I can use the > > microwave to speed up the processing without affecting the quality of > > the specimens. Our usual processing for bone takes around 3 weeks until > > embedding so it would be great if we could have something that can speed > > up the process without sacrifing quality. > > > > Would appreciate any advice out there. > > > > Thanks > > Julee Chan > > Orthopaedic Surgery > > National University of Singapore > > > > > > > > -----Original Message----- > > From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] > > Sent: Wednesday, October 29, 2003 11:38 PM > > To: ASelf@gmhsc.com > > Cc: histonet@lists.utsouthwestern.edu; Katherine Raker > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > > > Amy, > > Do you currently have a microwave tissue processor? We use the > > Hacker/Milestone RHS-1 Microwave to process our fatty tissue. The times > > that you see only represent the time at temperature, they do not include > > the "ramp" up time (the time needed to bring the solution to the > > appropriate processing temperature). > > > > Fixation - We fix the tissue cassettes overnight (6pm - 3am) in > > Alcoholic Zinc Formalin Ethanol - 10 minutes at 65 C JFC - Produced by > > Hacker. Isopropanol can be used instead, however, you may need to extend > > the time at temperature. - 1 hour and 37 minutes at 68 C > > Vaporization (Vacuum Drying) -Pressure at 500 mbar > > Paraffin - 48 minutes at 65 C and pressure at 100 mbar > > > > This program takes approximately 4 hours to run, it replaces our 16 hour > > overnight program that we previously used to process fatty tissue. The > > only fatty tissue we do not process in the microwave is lipoma. > > > > Mary Reeves > > Technical Specialist > > Histology > > 1-352-265-0680 ext 7-2113 > > > > >>> Amy Self 10/28/03 01:58PM >>> > > > > Mary, > > > > Could you share with me your procedure for processing fatty > > tissue in the microwave. > > > > Thanks, > > Amy Self > > > > -----Original Message----- > > From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] > > Sent: Tuesday, October 28, 2003 1:38 PM > > To: histonet@lists.utsouthwestern.edu; thallada@noch.org > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > > > We microwave process all of our biopsy specimens and our fatty tissue > > (except lipoma). > > > > Mary Reeves > > Technical Specialist > > Histology > > 1-352-265-0680 ext 7-2113 > > > > >>> "Hallada, Teri" 10/28/03 11:42AM >>> > > I was wondering if anyone out there is using a microwave tissue > > processor for routine hospital tissues. Are there any regulations > > applicable to instituting one, ie FDA approval? Teri Hallada BS MT CT > > (ASCP) thallada@noch.org > > > > > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > Note: The information contained in this message may be privileged and > > confidential and protected from disclosure. If the reader of this > > message is not the intended recipient, or an employee or 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 > > > > _______________________________________________ > > 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 Oct 30 14:34:55 2003 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] simulated body fluid Message-ID: <91.34fe2a92.2cd2cfef@aol.com> Skipped content of type multipart/alternative-------------- next part -------------- A non-text attachment was scrubbed... Name: not available Type: image/jpeg Size: 2082 bytes Desc: not available Url : http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031030/9ee8a32a/attachment.jpg From Kathy.Johnston <@t> CLS.ab.ca Thu Oct 30 15:02:46 2003 From: Kathy.Johnston <@t> CLS.ab.ca (Kathy.Johnston@CLS.ab.ca) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Black pigment on Bronch Lavages Message-ID: <30C050525B881C4AAFF41E6D16543E68015D02E4@mail3.cls.ab.ca> As our pathologist was explaining his problem to me while we looked at the slide in particular under his microscope, he said that Carbon was not the item in question as it does not become refractile when he polarizes/darkfield his microscope (to be honest he was whizzing objectives and filters so fast I could barely follow him!). I know carbon is very common in lung, but to my eye as well, it is too fine and regular to resemble the stuff I usually see. -----Original Message----- From: thehud@ldd.net [mailto:thehud@ldd.net] Sent: Wednesday, October 29, 2003 3:31 PM To: Kathy.Johnston@CLS.ab.ca Subject: Re: [Histonet] Black pigment on Bronch Lavages dear kathy, how have you ruled out carbon, as this is so common. peter h. dohan, md -----Original Message----- From: Kathy.Johnston@CLS.ab.ca Sent: Oct 29, 2003 1:30 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Black pigment on Bronch Lavages One of our pathologists and myself have been trying to identify some black intracellular pigmentation in a bronch lavage. We have ruled out carbon, and bleaching the section did not work, therefore is not melanin. It is a very fine dark black pigment and appears quite uniform in shape and size. Our pathologist is thinking that it is lead (the patient is a long time professional painter), but lead stains are negative. My other thought is aluminum deposits but have not yet stained for this. I am hoping someone on the "Net" may have some idea of what this may be, and if there is a method for demonstrating it. Thanks very much in advance! Kathy Johnston Tech II - Special Stains Anatomic Pathology - FMC Calgary Laboratory Services 1403-29 Street NW Calgary AB, Canada T2N 2T9 403-944-4760 403-290-4093 fax kathy.johnston@cls.ab.ca -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031030/cd80fa3c/attachment.htm From Don.Birgerson <@t> leica-microsystems.com Thu Oct 30 15:18:32 2003 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] ST5050 Message-ID: Roger, Your message mentioned an Autostainer, ST5050. The ST5050 is the "Histostainer Ig" while the Autostainer is ST5010. Give me a call and I'll try to help. Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Charles, Roger" To: Sent by: cc: histonet-admin@lists.utsouth Subject: [Histonet] ST5050 western.edu 10/30/2003 10:14 AM I am getting a variance of staining intensity on my 2 ST5050 autostainers. Has anyone else had this problem and can it be corrected? Roger Charles Microbiologist PA Department of Agriculture Veterinary Laboratory 2305 Cameron Street Harrisburg, PA 17110 717-787-8808 rcharles@state.pa.us _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________ This email has been scanned for all viruses by the MessageLabs Email Security System. For more information on a proactive email security service working around the clock, around the globe, visit http://www.messagelabs.com ________________________________________________________________________ From amosbrooks <@t> earthlink.net Thu Oct 30 15:10:40 2003 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] No lab is an island In-Reply-To: <20031029180000.16294.8742.Mailman@swlx167.swmed.edu> References: <20031029180000.16294.8742.Mailman@swlx167.swmed.edu> Message-ID: <6.0.0.22.0.20031030155027.029b43e0@127.0.0.1> Mary, I am sure you can get acceptable results using Prefer. The problem is that it is not standardized. The manufacturers of antibodies test them on formalin fixed tissues or frozen sections. Those are the choices. If you use something else then the results you get from the tests you do is limited to your lab only. If you send tissue out, other labs can not perform the same tests or more specifically many of the tests not done at your facility. Perhaps manufacturers should look into testing other fixatives. The problem here is that there are so many fixatives on the market right now that unless there was some collusion (which would be unethical) between the companies they can't pick any one company with out alienating all the others. If we want to change fixatives from formalin to something else, we need as a WHOLE to establish what EVERY lab MUST switch to and start from there. (Perhaps some regulatory agency would be of some help there ... ouch) Because if we do not then there will be pockets of Prefer and pockets and PenFix and pockets of formalin. With out an established standard the labs can not communicate. There would need to be countless procedures and controls fixed in countless methods. Imagine trying to find an ALK-1 control fixed 10 different ways. (Good bloody luck, it's hard enough finding it in formalin). Prefer may indeed be better but currently, for better or for worse, we are stuck with formalin since it is the standard. Amos Brooks At 01:00 PM 10/29/03, you wrote: >Message: 11 >Date: Wed, 29 Oct 2003 09:18:38 -0500 >From: "Mary Bryhan" >To: , >Cc: >Subject: Re: [Histonet] Prefer formalin-free fixative > >All, > >I would like to go on record to dispute the claims of Hadi regarding = >Prefer fixative. It is much safer than formalin based fixatives for the = >health of the workers as well as the environment. The main reason people = >have trouble with working up their antibodies using Prefer fixed tissues = >are, in most cases, they haven't tried to work through the protocols to = >determine the corrective action to get their signal to light up! We have = >had about 8 years or so, experience with using Prefer fixative and have = >EXCELLENT results! If anyone out there needs to network with other Prefer = >users, feel free to contact myself or my coworker Ellen Clausen-Simon at = >(231)487-4169. > >Mary Bryhan HT (ASCP) >Team Leader Anatomic Pathology Department >Northern Michigan Hospital=20 >Petoskey, MI 49770 From tpmorken <@t> labvision.com Thu Oct 30 15:50:31 2003 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] No lab is an island Message-ID: <8CD0CDC791DCC343ABB1F15298E4F3C417CC88@usca0082k03.rallansci.apogent.com> Amos brings up some good points about IHC with various fixtative. Another problem with all these different fixatives and IHC is that antibody vendors have a hard time getting even formalin-fixed tissue blocks as it is, and trying to get blocks of tissue treated with many different fixatives would be almost impossible, or at least extremely expensive. Also, antibody vendors have invested huge amounst of money to develop antibodies for formalin-fixed tissue simply because it is the standard. I think it is good that people try other fixatives, but only publishing superior results will convince people to change. Tim Morken Product Development Lab Vision / NeoMarkers 47790 Westinghouse Dr Fremont, CA 94539 PH: 510-991-2840 www.labvision.com -----Original Message----- From: Amos Brooks [mailto:amosbrooks@earthlink.net] Sent: Thursday, October 30, 2003 1:11 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] No lab is an island Mary, I am sure you can get acceptable results using Prefer. The problem is that it is not standardized. The manufacturers of antibodies test them on formalin fixed tissues or frozen sections. Those are the choices. If you use something else then the results you get from the tests you do is limited to your lab only. If you send tissue out, other labs can not perform the same tests or more specifically many of the tests not done at your facility. Perhaps manufacturers should look into testing other fixatives. The problem here is that there are so many fixatives on the market right now that unless there was some collusion (which would be unethical) between the companies they can't pick any one company with out alienating all the others. If we want to change fixatives from formalin to something else, we need as a WHOLE to establish what EVERY lab MUST switch to and start from there. (Perhaps some regulatory agency would be of some help there ... ouch) Because if we do not then there will be pockets of Prefer and pockets and PenFix and pockets of formalin. With out an established standard the labs can not communicate. There would need to be countless procedures and controls fixed in countless methods. Imagine trying to find an ALK-1 control fixed 10 different ways. (Good bloody luck, it's hard enough finding it in formalin). Prefer may indeed be better but currently, for better or for worse, we are stuck with formalin since it is the standard. Amos Brooks At 01:00 PM 10/29/03, you wrote: >Message: 11 >Date: Wed, 29 Oct 2003 09:18:38 -0500 >From: "Mary Bryhan" >To: , >Cc: >Subject: Re: [Histonet] Prefer formalin-free fixative > >All, > >I would like to go on record to dispute the claims of Hadi regarding = >Prefer fixative. It is much safer than formalin based fixatives for the = >health of the workers as well as the environment. The main reason people = >have trouble with working up their antibodies using Prefer fixed tissues = >are, in most cases, they haven't tried to work through the protocols to = >determine the corrective action to get their signal to light up! We have = >had about 8 years or so, experience with using Prefer fixative and have = >EXCELLENT results! If anyone out there needs to network with other Prefer = >users, feel free to contact myself or my coworker Ellen Clausen-Simon at = >(231)487-4169. > >Mary Bryhan HT (ASCP) >Team Leader Anatomic Pathology Department >Northern Michigan Hospital=20 >Petoskey, MI 49770 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BRBaca <@t> salud.unm.edu Thu Oct 30 15:51:23 2003 From: BRBaca <@t> salud.unm.edu (Brandy Baca) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] liquid nitrogen immersion Message-ID: I am new to the listserve and have a question regarding immersion of cassettes and vials in liquid nitrogen. I am going to be working with a developing tissue bank and wondered if anyone knows what cassettes and vials can withstand immersion in liquid nitrogen for quick-freezing. I have spoke with Sakura and they have no evidence that their uni-cassettes can withstand immersion. As for vials, many don't reccomend immersion but some require a tube in order to accomodate this process. If anyone can suggest a manufacturer and type, it would be greatly appreciated. I would like to thank everyone in advance for their responses. Brandy Baca Dept of Pathology CRTC B-93 505.272.1127 phone 505.272.1370 fax From gareth.davis <@t> Vanderbilt.Edu Thu Oct 30 16:06:54 2003 From: gareth.davis <@t> Vanderbilt.Edu (Davis, Gareth) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] liquid nitrogen immersion Message-ID: <082C721AF78DB34983E8BA2CD08546216A40DB@mailbe07> Brandy, When we have immersed tissue into Liquid Nitrogen, we have actually made our own cassettes with foil. We just use regular foil and shape it around a cork or bottom of a small bottle (depending on size needed), leaving extra to fold over the frozen block - when finished freezing. Put the freezing medium and tissue in foil cassette and immerse in LN until just about completely frozen, =small bubble in center results. Then remove and store as in -80. Hope this helps. Gareth Ms. Gareth B. Davis Research Assistant II Neuro-magnetics Division of the Department of Neurology Vanderbilt University Medical Center 615-936-3318 -----Original Message----- From: Brandy Baca [mailto:BRBaca@salud.unm.edu] Sent: Thu 10/30/2003 3:51 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] liquid nitrogen immersion I am new to the listserve and have a question regarding immersion of cassettes and vials in liquid nitrogen. I am going to be working with a developing tissue bank and wondered if anyone knows what cassettes and vials can withstand immersion in liquid nitrogen for quick-freezing. I have spoke with Sakura and they have no evidence that their uni-cassettes can withstand immersion. As for vials, many don't reccomend immersion but some require a tube in order to accomodate this process. If anyone can suggest a manufacturer and type, it would be greatly appreciated. I would like to thank everyone in advance for their responses. Brandy Baca Dept of Pathology CRTC B-93 505.272.1127 phone 505.272.1370 fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031030/920062c8/attachment.htm From peoshel <@t> wisc.edu Thu Oct 30 16:07:04 2003 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] liquid nitrogen immersion In-Reply-To: References: Message-ID: Brandy, Most scientific supply companies carry cryovials, but these may not be what you need. We store samples in ordinary minifuge tubes, which are made of polypropylene. There are also larger vials for storage in LN2 using "sticks" to hold the vials in the storage dewars. They are also available from Fisher and so forth. I wouldn't try using standard histology specimen cassettes, though, simply because they're designed for specimen processing in different chemicals and raised temperatures. If there's a sperm bank near you, ask what they use, or check the folks in molecular biology, etc., at the University. Phil >I am new to the listserve and have a question regarding immersion of >cassettes and vials in liquid nitrogen. I am going to be working with a >developing tissue bank and wondered if anyone knows what cassettes and >vials can withstand immersion in liquid nitrogen for quick-freezing. I >have spoke with Sakura and they have no evidence that their >uni-cassettes can withstand immersion. As for vials, many don't >reccomend immersion but some require a tube in order to accomodate this >process. If anyone can suggest a manufacturer and type, it would be >greatly appreciated. I would like to thank everyone in advance for their >responses. > > > >Brandy Baca >Dept of Pathology >CRTC B-93 >505.272.1127 phone >505.272.1370 fax > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From algranth <@t> u.arizona.edu Thu Oct 30 16:08:08 2003 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Sirius red Message-ID: <4.3.2.7.2.20031030145119.00ccb050@algranth.inbox.email.arizona.edu> I have a quick question about Sirius red since it is a recent topic of discussion on histonet. I was asked to do the stain and given a protocol calling for Sirius red F3B (CI 35782). I have Sirius red CI 35780. They said that they have tried the stain using CI 35780 and it didn't work but they cannot find the other Sirius red with a CI# 35782 in any of the catalogs. We have tried Sigma, Polysciences, PolyScientific, Newcomer, and some others... Any suggestions where I can find it? Andi Grantham ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From bhewlett <@t> cogeco.ca Thu Oct 30 16:49:02 2003 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Sirius red References: <4.3.2.7.2.20031030145119.00ccb050@algranth.inbox.email.arizona.edu> Message-ID: <006e01c39f38$06880bb0$6400a8c0@bryaniwx13voft> Andrea, The dye that you have - CI # 35780 is Sirius red F3B (CI Direct red 80), this is the correct dye for collagen staining. I would suggest that you use this, just because 'they' tried the stain and it didn't work doesn't mean it doesn't! I suspect that you will be unable to find the mythical CI 35782. Bryan ----- Original Message ----- From: "Andrea Grantham" To: Sent: Thursday, October 30, 2003 5:08 PM Subject: [Histonet] Sirius red > I have a quick question about Sirius red since it is a recent topic of > discussion on histonet. > I was asked to do the stain and given a protocol calling for Sirius red F3B > (CI 35782). I have Sirius red CI 35780. They said that they have tried the > stain using CI 35780 and it didn't work but they cannot find the other > Sirius red with a CI# 35782 in any of the catalogs. We have tried Sigma, > Polysciences, PolyScientific, Newcomer, and some others... > Any suggestions where I can find it? > Andi Grantham > ..................................................................... > : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > : Sr. Research Specialist University of Arizona : > : (office: AHSC 4212) P.O. Box 245044 : > : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : > :...................................................................: > http://www.cba.arizona.edu/histology-lab.html > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jnocito <@t> satx.rr.com Thu Oct 30 19:45:36 2003 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Cutting Bee heads Message-ID: <000b01c39f50$aec741c0$70494542@satx.rr.com> Hello all, I'm helping this high school student with his science project. He needs bee's heads cut to include a complete cross section of brain. Since all my experience is clinical, what is the best way to cut through the exoskeleton? I've processed the heads and tried using Nair to soften up the keratin, but is there another way to process these things. Thanks for your help. Joe Nocito BS, HT (ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, Texas From mark.hayes <@t> mailbox.uq.edu.au Thu Oct 30 18:33:48 2003 From: mark.hayes <@t> mailbox.uq.edu.au (Mark Hayes) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] antibodies for sheep - primers for common genes would also be good. In-Reply-To: Message-ID: Hi Albert Thanks for the reply. For starters we would love reliable antibodies to IL6, 8, 10; TNFa, TGFB2&3, Fibronectin, Collagens 1&3, FGFs, FGFRs, BMPs and COX-2. There are quite a few others we would like but as you can see the list is rather extensive/expensive already. Cheers MarkH -----Original Message----- From: Albert Grobe [mailto:GrobeA@saintpatrick.org] Sent: Friday, 31 October 2003 1:52 AM To: mark.hayes@uq.edu.au Subject: Re: [Histonet] antibodies for sheep - primers for common genes would also be good. What proteins/epitopes are you looking for? Our lab works with sheep as well and we have several abs that work well. Albert >>> "Mark Hayes" 10/29/03 04:34PM >>> HI all We have a number of sheep models in use in our research centre at the moment and are finding it difficult to get antibodies for immunohistochemistry. We would like to source antibodies which are specific for ovine proteins or are at lease crossreactive. We are also having difficulty getting gene sequences for common genes to generate primers. Can anyone help with sourcing either of these? Cheers MarkH Dr Mark Hayes BSc, MSc, Dip Ed (Tert), PhD Senior Lecturer, Department of Paediatrics and Child Health, University of Queensland Laboratory and Scientific Manager Royal Children's Hospital Foundation Research Centre L3 Foundation Building Royal Children's Hospital Herston Road, Herston Brisbane, Queensland. 4029 Ph 07 33655019 Fax 07 33655455 Mob 0402472024 email mark.hayes@uq.edu.au From katri <@t> cogeco.ca Thu Oct 30 19:09:44 2003 From: katri <@t> cogeco.ca ( Katri Tuomala) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] pressure cooking References: Message-ID: <00a201c39f4b$ad568d00$ce989618@hala4.on.cogeco.ca> Hi Inga, Sounds like your antibody needs heat retrieval to work on formalin fixed tissue. It really does not matter how you create the heat. Generally speaking, the lower the temperature, the longer the time in the heated buffer. You will have to experiment, but microwave should work fine. The problem with microwave oven is the possible hot spots, therefore giving you uneven staining. Always use the same amount of buffer, the same number of slides and rotating base, if you have one. Good luck! Katri Katri Tuomala St.Joseph's Healthcare Hamilton, Ontario, Canada ----- Original Message ----- From: "Inga Hansson" To: Sent: Thursday, October 30, 2003 9:09 AM Subject: [Histonet] pressure cooking > Hi all! > > I have an antibody that requires pressure cooking for 1 minute (after > having reached temperature) but I have no pressure cooker. Is it > possible to heat in microwave-owen instead and for how long? > > Inga > > -- > > Inga Hansson > Dept. neuroscience, > div. neurobiology > PO Box 587, BMC > Husargatan 3 > S-751 23 Uppsala > SWEDEN > > Phone: +46(18) 471 4384 > Fax: +46(18)559017 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From doscwk <@t> nus.edu.sg Thu Oct 30 19:33:01 2003 From: doscwk <@t> nus.edu.sg (Chan Wai Kam) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] RE: Microwave Tissue Processors Message-ID: Hi Loralee, My specimens are usually rabbit joints. I've no experience using a microwave for processing and need some advice. Can we use a normal microwave for this purpose? And what would be the optimum setting/temperature/time to use and processing time for each stage (fixation and decalcification)? Would really appreciate any advice from those who've been there and done that. Thanks in advance Julee Chan Orthopaedic Surgery National University of Singapore -----Original Message----- From: Gehan, Loralee [mailto:Loralee_Gehan@URMC.Rochester.edu] Sent: Thursday, October 30, 2003 9:09 PM To: Mary Reeves; Chan Wai Kam Cc: HistoNet Server Subject: RE: [Histonet] RE: Microwave Tissue Processors I work in an orthopaedics research lab as well and process many many many mouse hind limbs, calvaria, etc. We use a microwave processor from Hacker (milestone medical). It has decreased our time to process. We had some trouble at the start with some of our enzyme histochemistry. The trick to the machine is the heat. It was heating the samples up so much that one of our stains wasn't working. After much testing we figured out that all you have to do is decrease the temperature of the processing and increase the time. It was basically trial and error because the company markets these for rapid processing for surgical biopsies. We decal on the machine and we also do antigen retrieval and found that most of our antibodies worked well with it. We are still testing out the capabilities of the machine. But it has helped this lab become more efficient. Hope that helps. Loralee Gehan Orthopaedics Research Lab University of Rochester > ---------- > From: Chan Wai Kam > Sent: Wednesday, October 29, 2003 10:02 PM > To: Mary Reeves > Cc: HistoNet Server > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > Hi Mary, > > I was about to post a question to Histonet about the use of microwave > for processing of tissues when I came across your message below. I > hope to get some advice. > > I'm from Orthopaedics (research) and I process bone and cartilage > specimens the usual way through fixation in formalin, then > decalcification and so on. I'm just wondering whether I can use the > microwave to speed up the processing without affecting the quality of > the specimens. Our usual processing for bone takes around 3 weeks > until embedding so it would be great if we could have something that > can speed up the process without sacrifing quality. > > Would appreciate any advice out there. > > Thanks > Julee Chan > Orthopaedic Surgery > National University of Singapore > > > > From L.Driessen <@t> orthop.umcn.nl Fri Oct 31 01:33:39 2003 From: L.Driessen <@t> orthop.umcn.nl (Driessen, L.) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] RE: alkaline phosphatase stain on MMA sections Message-ID: <5B31B9F59E138A409E8F7C2183FF36F00DF490@umcnet13.umcn.nl> Hello Patty, I get pretty good result with this protocol. Alkaline phosphatase: BCIP-NBT method. (Manual of Histological Techniques; Bancroft/Cook) Controlmaterial: kidney, liver, spleen. Material: Frozen sections, paraffinsections, MMA-sections of (EDTA-decalcified) bone. - Remove the MMA and bring the slices through alcohol to water. - Pre-incubate EDTA-decalcified material one night with TRIS/MgCl2-buffer. - Incubate in alkaline phosphatase medium at 37?C (minimal time 30 minutes). Check microscopical. - Rinse with water. - Counterstain with Mayer haematoxylin, 1 minute. - Rinse with tapwater, 2 minutes. - Mount in watermounting medium (NBT disslove in in alcohol, xylene). Results: Alkaline phosphatase reactivity: Darkblue to black. Nuclei: Blue TRIS/ MgCl2-buffer: - 2,4 gram TRIS (hydroxymethylaminomethane). - 1 gram MgCl2. - 100 ml distilled water. Dissolve and bring pH to 9,0 with HCl. Veronal-buffer: - 1,9 gram Na-acetate (waterfree). - 2,9 gram Na-barbiturate (5,5 diethylbarbituurzuur, Na-salt) - 100 ml distilled water. Bring pH to 9,5 with 0,1-1N HCl. 1M MgCl2 : - 20 gram MgCl2 .6H2O. - 100 ml. distilled water. Alkaline phosphatase medium: (pH 9,5): - 12,5 milligram 5-Bromo-4-chloro-3-indolyl phosphate in 1 ml dimethylformamid. - 50 milliliter 0,2M Veronal-acetattebuffer, pH 9,5 - 25 milligram NBT (Nitrobluetetrazolium). - 0,4 milliliter 1M Magnesiumchlorid From lpwenk <@t> covad.net Fri Oct 31 04:57:16 2003 From: lpwenk <@t> covad.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Black pigment on Bronch Lavages References: <30C050525B881C4AAFF41E6D16543E68015D02E4@mail3.cls.ab.ca> Message-ID: <008a01c39f9d$c0076da0$8732fea9@hppav> I'm not certain that carbon birefringes when viewed with polarizing lenses. Formalin pigment will, as does silica and asbestos. But I don't think carbon does. Carbon will resist "bleaching" with potassium permanganate and sulfuric acid, while this will bleach out melanin. Try this: 2 slides of the patient tissue, on silane or poly-L-lysine, dried for a couple of hours at 60 degrees C. (This procedure tends to pull tissue off slides.) And 2 slides of melanin, also on the same type subbed slides and dried of same. Use another patient's case where there is carbon in lung tissue, prepare 2 slides of this the same way for your carbon control. Deparaffinize all 6 slides to water. Keep 1 pt. slide and 1 control slide in water. Place the other 1 pt. tissue and 1 each of the other controls (carbon lung and melanin) in a solution of 99.7 mL d. water + 0.3 mL sulfuric acid + 0.3 g potassium permanganate for about 1 hour at room temp. Gently rinse in d. water. Remove excess brown-colored potassium permanganate from tissue with 1% oxalic acid for 1-2 minutes. Gently rinse in d. water. Combine all 6 slides together. Stain with H&E. The melanin will be gone from the treated melanin control slides, but will still be there on the untreated control slides. The carbon control will remain positive in the treated and untreated. Then look at the patient's tissue - treated and untreated. That's all I can think of to do at a histology lab level. An EM scope with metal analysis would work, as would micro-incineration analysis. But those involve specialized labs. Let us know what happens. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: Kathy.Johnston@CLS.ab.ca To: histonet@pathology.swmed.edu Sent: Thursday, October 30, 2003 4:02 PM Subject: RE: [Histonet] Black pigment on Bronch Lavages As our pathologist was explaining his problem to me while we looked at the slide in particular under his microscope, he said that Carbon was not the item in question as it does not become refractile when he polarizes/darkfield his microscope (to be honest he was whizzing objectives and filters so fast I could barely follow him!). I know carbon is very common in lung, but to my eye as well, it is too fine and regular to resemble the stuff I usually see. -----Original Message----- From: thehud@ldd.net [mailto:thehud@ldd.net] Sent: Wednesday, October 29, 2003 3:31 PM To: Kathy.Johnston@CLS.ab.ca Subject: Re: [Histonet] Black pigment on Bronch Lavages dear kathy, how have you ruled out carbon, as this is so common. peter h. dohan, md -----Original Message----- From: Kathy.Johnston@CLS.ab.ca Sent: Oct 29, 2003 1:30 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Black pigment on Bronch Lavages One of our pathologists and myself have been trying to identify some black intracellular pigmentation in a bronch lavage. We have ruled out carbon, and bleaching the section did not work, therefore is not melanin. It is a very fine dark black pigment and appears quite uniform in shape and size. Our pathologist is thinking that it is lead (the patient is a long time professional painter), but lead stains are negative. My other thought is aluminum deposits but have not yet stained for this. I am hoping someone on the "Net" may have some idea of what this may be, and if there is a method for demonstrating it. Thanks very much in advance! Kathy Johnston Tech II - Special Stains Anatomic Pathology - FMC Calgary Laboratory Services 1403-29 Street NW Calgary AB, Canada T2N 2T9 403-944-4760 403-290-4093 fax kathy.johnston@cls.ab.ca -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031031/75237927/attachment.htm From gudrun.lang <@t> aon.at Fri Oct 31 05:02:20 2003 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] RE: Microwave Tissue Processors References: <95774A6A6036D411AFEA00D0B73C864308880471@exmc3.urmc.rochester.edu> Message-ID: <001601c39f9e$746cc380$0d12a8c0@SERVER> We do the same thing with our bone marrows like Loralee, but do not use any microwave decalcification afterwards. Our samples are at least four days in EDTA and than are processed in the VIP like all other biopsies. It works fine. Our tests with decalcification with mirowave instead of slowly EDTA were not satisfied. Gudrun Lang General Hospital Linz, Austria ----- Original Message ----- From: "Gehan, Loralee" To: ; "Gehan, Loralee" ; "'Mary Reeves'" Cc: ; "Katherine Raker" Sent: Thursday, October 30, 2003 9:30 PM Subject: RE: [Histonet] RE: Microwave Tissue Processors > We have the RHS-1 also. What we normally do for our decals is to let them > sit in EDTA for about a week sometimes longer depending on the amount of > things that we have to cut/process, etc. Then we run it on the decal cycle > in fresh EDTA at 37C for about 5 hours then process them the next day. We > have been getting beautiful results. > Do you do your entire processing schedule under vacuum? Or just the > paraffin? > > Loralee > > > ---------- > > From: Mary Reeves > > Sent: Thursday, October 30, 2003 11:13 AM > > To: doscwk@nus.edu.sg; Loralee_Gehan@urmc.rochester.edu > > Cc: histonet@lists.utsouthwestern.edu; Katherine Raker > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > Loralee, > > Which Hacker microwave do you have? We use the RHS-1 to process our > > decals. We are processing with vacuum, this allows us to process at lower > > temperatures and still have decreased processing times. We plan to start > > decalcification in the microwave soon. Any suggestions? > > > > Mary Reeves > > Technical Specialist > > Histology > > 1-352-265-0680 ext 7-2113 > > > > >>> "Gehan, Loralee" 10/30/03 08:08AM > > >>> > > I work in an orthopaedics research lab as well and process many many many > > mouse hind limbs, calvaria, etc. We use a microwave processor from Hacker > > (milestone medical). It has decreased our time to process. We had some > > trouble at the start with some of our enzyme histochemistry. > > The trick to the machine is the heat. It was heating the samples up so > > much > > that one of our stains wasn't working. After much testing we figured out > > that all you have to do is decrease the temperature of the processing and > > increase the time. It was basically trial and error because the company > > markets these for rapid processing for surgical biopsies. > > We decal on the machine and we also do antigen retrieval and found that > > most > > of our antibodies worked well with it. We are still testing out the > > capabilities of the machine. But it has helped this lab become more > > efficient. > > Hope that helps. > > > > Loralee Gehan > > Orthopaedics Research Lab > > University of Rochester > > > ---------- > > > From: Chan Wai Kam > > > Sent: Wednesday, October 29, 2003 10:02 PM > > > To: Mary Reeves > > > Cc: HistoNet Server > > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > > > Hi Mary, > > > > > > I was about to post a question to Histonet about the use of microwave > > > for processing of tissues when I came across your message below. I hope > > > to get some advice. > > > > > > I'm from Orthopaedics (research) and I process bone and cartilage > > > specimens the usual way through fixation in formalin, then > > > decalcification and so on. I'm just wondering whether I can use the > > > microwave to speed up the processing without affecting the quality of > > > the specimens. Our usual processing for bone takes around 3 weeks until > > > embedding so it would be great if we could have something that can speed > > > up the process without sacrifing quality. > > > > > > Would appreciate any advice out there. > > > > > > Thanks > > > Julee Chan > > > Orthopaedic Surgery > > > National University of Singapore > > > > > > > > > > > > -----Original Message----- > > > From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] > > > Sent: Wednesday, October 29, 2003 11:38 PM > > > To: ASelf@gmhsc.com > > > Cc: histonet@lists.utsouthwestern.edu; Katherine Raker > > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > > > > > > Amy, > > > Do you currently have a microwave tissue processor? We use the > > > Hacker/Milestone RHS-1 Microwave to process our fatty tissue. The times > > > that you see only represent the time at temperature, they do not include > > > the "ramp" up time (the time needed to bring the solution to the > > > appropriate processing temperature). > > > > > > Fixation - We fix the tissue cassettes overnight (6pm - 3am) in > > > Alcoholic Zinc Formalin Ethanol - 10 minutes at 65 C JFC - Produced by > > > Hacker. Isopropanol can be used instead, however, you may need to extend > > > the time at temperature. - 1 hour and 37 minutes at 68 C > > > Vaporization (Vacuum Drying) -Pressure at 500 mbar > > > Paraffin - 48 minutes at 65 C and pressure at 100 mbar > > > > > > This program takes approximately 4 hours to run, it replaces our 16 hour > > > overnight program that we previously used to process fatty tissue. The > > > only fatty tissue we do not process in the microwave is lipoma. > > > > > > Mary Reeves > > > Technical Specialist > > > Histology > > > 1-352-265-0680 ext 7-2113 > > > > > > >>> Amy Self 10/28/03 01:58PM >>> > > > > > > Mary, > > > > > > Could you share with me your procedure for processing fatty > > > tissue in the microwave. > > > > > > Thanks, > > > Amy Self > > > > > > -----Original Message----- > > > From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] > > > Sent: Tuesday, October 28, 2003 1:38 PM > > > To: histonet@lists.utsouthwestern.edu; thallada@noch.org > > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > > > > > > We microwave process all of our biopsy specimens and our fatty tissue > > > (except lipoma). > > > > > > Mary Reeves > > > Technical Specialist > > > Histology > > > 1-352-265-0680 ext 7-2113 > > > > > > >>> "Hallada, Teri" 10/28/03 11:42AM >>> > > > I was wondering if anyone out there is using a microwave tissue > > > processor for routine hospital tissues. Are there any regulations > > > applicable to instituting one, ie FDA approval? Teri Hallada BS MT CT > > > (ASCP) thallada@noch.org > > > > > > > > > > > > > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > Note: The information contained in this message may be privileged and > > > confidential and protected from disclosure. If the reader of this > > > message is not the intended recipient, or an employee or 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 > > > > > > _______________________________________________ > > > 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 Fri Oct 31 06:18:07 2003 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] Black pigment on Bronch Lavages Message-ID: Carbon is neither refractile nor birefringent. Nothing becomes refractile simply by looking at it through crossed polarizers. To appreciate refractility (better) you need to either close the sub-stage diaphragm or lower the condenser. BTW, asbestos fibres may be birefringent, but they are so small when dispersed in lung, that this is no way of detection, because they are simply too small to see with a light microscope, by any means. The asbestos bodies are of course, haemosiderin encrustations. 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: Kathy.Johnston@CLS.ab.ca [mailto:Kathy.Johnston@CLS.ab.ca] Sent: 30 October 2003 21:03 To: histonet@pathology.swmed.edu Subject: RE: [Histonet] Black pigment on Bronch Lavages As our pathologist was explaining his problem to me while we looked at the slide in particular under his microscope, he said that Carbon was not the item in question as it does not become refractile when he polarizes/darkfield his microscope (to be honest he was whizzing objectives and filters so fast I could barely follow him!). I know carbon is very common in lung, but to my eye as well, it is too fine and regular to resemble the stuff I usually see. -----Original Message----- From: thehud@ldd.net [mailto:thehud@ldd.net] Sent: Wednesday, October 29, 2003 3:31 PM To: Kathy.Johnston@CLS.ab.ca Subject: Re: [Histonet] Black pigment on Bronch Lavages dear kathy, how have you ruled out carbon, as this is so common. peter h. dohan, md -----Original Message----- From: Kathy.Johnston@CLS.ab.ca Sent: Oct 29, 2003 1:30 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Black pigment on Bronch Lavages One of our pathologists and myself have been trying to identify some black intracellular pigmentation in a bronch lavage. We have ruled out carbon, and bleaching the section did not work, therefore is not melanin. It is a very fine dark black pigment and appears quite uniform in shape and size. Our pathologist is thinking that it is lead (the patient is a long time professional painter), but lead stains are negative. My other thought is aluminum deposits but have not yet stained for this. I am hoping someone on the "Net" may have some idea of what this may be, and if there is a method for demonstrating it. Thanks very much in advance! Kathy Johnston Tech II - Special Stains Anatomic Pathology - FMC Calgary Laboratory Services 1403-29 Street NW Calgary AB, Canada T2N 2T9 403-944-4760 403-290-4093 fax kathy.johnston@cls.ab.ca -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031031/7d5bd049/attachment.htm From Terry.Marshall <@t> rothgen.nhs.uk Fri Oct 31 06:42:41 2003 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:07 2005 Subject: [Histonet] artifact quiz ANSWER Message-ID: Everyone has sent an answer to my email address rather than through histonet it seems (as was suggested by Herb at the website), so others will not know what has been suggested.. Residual water, paraffin and air bubbles seem to be the favoured suggestions. However the answer is bleaching by light, as I accidentally left my microscope on overnight with this area underneath. It was perfectly stained (by that I mean normal for here:-)) the day prior. The real clue, as alluded to by several was the perfect circle plus the fact that the H had survived to some extent, the E going walkabout, and therefore anything suggesting a barrier to staining is ruled out. Highly commended is: Could it be that someone had left the section on a fluorescence microscope with epi-illumination from a high power objective, causing photobleaching of the eosin in a circle that is visible at lower power? This commonly happens if you spend too long looking at one particular area by fluorescence microscopy. I suppose it could happen with ordinary illumination, given enough time. John Kiernan And, almost there was: Looks like an air bubble was trapped under the tissue and it popped sometime during the staining process. It reminds me also of an area on a fluorescent labelled section that has been quenched, but of course this is not fluorescence. Kathleen Spencer, HT ASCP Also almost there, except that the answer is the exact opposite, as it is light bleaching rather than light staining: If it is a complete circle of light staining (cannot tell from your picture) it is lack of deparaffinization. Ada T. Feldman, MS, HT/HTL(ASCP) BTW, have left microscope on before, but never seen this effect. Hope you all enjoyed. 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 From REEVEML <@t> shands.ufl.edu Fri Oct 31 07:19:06 2003 From: REEVEML <@t> shands.ufl.edu (Mary Reeves) Date: Fri Sep 16 15:22:08 2005 Subject: [Histonet] RE: Microwave Tissue Processors Message-ID: Just the paraffin Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 >>> "Gehan, Loralee" 10/30/03 03:30PM >>> We have the RHS-1 also. What we normally do for our decals is to let them sit in EDTA for about a week sometimes longer depending on the amount of things that we have to cut/process, etc. Then we run it on the decal cycle in fresh EDTA at 37C for about 5 hours then process them the next day. We have been getting beautiful results. Do you do your entire processing schedule under vacuum? Or just the paraffin? Loralee > ---------- > From: Mary Reeves > Sent: Thursday, October 30, 2003 11:13 AM > To: doscwk@nus.edu.sg; Loralee_Gehan@urmc.rochester.edu > Cc: histonet@lists.utsouthwestern.edu; Katherine Raker > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > Loralee, > Which Hacker microwave do you have? We use the RHS-1 to process our > decals. We are processing with vacuum, this allows us to process at lower > temperatures and still have decreased processing times. We plan to start > decalcification in the microwave soon. Any suggestions? > > Mary Reeves > Technical Specialist > Histology > 1-352-265-0680 ext 7-2113 > > >>> "Gehan, Loralee" 10/30/03 08:08AM > >>> > I work in an orthopaedics research lab as well and process many many many > mouse hind limbs, calvaria, etc. We use a microwave processor from Hacker > (milestone medical). It has decreased our time to process. We had some > trouble at the start with some of our enzyme histochemistry. > The trick to the machine is the heat. It was heating the samples up so > much > that one of our stains wasn't working. After much testing we figured out > that all you have to do is decrease the temperature of the processing and > increase the time. It was basically trial and error because the company > markets these for rapid processing for surgical biopsies. > We decal on the machine and we also do antigen retrieval and found that > most > of our antibodies worked well with it. We are still testing out the > capabilities of the machine. But it has helped this lab become more > efficient. > Hope that helps. > > Loralee Gehan > Orthopaedics Research Lab > University of Rochester > > ---------- > > From: Chan Wai Kam > > Sent: Wednesday, October 29, 2003 10:02 PM > > To: Mary Reeves > > Cc: HistoNet Server > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > Hi Mary, > > > > I was about to post a question to Histonet about the use of microwave > > for processing of tissues when I came across your message below. I hope > > to get some advice. > > > > I'm from Orthopaedics (research) and I process bone and cartilage > > specimens the usual way through fixation in formalin, then > > decalcification and so on. I'm just wondering whether I can use the > > microwave to speed up the processing without affecting the quality of > > the specimens. Our usual processing for bone takes around 3 weeks until > > embedding so it would be great if we could have something that can speed > > up the process without sacrifing quality. > > > > Would appreciate any advice out there. > > > > Thanks > > Julee Chan > > Orthopaedic Surgery > > National University of Singapore > > > > > > > > -----Original Message----- > > From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] > > Sent: Wednesday, October 29, 2003 11:38 PM > > To: ASelf@gmhsc.com > > Cc: histonet@lists.utsouthwestern.edu; Katherine Raker > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > > > Amy, > > Do you currently have a microwave tissue processor? We use the > > Hacker/Milestone RHS-1 Microwave to process our fatty tissue. The times > > that you see only represent the time at temperature, they do not include > > the "ramp" up time (the time needed to bring the solution to the > > appropriate processing temperature). > > > > Fixation - We fix the tissue cassettes overnight (6pm - 3am) in > > Alcoholic Zinc Formalin Ethanol - 10 minutes at 65 C JFC - Produced by > > Hacker. Isopropanol can be used instead, however, you may need to extend > > the time at temperature. - 1 hour and 37 minutes at 68 C > > Vaporization (Vacuum Drying) -Pressure at 500 mbar > > Paraffin - 48 minutes at 65 C and pressure at 100 mbar > > > > This program takes approximately 4 hours to run, it replaces our 16 hour > > overnight program that we previously used to process fatty tissue. The > > only fatty tissue we do not process in the microwave is lipoma. > > > > Mary Reeves > > Technical Specialist > > Histology > > 1-352-265-0680 ext 7-2113 > > > > >>> Amy Self 10/28/03 01:58PM >>> > > > > Mary, > > > > Could you share with me your procedure for processing fatty > > tissue in the microwave. > > > > Thanks, > > Amy Self > > > > -----Original Message----- > > From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] > > Sent: Tuesday, October 28, 2003 1:38 PM > > To: histonet@lists.utsouthwestern.edu; thallada@noch.org > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > > > We microwave process all of our biopsy specimens and our fatty tissue > > (except lipoma). > > > > Mary Reeves > > Technical Specialist > > Histology > > 1-352-265-0680 ext 7-2113 > > > > >>> "Hallada, Teri" 10/28/03 11:42AM >>> > > I was wondering if anyone out there is using a microwave tissue > > processor for routine hospital tissues. Are there any regulations > > applicable to instituting one, ie FDA approval? Teri Hallada BS MT CT > > (ASCP) thallada@noch.org > > > > > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > Note: The information contained in this message may be privileged and > > confidential and protected from disclosure. If the reader of this > > message is not the intended recipient, or an employee or 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 > > > > _______________________________________________ > > 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 REEVEML <@t> shands.ufl.edu Fri Oct 31 08:03:12 2003 From: REEVEML <@t> shands.ufl.edu (Mary Reeves) Date: Fri Sep 16 15:22:08 2005 Subject: [Histonet] RE: Microwave Tissue Processors Message-ID: When you tried decalcification in the microwave did you use EDTA? At what temperature? Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 >>> "Gudrun Lang" 10/31/03 06:02AM >>> We do the same thing with our bone marrows like Loralee, but do not use any microwave decalcification afterwards. Our samples are at least four days in EDTA and than are processed in the VIP like all other biopsies. It works fine. Our tests with decalcification with mirowave instead of slowly EDTA were not satisfied. Gudrun Lang General Hospital Linz, Austria ----- Original Message ----- From: "Gehan, Loralee" To: ; "Gehan, Loralee" ; "'Mary Reeves'" Cc: ; "Katherine Raker" Sent: Thursday, October 30, 2003 9:30 PM Subject: RE: [Histonet] RE: Microwave Tissue Processors > We have the RHS-1 also. What we normally do for our decals is to let them > sit in EDTA for about a week sometimes longer depending on the amount of > things that we have to cut/process, etc. Then we run it on the decal cycle > in fresh EDTA at 37C for about 5 hours then process them the next day. We > have been getting beautiful results. > Do you do your entire processing schedule under vacuum? Or just the > paraffin? > > Loralee > > > ---------- > > From: Mary Reeves > > Sent: Thursday, October 30, 2003 11:13 AM > > To: doscwk@nus.edu.sg; Loralee_Gehan@urmc.rochester.edu > > Cc: histonet@lists.utsouthwestern.edu; Katherine Raker > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > Loralee, > > Which Hacker microwave do you have? We use the RHS-1 to process our > > decals. We are processing with vacuum, this allows us to process at lower > > temperatures and still have decreased processing times. We plan to start > > decalcification in the microwave soon. Any suggestions? > > > > Mary Reeves > > Technical Specialist > > Histology > > 1-352-265-0680 ext 7-2113 > > > > >>> "Gehan, Loralee" 10/30/03 08:08AM > > >>> > > I work in an orthopaedics research lab as well and process many many many > > mouse hind limbs, calvaria, etc. We use a microwave processor from Hacker > > (milestone medical). It has decreased our time to process. We had some > > trouble at the start with some of our enzyme histochemistry. > > The trick to the machine is the heat. It was heating the samples up so > > much > > that one of our stains wasn't working. After much testing we figured out > > that all you have to do is decrease the temperature of the processing and > > increase the time. It was basically trial and error because the company > > markets these for rapid processing for surgical biopsies. > > We decal on the machine and we also do antigen retrieval and found that > > most > > of our antibodies worked well with it. We are still testing out the > > capabilities of the machine. But it has helped this lab become more > > efficient. > > Hope that helps. > > > > Loralee Gehan > > Orthopaedics Research Lab > > University of Rochester > > > ---------- > > > From: Chan Wai Kam > > > Sent: Wednesday, October 29, 2003 10:02 PM > > > To: Mary Reeves > > > Cc: HistoNet Server > > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > > > Hi Mary, > > > > > > I was about to post a question to Histonet about the use of microwave > > > for processing of tissues when I came across your message below. I hope > > > to get some advice. > > > > > > I'm from Orthopaedics (research) and I process bone and cartilage > > > specimens the usual way through fixation in formalin, then > > > decalcification and so on. I'm just wondering whether I can use the > > > microwave to speed up the processing without affecting the quality of > > > the specimens. Our usual processing for bone takes around 3 weeks until > > > embedding so it would be great if we could have something that can speed > > > up the process without sacrifing quality. > > > > > > Would appreciate any advice out there. > > > > > > Thanks > > > Julee Chan > > > Orthopaedic Surgery > > > National University of Singapore > > > > > > > > > > > > -----Original Message----- > > > From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] > > > Sent: Wednesday, October 29, 2003 11:38 PM > > > To: ASelf@gmhsc.com > > > Cc: histonet@lists.utsouthwestern.edu; Katherine Raker > > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > > > > > > Amy, > > > Do you currently have a microwave tissue processor? We use the > > > Hacker/Milestone RHS-1 Microwave to process our fatty tissue. The times > > > that you see only represent the time at temperature, they do not include > > > the "ramp" up time (the time needed to bring the solution to the > > > appropriate processing temperature). > > > > > > Fixation - We fix the tissue cassettes overnight (6pm - 3am) in > > > Alcoholic Zinc Formalin Ethanol - 10 minutes at 65 C JFC - Produced by > > > Hacker. Isopropanol can be used instead, however, you may need to extend > > > the time at temperature. - 1 hour and 37 minutes at 68 C > > > Vaporization (Vacuum Drying) -Pressure at 500 mbar > > > Paraffin - 48 minutes at 65 C and pressure at 100 mbar > > > > > > This program takes approximately 4 hours to run, it replaces our 16 hour > > > overnight program that we previously used to process fatty tissue. The > > > only fatty tissue we do not process in the microwave is lipoma. > > > > > > Mary Reeves > > > Technical Specialist > > > Histology > > > 1-352-265-0680 ext 7-2113 > > > > > > >>> Amy Self 10/28/03 01:58PM >>> > > > > > > Mary, > > > > > > Could you share with me your procedure for processing fatty > > > tissue in the microwave. > > > > > > Thanks, > > > Amy Self > > > > > > -----Original Message----- > > > From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] > > > Sent: Tuesday, October 28, 2003 1:38 PM > > > To: histonet@lists.utsouthwestern.edu; thallada@noch.org > > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > > > > > > We microwave process all of our biopsy specimens and our fatty tissue > > > (except lipoma). > > > > > > Mary Reeves > > > Technical Specialist > > > Histology > > > 1-352-265-0680 ext 7-2113 > > > > > > >>> "Hallada, Teri" 10/28/03 11:42AM >>> > > > I was wondering if anyone out there is using a microwave tissue > > > processor for routine hospital tissues. Are there any regulations > > > applicable to instituting one, ie FDA approval? Teri Hallada BS MT CT > > > (ASCP) thallada@noch.org > > > > > > > > > > > > > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > Note: The information contained in this message may be privileged and > > > confidential and protected from disclosure. If the reader of this > > > message is not the intended recipient, or an employee or 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 > > > > > > _______________________________________________ > > > 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 Fri Oct 31 08:33:56 2003 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:22:08 2005 Subject: [Histonet] Dionized vs distilled water Message-ID: <494304423C63E246A5CF87A3AEEB577011B5A4@bumail01.barrynet.barry.edu> Tap water varies a little from one water system to another. It is usually a very dilute solution of the carbonates and chlorides of calcium, sodium, and copper. Its pH is usually a little over 7, so it acts as a very weak base. (Mine is 7.4) It must be used when the protocol calls for it. Distilled and deionized water contain so few ions that neither one will act as a weak base. For most purposes, distilled and deionized water are interchangeable. It is easier to contaminate a deionizing system, but one can contaminate a distilled water system. Small, lab-sized systems are less likely to become contaminated with bacteria or minerals than large building-sized systems are. If you need really pure water, boil 600 ml of deionized water and add 6 mg of potassium permanganate to it and distill the water. Discard the first 100 ml of distillate. The next 300 ml will be very pure water, but I can think of no biological use for it. (Such very pure water is sometimes used for physics experiments.) Both distilled and deionized water pick up carbon dioxide on standing and become weak solutions of carbonic acid. If you work alone, its pH will probably be 6.9; if there are several people in the lab, the pH may fall to 6.8. -----Original Message----- From: Morken, Tim - Labvision [mailto:tpmorken@labvision.com] Sent: Tuesday, October 28, 2003 12:53 PM To: 'JCarpenter764@aol.com'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Dionized vs distilled water Distilled water is classically produced by heating water to evaporation and subsequent condensing on a cold surface. In the process most impurities are either evaporated off ahead of the water (in the case of most organics), or left behind (in the case of minerals). The water is also effectively deionized because the salts are left behind. It is fairly pure water. To get very pure water it needs to be re-distilled several times. Deionized water is classically passed through a salt bed or ionized resin bed that captures the mineral ions (ie, a "water softener"). The water is not necessarily pure, however, especially in regards to organic chemicals. Reverse osmosis is also used now days to deionize water. High quality water systems these days are some combination of filters, distillation, deionizing resins and reverse osmosis. Tim Morken -----Original Message----- From: JCarpenter764@aol.com [mailto:JCarpenter764@aol.com] Sent: Tuesday, October 28, 2003 7:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) while studying for my exam on the different fixatives and there ingredients....i have noticed that some call for distilled water and some use the term deionized water. Is there a difference? 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) -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031031/34adf8d3/attachment.htm From gudrun.lang <@t> aon.at Fri Oct 31 09:59:31 2003 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:08 2005 Subject: **SPAM** Re: [Histonet] RE: Microwave Tissue Processors References: Message-ID: <001601c39fc7$f8a6dfe0$0d12a8c0@SERVER> Mary, We took the well fixed bone marrow probes und tried decalcification with EDTA I think at 37 to 40 degrees for a few ours. Actually it was about two years ago and I only remember the bad result. We have about six to ten bone marrow probes over a week and stain them once a week. Therefore we histotechs believe, that the number is too small for the big effort. On the other hand we did not like the noise and the small display of the machine. best wishes Gudrun Lang general hospital Linz, Austria ----- Original Message ----- From: "Mary Reeves" To: ; ; Sent: Friday, October 31, 2003 3:03 PM Subject: **SPAM** Re: [Histonet] RE: Microwave Tissue Processors When you tried decalcification in the microwave did you use EDTA? At what temperature? Mary Reeves Technical Specialist Histology 1-352-265-0680 ext 7-2113 >>> "Gudrun Lang" 10/31/03 06:02AM >>> We do the same thing with our bone marrows like Loralee, but do not use any microwave decalcification afterwards. Our samples are at least four days in EDTA and than are processed in the VIP like all other biopsies. It works fine. Our tests with decalcification with microwave instead of slowly EDTA were not satisfying. Gudrun Lang General Hospital Linz, Austria ----- Original Message ----- From: "Gehan, Loralee" To: ; "Gehan, Loralee" ; "'Mary Reeves'" Cc: ; "Katherine Raker" Sent: Thursday, October 30, 2003 9:30 PM Subject: RE: [Histonet] RE: Microwave Tissue Processors > We have the RHS-1 also. What we normally do for our decals is to let them > sit in EDTA for about a week sometimes longer depending on the amount of > things that we have to cut/process, etc. Then we run it on the decal cycle > in fresh EDTA at 37C for about 5 hours then process them the next day. We > have been getting beautiful results. > Do you do your entire processing schedule under vacuum? Or just the > paraffin? > > Loralee > > > ---------- > > From: Mary Reeves > > Sent: Thursday, October 30, 2003 11:13 AM > > To: doscwk@nus.edu.sg; Loralee_Gehan@urmc.rochester.edu > > Cc: histonet@lists.utsouthwestern.edu; Katherine Raker > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > Loralee, > > Which Hacker microwave do you have? We use the RHS-1 to process our > > decals. We are processing with vacuum, this allows us to process at lower > > temperatures and still have decreased processing times. We plan to start > > decalcification in the microwave soon. Any suggestions? > > > > Mary Reeves > > Technical Specialist > > Histology > > 1-352-265-0680 ext 7-2113 > > > > >>> "Gehan, Loralee" 10/30/03 08:08AM > > >>> > > I work in an orthopaedics research lab as well and process many many many > > mouse hind limbs, calvaria, etc. We use a microwave processor from Hacker > > (milestone medical). It has decreased our time to process. We had some > > trouble at the start with some of our enzyme histochemistry. > > The trick to the machine is the heat. It was heating the samples up so > > much > > that one of our stains wasn't working. After much testing we figured out > > that all you have to do is decrease the temperature of the processing and > > increase the time. It was basically trial and error because the company > > markets these for rapid processing for surgical biopsies. > > We decal on the machine and we also do antigen retrieval and found that > > most > > of our antibodies worked well with it. We are still testing out the > > capabilities of the machine. But it has helped this lab become more > > efficient. > > Hope that helps. > > > > Loralee Gehan > > Orthopaedics Research Lab > > University of Rochester > > > ---------- > > > From: Chan Wai Kam > > > Sent: Wednesday, October 29, 2003 10:02 PM > > > To: Mary Reeves > > > Cc: HistoNet Server > > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > > > Hi Mary, > > > > > > I was about to post a question to Histonet about the use of microwave > > > for processing of tissues when I came across your message below. I hope > > > to get some advice. > > > > > > I'm from Orthopaedics (research) and I process bone and cartilage > > > specimens the usual way through fixation in formalin, then > > > decalcification and so on. I'm just wondering whether I can use the > > > microwave to speed up the processing without affecting the quality of > > > the specimens. Our usual processing for bone takes around 3 weeks until > > > embedding so it would be great if we could have something that can speed > > > up the process without sacrifing quality. > > > > > > Would appreciate any advice out there. > > > > > > Thanks > > > Julee Chan > > > Orthopaedic Surgery > > > National University of Singapore > > > > > > > > > > > > -----Original Message----- > > > From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] > > > Sent: Wednesday, October 29, 2003 11:38 PM > > > To: ASelf@gmhsc.com > > > Cc: histonet@lists.utsouthwestern.edu; Katherine Raker > > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > > > > > > Amy, > > > Do you currently have a microwave tissue processor? We use the > > > Hacker/Milestone RHS-1 Microwave to process our fatty tissue. The times > > > that you see only represent the time at temperature, they do not include > > > the "ramp" up time (the time needed to bring the solution to the > > > appropriate processing temperature). > > > > > > Fixation - We fix the tissue cassettes overnight (6pm - 3am) in > > > Alcoholic Zinc Formalin Ethanol - 10 minutes at 65 C JFC - Produced by > > > Hacker. Isopropanol can be used instead, however, you may need to extend > > > the time at temperature. - 1 hour and 37 minutes at 68 C > > > Vaporization (Vacuum Drying) -Pressure at 500 mbar > > > Paraffin - 48 minutes at 65 C and pressure at 100 mbar > > > > > > This program takes approximately 4 hours to run, it replaces our 16 hour > > > overnight program that we previously used to process fatty tissue. The > > > only fatty tissue we do not process in the microwave is lipoma. > > > > > > Mary Reeves > > > Technical Specialist > > > Histology > > > 1-352-265-0680 ext 7-2113 > > > > > > >>> Amy Self 10/28/03 01:58PM >>> > > > > > > Mary, > > > > > > Could you share with me your procedure for processing fatty > > > tissue in the microwave. > > > > > > Thanks, > > > Amy Self > > > > > > -----Original Message----- > > > From: Mary Reeves [mailto:REEVEML@shands.ufl.edu] > > > Sent: Tuesday, October 28, 2003 1:38 PM > > > To: histonet@lists.utsouthwestern.edu; thallada@noch.org > > > Subject: RE: [Histonet] RE: Microwave Tissue Processors > > > > > > > > > We microwave process all of our biopsy specimens and our fatty tissue > > > (except lipoma). > > > > > > Mary Reeves > > > Technical Specialist > > > Histology > > > 1-352-265-0680 ext 7-2113 > > > > > > >>> "Hallada, Teri" 10/28/03 11:42AM >>> > > > I was wondering if anyone out there is using a microwave tissue > > > processor for routine hospital tissues. Are there any regulations > > > applicable to instituting one, ie FDA approval? Teri Hallada BS MT CT > > > (ASCP) thallada@noch.org > > > > > > > > > > > > > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > Note: The information contained in this message may be privileged and > > > confidential and protected from disclosure. If the reader of this > > > message is not the intended recipient, or an employee or 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 > > > > > > _______________________________________________ > > > 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 ttroyer <@t> pcllab.com Fri Oct 31 10:29:09 2003 From: ttroyer <@t> pcllab.com (Travis Troyer) Date: Fri Sep 16 15:22:08 2005 Subject: [Histonet] AMACR Message-ID: <000a01c39fcc$1cca4a70$6601010a@Peterson.local> Our pathologist here was interested in the following antibody, alpha-methacryl-CoA racemase. I was wondering in anyone uses this antibody or knows of a vendor for the antibody. Thanks, Travis Troyer Peterson Clinical Lab Manhattan, KS -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031031/7993c3c2/attachment.htm From sa.drew <@t> hosp.wisc.edu Fri Oct 31 10:42:09 2003 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:22:08 2005 Subject: [Histonet] AMACR Message-ID: I recently worked up BioCare Medical's antibody P504S(AMACR). Our director anticipates it being ordered as a dual stain w/ high molecular weight cytokeratin. We're still working on the dual stain. Let me know if you want more particulars. Sally Ann Drew-MT(ASCP) Univ. of Wisconsin Hospital & Clinics IHC/ISH Laboratory 600 Highland Ave. VAH-DM223 Madison, WI 53792-2472 608-265-6596 sa.drew@hosp.wisc.edu -----Original Message----- From: Travis Troyer [mailto:ttroyer@pcllab.com] Sent: Friday, October 31, 2003 10:29 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] AMACR Our pathologist here was interested in the following antibody, alpha-methacryl-CoA racemase. I was wondering in anyone uses this antibody or knows of a vendor for the antibody. Thanks, Travis Troyer Peterson Clinical Lab Manhattan, KS -------------- next part -------------- An HTML attachment was scrubbed... URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20031031/8369b13d/attachment.htm From Karen.Dresser <@t> umassmed.edu Fri Oct 31 11:02:48 2003 From: Karen.Dresser <@t> umassmed.edu (Dresser, Karen) Date: Fri Sep 16 15:22:08 2005 Subject: [Histonet] AMACR Message-ID: <5B238BE7511C7D4FB736CF0451643D530F7646@edunivmail03.ad.umassmed.edu> The dual stain works great! I've used P504 for quite some time, and recently worked up the dual stain. For ours, we use HMW-CK and p63 together (HRP-DAB) with p504 (AP-Fast Red). Two important things about working with p504: 1) make sure your control has prostate CA, otherwise you may not get staining. 2) make sure your slides are not old -- you'll get best results from slides under 1 month old. Enjoy! Karen Dresser Research Histotechnician UMass Medical School -----Original Message----- From: Drew Sally A. [mailto:sa.drew@hosp.wisc.edu] Sent: Friday, October 31, 2003 11:42 AM To: Histonet Subject: RE: [Histonet] AMACR I recently worked up BioCare Medical's antibody P504S(AMACR). Our director anticipates it being ordered as a dual stain w/ high molecular weight cytokeratin. We're still working on the dual stain. Let me know if you want more particulars. Sally Ann Drew-MT(ASCP) Univ. of Wisconsin Hospital & Clinics IHC/ISH Laboratory 600 Highland Ave. VAH-DM223 Madison, WI 53792-2472 608-265-6596 sa.drew@hosp.wisc.edu -----Original Message----- From: Travis Troyer [mailto:ttroyer@pcllab.com] Sent: Friday, October 31, 2003 10:29 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] AMACR Our pathologist here was interested in the following antibody, alpha-methacryl-CoA racemase. I was wondering in anyone uses this antibody or knows of a vendor for the antibody. Thanks, Travis Troyer Peterson Clinical Lab Manhattan, KS From pdelvent <@t> wyoming.com Fri Oct 31 11:06:35 2003 From: pdelvent <@t> wyoming.com (Priscilla Delventhal) Date: Fri Sep 16 15:22:08 2005 Subject: [Histonet] Histology books for sale Message-ID: Hello Histonetters - I am semi-retired now and would like to sell some of the books and periodicals I've accumulated over the years. Most of the periodicals are free for the postage and the books range from $1.00 to $5.00. Most of these are from the 60's and 70's when I was actively teaching. Some are medical books for pathologists, but I found them helpful. Even though they are not recent, they are still very helpful to histology students and for $5.00 it would be pretty hard to find this material elsewhere. A couple of the books are from the 60's and are possibly considered classics. I will send them book rate and require a money order in hand before they will be sent. I will be on temporary assignment from October 15th through December 10th so you need to contact me ASAP. Contact me by e-mail and I will send you the 5 page list. Thanks - Priscilla Delventhal in Central Wyoming From t-kuzniar <@t> md.northwestern.edu Fri Oct 31 11:30:55 2003 From: t-kuzniar <@t> md.northwestern.edu (t-kuzniar@md.northwestern.edu) Date: Fri Sep 16 15:22:08 2005 Subject: [Histonet] b-galactosidase in lung Message-ID: <200310311731.h9VHV9Ej018619@casbah.it.northwestern.edu> Hi, I'm having troubles finding my b-gal-expressing plasmid in the lung tissue. I do 5-10 um frozen sections of murine lungs, fix them for 10 minutes in 0.2% glutaraldehyde, wash twice in a detergent wash, and incubate overnight in regular lacZ staining solution. I am sure my lacZ is there, because i can see it when I stain my lungs macroscopically. Thanks Tom Kuzniar From t-kuzniar <@t> md.northwestern.edu Fri Oct 31 13:08:38 2003 From: t-kuzniar <@t> md.northwestern.edu (t-kuzniar@md.northwestern.edu) Date: Fri Sep 16 15:22:08 2005 Subject: [Histonet] Once more b-galactosidase in lung Message-ID: <200310311908.h9VJ8nxZ006137@casbah.it.northwestern.edu> Thanks for the advice, one more question. B-galactosidase is heat- and light-sensitive. How sensitive is it? I'm cutting my frozen sections without any shielding and when they're drying out for 10-30 minutes, they're unprotected from light either. I initially did not think it would matter, but maybe? Thanks Tom From jkiernan <@t> uwo.ca Fri Oct 31 10:58:22 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:22:08 2005 Subject: [Histonet] Sirius red References: <4.3.2.7.2.20031030145119.00ccb050@algranth.inbox.email.arizona.edu> Message-ID: <3FA294AE.40B5E8A9@uwo.ca> CI 35780 is sirius red F3B (= Direct red 80). It should work in a picro-sirius red method. If it really fails, please let us all know the details - supplier, batch etc. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ______________________________ Andrea Grantham wrote: > > I have a quick question about Sirius red since it is a recent topic of > discussion on histonet. > I was asked to do the stain and given a protocol calling for Sirius red F3B > (CI 35782). I have Sirius red CI 35780. They said that they have tried the > stain using CI 35780 and it didn't work but they cannot find the other > Sirius red with a CI# 35782 in any of the catalogs. We have tried Sigma, > Polysciences, PolyScientific, Newcomer, and some others... > Any suggestions where I can find it? > Andi Grantham > ..................................................................... > : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > : Sr. Research Specialist University of Arizona : > : (office: AHSC 4212) P.O. Box 245044 : > : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : > :...................................................................: > http://www.cba.arizona.edu/histology-lab.html > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Oct 31 11:52:13 2003 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:08 2005 Subject: [Histonet] PBS Message-ID: <3.0.6.32.20031031105213.00bc7760@gemini.msu.montana.edu> Trisha, To make your life easier, buy Dulbeccos PBS (without added Mg) from Sigma #5652 a jar of this will make up 50 liters. You weigh out 10g and dissolve in 1 liter of distilled or deionized water. The pH is always right on at 7.4. It saves so much time to buy preweighed PBS (others also available from any reliable company, probably even Fisher, VWR, etc) in little packets to make up a liter. TRIS buffered saline is also available, and often in the pH range you need. Just go shopping in Sigma catalog. We had a horrible experience with an in house PBS preparation, supposedly guaranteed by researcher to be "perfect" PBS, but his technician had made it up incorrectly and blew away my sections. We now buy DPBS from Sigma and never have problems plus saving time of having to make buffer from scratch. Unfortunately, this experience makes me very suspect of other peoples preparations - would rather do it myself the easy way. Good luck Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From jkiernan <@t> uwo.ca Fri Oct 31 12:51:54 2003 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:22:08 2005 Subject: [Histonet] b-galactosidase in lung References: <200310311731.h9VHV9Ej018619@casbah.it.northwestern.edu> Message-ID: <3FA2AF4A.FB23D1C1@uwo.ca> According to Takahashi et al J Histochem Cytochem 51(4):553-554 (2003) beta galactosidase staining was strongest after fixation of cryosections for 10 min to 8 hrs at room temperature in 100% acetone. There was weaker staining after 0.2% glutaraldehyde and almost no staining after 4% formaldehyde (from paraformaldehyde) - all done for the same times & temperature. They were looking at muscles from LacZ transgenic rats, and they used the usual indigogenic method, with a 30 minute incubation. Could your detergent wash be doing something nasty? It seems a very harsh thing to do to sections of almost unfixed tissue, and there's no obvious reason for it. -- ------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan@uwo.ca http://publish.uwo.ca/~jkiernan/ ______________________________________ t-kuzniar@md.northwestern.edu wrote: > > Hi, > I'm having troubles finding my b-gal-expressing plasmid in the lung > tissue. I do 5-10 um frozen sections of murine lungs, fix them for 10 > minutes in 0.2% glutaraldehyde, wash twice in a detergent wash, and > incubate overnight in regular lacZ staining solution. > > I am sure my lacZ is there, because i can see it when I stain my lungs > macroscopically. > > Thanks > > Tom Kuzniar > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RITA.ANGEL <@t> UC.EDU Fri Oct 31 13:23:17 2003 From: RITA.ANGEL <@t> UC.EDU (Rita Angel) Date: Fri Sep 16 15:22:08 2005 Subject: [Histonet] collagen I staining Message-ID: <5.1.0.14.2.20031031141929.00b3edc8@ucmail3.uc.edu> Hello Histonetters, Someone in our lab has been trying to stain for collagen I antibody in rabbit tissue. She's tried the antibody from several different companies and has used different antigen retrievals, but has been unable to get any positive staining. The only staining she's gotten is background. These are paraffin sections. The companies she bought the antibodies from has not been helpful. She's stained with Collagen type II & III and had no trouble with these. Any suggestions would be helpful. Thank you, Rita Angel, HT University of Cincinnati From dmikita <@t> wmcnet.org Fri Oct 31 13:40:38 2003 From: dmikita <@t> wmcnet.org (Daryl Mikita) Date: Fri Sep 16 15:22:08 2005 Subject: [Histonet] Histology Position Open Message-ID: Hello Everyone, Just letting you know that we are looking for a new full-time histotech at the Wyoming Medical Center. I don't know any of information, but you can look at the hospitals web page: http://www.wmcnet.org or you can call human resource at 1-307-577-2406 TTYL Daryl Mikita, HT(ASCP) Wyoming Medical Center 1233 E. 2nd St. Casper, WY 82601 From jstaruk <@t> masshistology.com Fri Oct 31 13:50:30 2003 From: jstaruk <@t> masshistology.com (Mass Histology Service) Date: Fri Sep 16 15:22:08 2005 Subject: [Histonet] collagen I staining In-Reply-To: <5.1.0.14.2.20031031141929.00b3edc8@ucmail3.uc.edu> Message-ID: We've had good luck with Calbiochem's anti-collagen I (Cat. # CP17L), 1:50 following pronase pre-treatment on rabbit bones. Jim ____________________ James Staruk, HT(ASCP) Mass Histology Service A division of DVMLab, inc. www.masshistology.com -----Original Message----- From: histonet-admin@lists.utsouthwestern.edu [mailto:histonet-admin@lists.utsouthwestern.edu]On Behalf Of Rita Angel Sent: Friday, October 31, 2003 2:23 PM To: histonet@pathology.swmed.edu Subject: [Histonet] collagen I staining Hello Histonetters, Someone in our lab has been trying to stain for collagen I antibody in rabbit tissue. She's tried the antibody from several different companies and has used different antigen retrievals, but has been unable to get any positive staining. The only staining she's gotten is background. These are paraffin sections. The companies she bought the antibodies from has not been helpful. She's stained with Collagen type II & III and had no trouble with these. Any suggestions would be helpful. Thank you, Rita Angel, HT University of Cincinnati _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> msn.com Fri Oct 31 13:57:23 2003 From: pruegg <@t> msn.com (Patsy Ruegg) Date: Fri Sep 16 15:22:08 2005 Subject: [Histonet] Sirius red Message-ID: Andi, When I was at the U I got Sirius Red thus: It is made by Chroma Gsellschaft Schmid GmBH + Co. D-7316 Konegen/N West Germany Distributed in the US by Roboz Surgical Instrument Co. Inc. 100 Connecticut Avenue N.W. Washington, D.C. 20036 Haven't bought it in years. I did just recently used a Picro-Sirius Red cat.# s2365 from Polyscientific, not sure of the C.I. # but it worked just like the stuff I had used before from Germany. Did you try the one's from the other places or did you just think that it would not work per your investigator? Perhaps they did not mix it with saturated picric acid, you need that for it to work properly. I do an alcian blue counter stain before the picro-sirius stain and it is beautiful. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 HP-303-644-4538 HF-303-644-3377 hm email pruegg@msn.com wk email pruegg@colobio.com 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. ----Original Message Follows---- From: Andrea Grantham To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Sirius red Date: Thu, 30 Oct 2003 15:08:08 -0700 I have a quick question about Sirius red since it is a recent topic of discussion on histonet. I was asked to do the stain and given a protocol calling for Sirius red F3B (CI 35782). I have Sirius red CI 35780. They said that they have tried the stain using CI 35780 and it didn't work but they cannot find the other Sirius red with a CI# 35782 in any of the catalogs. We have tried Sigma, Polysciences, PolyScientific, Newcomer, and some others... Any suggestions where I can find it? Andi Grantham ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Enjoy MSN 8 patented spam control and more with MSN 8 Dial-up Internet Service. Try it FREE for one month! http://join.msn.com/?page=dept/dialup